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出版者・発行元：Zoological Society of Japan  

DOI： 10.2108/zs200038

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出版者・発行元：Elsevier BV  

DOI： 10.1016/j.neuroscience.2020.04.047

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出版者・発行元：Elsevier BV  

DOI： 10.1016/j.cbpa.2020.110700

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記述言語：英語  

Thirst has evolved for vertebrate terrestrial adaptation. We previously showed that buccal drying induced a series of drinking behaviours (migration to water-taking water into the mouth-swallowing) in the amphibious mudskipper goby, thereby discovering thirst in ray-finned fish. However, roles of dipsogenic/antidipsogenic hormones, which act on the thirst center in terrestrial tetrapods, have remained unclear in the mudskipper thirst. Here we examined the hormonal effects on the mudskipper drinking behaviours, particularly the antagonistic interaction between angiotensin II (AngII) and atrial natriuretic peptide (ANP) which is important for thirst regulation in mammalian 'forebrain'. Expectedly, intracerebroventricular injection of ANP in mudskippers reduced AngII-increased drinking rate. ANP also suppressed the neural activity at the 'hindbrain' region for the swallowing reflex, and the maintenance of buccopharyngeal water due to the swallowing inhibition may attenuate the motivation to move to water. Thus, the hormonal molecules involved in drinking regulation, as well as the influence of buccopharyngeal water, appear to be conserved in distantly related species to solve osmoregulatory problems, whereas hormonal control of thirst at the forebrain might have been acquired only in tetrapod lineage during evolution.

DOI： 10.1038/s41598-019-52870-7

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DOI： 10.1038/s41598-018-24621-7

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その他リンク： http://www.nature.com/articles/s41598-018-24621-7

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記述言語：英語   出版者・発行元：ELSEVIER SCIENCE INC  

The eyestalk hormone, crustacean female sex hormone (CFSH), regulates the development of female secondary sexual characteristics in the blue crab Callinectes sapidus. After its discovery, several CFSH gene orthologs have been identified in some species of the suborder Pleocyemata as well. Similarly, in species of another suborder (Dendrobranchiata), an ortholog (Maj-CFSH) has been characterized as an eyestalk factor expressed in both females and males of the kuruma prawn, Marsupenaeus japonicus. In this study, another novel CFSH isoform was identified in the same species using cDNA cloning, expression analysis, and recombinant protein production. The isoform has "CFSH-family" structural characteristics but is dominantly expressed in the ovary, and was therefore designated as Maj-CFSH-ov. Its mRNA and protein levels in vitellogenic ovaries are higher than those in non-vitellogenic ovaries. In the vitellogenic ovary, both mRNA and protein expression of Maj-CFSH-ov are localized to oogonia and previtellogenic oocytes that occupy a small portion of vitellogenic ovaries, but not to the major developing oocytes. A vitellogenesis-inhibiting peptide of M. japonicus (Pej-SGP-I) reduced the expression of vitellogenin in incubated ovarian fragments, but not that of Maj-CFSH-ov. These results indicate that M. japonicus possesses two CFSH isoforms that are derived from distinct tissues, the central X-organ/sinus gland complex and peripheral ovaries. The expression profile of Maj-CFSH-ov suggests its involvement in some reproductive process other than vitellogenesis.

DOI： 10.1016/j.cbpa.2017.12.009

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記述言語：英語   出版者・発行元：Academic Press Inc.  

In order to better understand the roles that melanocortin receptors (cMCRs) and melanocortin-2 receptor accessory proteins (cMRAP1 and cMRAP2) play in the HPA axis and hypothalamus, adrenal gland and hypothalamus mRNA from 1 day-old white leghorn chicks (Gallus gallus), were analyzed by real-time PCR. mRNA was also made for kidney, ovary, and liver. Mrap1 mRNA could be detected in adrenal tissue, but not in any of the other tissues, and mrap2 mRNA was also detected in the adrenal gland. Finally, all five melanocortin receptors mRNAs could be detected in the adrenal gland
mc2r and mc5r mRNAs were the most abundant. To evaluate any potential interactions between MRAP1 and the MCRs that may occur in adrenal cells, individual chick mcr cDNA constructs were transiently expressed in CHO cells either in the presence or absence of a chick mrap1 cDNA, and the transfected cells were stimulated with hACTH(1–24) at concentrations ranging from 10−13 M to 10−6 M. As expected, MC2R required co-expression with MRAP1 for functional expression
whereas, co-expression of cMC3R with cMRAP1 had no statistically significant effect on sensitivity to hACTH(1–24). However, co-expression of MC4R and MC5R with MRAP1, increased sensitivity for ACTH(1–24) by approximately 35 fold and 365 fold, respectively. However, co-expressing of cMRAP2 with these melanocortin receptors had no effect on sensitivity to hACTH(1–24). Since the real-time PCR analysis detected mrap2 mRNA and mc4r mRNA in the hypothalamus, the interaction between cMC4R and cMRAP2 with respect to sensitivity to ACTH(1–13)NH2 stimulation was also evaluated. However, no effect, either positive or negative, was observed. Finally, the highest levels of mc5r mRNA were detected in liver cells. This observation raises the possibility that in one-day old chicks, activation of the HPA axis may also involve a physiological response from liver cells.

DOI： 10.1016/j.ygcen.2017.09.002

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記述言語：英語  

Thirst aroused in the forebrain by angiotensin II (AngII) or buccal drying motivates terrestrial vertebrates to search for water, whereas aquatic fish can drink surrounding water only by reflex swallowing generated in the hindbrain. Indeed, AngII induces drinking through the hindbrain even after removal of the whole forebrain in aquatic fish. Here we show that AngII induces thirst also in the amphibious mudskipper goby without direct action on the forebrain, but through buccal drying. Intracerebroventricular injection of AngII motivated mudskippers to move into water and drink as with tetrapods. However, AngII primarily increased immunoreactive c-Fos at the hindbrain swallowing center where AngII receptors were expressed, as in other ray-finned fish, and such direct action on the forebrain was not found. Behavioural analyses showed that loss of buccal water on land by AngII-induced swallowing, by piercing holes in the opercula, or by water-absorptive gel placed in the cavity motivated mudskippers to move to water for refilling. Since sensory detection of water at the bucco-pharyngeal cavity like 'dry mouth' has recently been noted to regulate thirst in mammals, similar mechanisms seem to have evolved in distantly related species in order to solve osmoregulatory problems during terrestrialization.

DOI： 10.1038/s41598-017-18611-4

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記述言語：英語   出版者・発行元：Nature Publishing Group  

Nemerteans (ribbon worms) and phoronids (horseshoe worms) are closely related lophotrochozoans - a group of animals including leeches, snails and other invertebrates. Lophotrochozoans represent a superphylum that is crucial to our understanding of bilaterian evolution. However, given the inconsistency of molecular and morphological data for these groups, their origins have been unclear. Here, we present draft genomes of the nemertean Notospermus geniculatus and the phoronid Phoronis australis, together with transcriptomes along the adult bodies. Our genome-based phylogenetic analyses place Nemertea sister to the group containing Phoronida and Brachiopoda. We show that lophotrochozoans share many gene families with deuterostomes, suggesting that these two groups retain a core bilaterian gene repertoire that ecdysozoans (for example, flies and nematodes) and platyzoans (for example, flatworms and rotifers) do not. Comparative transcriptomics demonstrates that lophophores of phoronids and brachiopods are similar not only morphologically, but also at the molecular level. Despite dissimilar head structures, lophophores express vertebrate head and neuronal marker genes. This finding suggests a common origin of bilaterian head patterning, although different heads evolved independently in each lineage. Furthermore, we observe lineage-specific expansions of innate immunity and toxin-related genes. Together, our study reveals a dual nature of lophotrochozoans, where conserved and lineage-specific features shape their evolution.

DOI： 10.1038/s41559-017-0389-y

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記述言語：英語   出版者・発行元：Nature Publishing Group  

Study design: Experimental animal study. Objectives: Although a population of gastrin-releasing peptide (GRP) neurons in the lumbar spinal cord has an important role in erection and ejaculation in rats, little information exists on this GRP system in primates. To identify the male-specific GRP system in the primate spinal cord, we studied the lumbosacral cord in macaque monkeys as a non-human primate model. Setting: University laboratory in Japan. Methods: To determine the gene sequence of GRP precursors, the rhesus macaque monkey genomic sequence data were searched, followed by phylogenetic analysis. Subsequently, immunocytochemical analysis for GRP was performed in the monkey spinal cord. Results: We have used bioinformatics to identify the ortholog gene for GRP precursor in macaque monkeys. Phylogenetic analysis suggested that primate prepro-GRP is separated from that of other mammalian species and clustered to an independent branch as primates. Immunocytochemistry for GRP further demonstrated that male-dominant sexual dimorphism was found in the spinal GRP system in monkeys as in rodents. Conclusion: We have demonstrated in macaque monkeys that the GRP system in the lower spinal cord shows male-specific dimorphism and May have an important role in penile functions not only in rodents but also in primates.

DOI： 10.1038/sc.2017.105

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記述言語：英語   出版者・発行元：Elsevier Ltd  

Corticosteroid receptors are critical for homeostasis maintenance, but understanding of the principal roles of the glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) throughout vertebrates is limited. Lines of constitutive GR-knockout zebrafish and MR-knockout medaka have recently been generated as the first adult-viable corticosteroid receptor-knockout animals, in contrast to the lethality of these receptor knockouts in mice. Here, we describe behavioral and physiological modifications following disruption of corticosteroid receptor function in these animal models. We suggest these data point toward a potentially conserved function of corticosteroid receptors in integrating brain-behavior and visual responses in vertebrates. Finally, we discuss how future work in cartilaginous fishes (Chondrichthyes) will further advance understanding of the unity and diversity of corticosteroid receptor function, since distinct orthologs of GR and MR derived from an ancestral corticoid receptor appear in these basal jawed vertebrates.

DOI： 10.1016/j.jsbmb.2018.02.011

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DOI： 10.1210/en.2018-00043

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記述言語：英語   出版者・発行元：NATURE PUBLISHING GROUP  

To study the critical role of mineralocorticoid signalling, we generated a constitutive mineralocorticoid receptor (MR)-knockout (KO) medaka as the first adult-viable MR-KO animal. This KO medaka displayed abnormal behaviours affected by visual stimuli. In contrast, the loss of MR did not result in overt phenotypic changes in osmoregulation, despite the well-known osmoregulatory functions of MR in mammals. Since glucocorticoid receptor (GR) has been suggested to compensate for loss of MR, we examined expression of duplicated GRs with markedly different ligand sensitivities, in various tissues. qRT-PCR results revealed that the absence of MR induced GR1 in the brain and eyes, but not in osmoregulatory organs. This reinforces the important functions of glucocorticoid signalling, but the minor role of mineralocorticoid signalling, in fish osmoregulation. Because both 11-deoxycorticosterone (DOC) and cortisol are ligands for MR, whereas GRs are specific to cortisol, GR1 signalling may compensate for the absence of cortisol-MR, rather than that of DOC-MR. Thus, this GR expression suggests that our MR-KO model can be used specifically to characterize DOC-MR signalling.

DOI： 10.1038/sdata.2017.189

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：ACTA DERMATO-VENEREOLOGICA  

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記述言語：英語   出版者・発行元：WILEY  

Several regions of the brain and spinal cord control male reproductive function. We previously demonstrated that the gastrin-releasing peptide (GRP) system, located in the lumbosacral spinal cord of rats, controls spinal centers to promote penile reflexes during male copulatory behavior. However, little information exists on the male-specific spinal GRP system in animals other than rats. The objective of this study was to examine the functional generality of the spinal GRP system in mammals using the Asian house musk shrew (Suncus murinus; suncus named as the laboratory strain), a specialized placental mammal model. Mice are also used for a representative model of small laboratory animals. We first isolated complementary DNA encoding GRP in suncus. Phylogenetic analysis revealed that suncus preproGRP was clustered to an independent branch. Reverse transcription-PCR showed that GRP and its receptor mRNAs were both expressed in the lumbar spinal cord of suncus and mice. Immunohistochemistry for GRP demonstrated that the sexually dimorphic GRP system and male-specific expression/distribution patterns of GRP in the lumbosacral spinal cord in suncus are similar to those of mice. In suncus, we further found that most GRP-expressing neurons in males also express androgen receptors, suggesting that this male-dominant system in suncus is also androgen-dependent. Taken together, these results indicate that the sexually dimorphic spinal GRP system exists not only in mice but also in suncus, suggesting that this system is a conserved property in mammals. J. Comp. Neurol. 525:1586-1598, 2017. (c) 2016 Wiley Periodicals, Inc.

DOI： 10.1002/cne.24138

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記述言語：日本語  

J-GLOBAL

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記述言語：英語   出版者・発行元：WILEY  

The crustacean hyperglycemic hormone (CHH) is one of the major hormones in crustaceans, and peptides belonging to the CHH superfamily have been found in diverse ecdysozoans. Although the basic function of CHH is to control energy metabolism, it also plays various roles in crustacean species, such as in molting and vitellogenesis. Here, we present the crystal structure of Pej-SGP-I-Gly, a partially active precursor of CHH from the kuruma prawn Marsupenaeus japonicus, which has an additional Gly residue in place of the C-terminal amide group of the mature Pej-SGP-I. The 1.6-angstrom crystal structure showed not only the common CHH superfamily scaffold comprising three a-helices, three disulfide bridges, and a hydrophobic core but also revealed that the C-terminal part has a variant backbone fold that is specific to Pej-SGP-I-Gly. The a-helix 4 of Pej-SGP-I-Gly was much longer than that of molt-inhibiting hormone (Pej-MIH) from the same species, and as a result, the following C-terminal helix, corresponding to a-helix 5 in MIH, was not formed. Unlike monomeric PejMIH, Pej-SGP-I-Gly forms a homodimer in the crystal structure via its unique a-helix 4. The unexpected dissimilar folds between Pej-SGP-I-Gly and Pej-MIH appear to be the result of their distinct C-terminal amino acid sequences. Variations in amino acid sequences and lengths and the resulting variety of backbone folds allow the C-terminal and sterically adjoining regions to confer different hormonal activities in diverse CHH superfamily members.

DOI： 10.1111/febs.13926

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記述言語：英語   出版者・発行元：NATURE PUBLISHING GROUP  

As in osmoregulation, mineralocorticoid signaling is implicated in the control of brain-behavior actions. Nevertheless, the understanding of this role is limited, partly due to the mortality of mineralocorticoid receptor (MR)-knockout (KO) mice due to impaired Na+ reabsorption. In teleost fish, a distinct mineralocorticoid system has only been identified recently. Here, we generated a constitutive MR-KO medaka as the first adult-viable MR-KO animal, since MR expression is modest in osmoregulatory organs but high in the brain of adult medaka as for most teleosts. Hyper-and hypo-osmoregulation were normal in MR-KO medaka. When we studied the behavioral phenotypes based on the central MR localization, however, MR-KO medaka failed to track moving dots despite having an increase in acceleration of swimming. These findings reinforce previous results showing a minor role for mineralocorticoid signaling in fish osmoregulation, and provide the first convincing evidence that MR is required for normal locomotor activity in response to visual motion stimuli, but not for the recognition of these stimuli per se. We suggest that MR potentially integrates brain-behavioral and visual responses, which might be a conserved function of mineralocorticoid signaling through vertebrates. Importantly, this fish model allows for the possible identification of novel aspects of mineralocorticoid signaling.

DOI： 10.1038/srep37991

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記述言語：英語   出版者・発行元：ELSEVIER IRELAND LTD  

scl is a spontaneous medaka mutant deficient in P450c17I, which is required for production of sex steroids, but not of cortisol, the major role of which is osmoregulation in teleost fish. The scl mutant provides a new model to study the functions of these hormones. We first found that fish homozygous for this mutation have plasma cortisol constitutively at a high physiological level (1000 nM). Since we previously showed that this level reversed the seawater-type differentiation of the medaka gastrointestinal tract, hypoosmoregulation of the scl mutant was analyzed. Muscle water contents in freshwater were normal in scl homozygotes, but the contents were lower than those of the wild type (WT) after seawater transfer. There were no differences in gill mRNA levels of corticosteroid receptors or ion transporters between scl homozygotes and WT. In the intestine, expression of glucocorticoid receptors and Na+/K+/2Cl(-) cotransporter were induced in WT during seawater acclimation, but not in scl homozygotes. The high plasma cortisol may prevent hypoosmoregulation by inhibition of increased intestinal water absorption, essentially by the Na+/K+/2Cl(-) cotransporter, in seawater. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.mce.2016.04.009

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記述言語：英語   出版者・発行元：Elsevier Inc.  

This article contains structure and pharmacological characteristics of melanocortin receptors (MCRs) related to research published in "Characterization of melanocortin receptors from stingray Dasyatis akajei, a cartilaginous fish" (Takahashi et al., 2016) [1]. The amino acid sequences of the stingray, D. akajei, MC1R, MC2R, MC3R, MC4R, and MC5R were aligned with the corresponding melanocortin receptor sequences from the elephant shark, Callorhinchus milii, the dogfish, Squalus acanthias, the goldfish, Carassius auratus, and the mouse, Mus musculus. These alignments provide the basis for phylogenetic analysis of these gnathostome melanocortin receptor sequences. In addition, the Japanese stingray melanocortin receptors were separately expressed in Chinese Hamster Ovary cells, and stimulated with stingray ACTH, α-MSH, β-MSH, γ-MSH, δ-MSH, and β-endorphin. The dose response curves reveal the order of ligand selectivity for each stingray MCR.

DOI： 10.1016/j.dib.2016.04.050

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記述言語：英語   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

Melanocortin (MC) systems are composed of MC peptides such as adrenocorticotropic hormone (ACTH), several molecular forms of melanocyte-stimulating hormones (MSHs) and MC receptors (MCRs). Here we demonstrated that the cartilaginous fish, Dasyatis akajei (stingray) expresses five subtypes of MCR genes-mc1r to mc5r-as in the case of teleost and tetrapod species. This is the first evidence showing the presence of the full repertoire of melanocortin receptors in a single of cartilaginous fish. Expression of respective stingray mcr cDNAs in Chinese hamster ovary cells revealed that Des-acetyl-alpha-MSH exhibited cAMP-producing activity indistinguishable to ACTH(1-24) on MC1R and MC2R, while the activity of Des-acetyl-alpha-MSH on MC3R, MC4R, and MC5R were similar to or slightly greater than that of ACTH(1-24). Notably, in contrast to the other vertebrates, MC2R did not require coexpression with a melanocortin receptor-2 accessory protein 1 (mrap1) cDNA for functional expression. One of the roles of MC system resides in regulation of the pituitary-interrenal (PI) axis-a homologue of tetrapod pituitary adrenal axis. In stingray, interrenal tissues were shown to express mc2r and mc5r as major MCR genes. These results established the presence of functional PI axis in stingray at the level of receptor molecule. While MC2R participates in adrenal functions together with MRAP1 in tetrapod species, the fact that sensitivity of MC5R to Des-acetyl-alpha-MSH and ACTH(1-24) were two order of magnitude higher than MC2R without coexpression with MRAP1 suggested that MC5R could play a more important role than MC2R to transmit signals conveyed by ACTH and MSHs if MRAP1 is really absent in the stingray. (C) 2016 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.ygcen.2016.03.030

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記述言語：英語   出版者・発行元：JAPAN ACAD  

A sexually dimorphic spinal gastrin-releasing peptide (GRP) system in the lumbosacral spinal cord, which projects to the lower spinal centers, controls erection and ejaculation in rats. However, little is known about the postnatal development of this system. In this study, we therefore examined the postnatal development of the male-dominant spinal GRP system and its sexual differentiation in rats using immunohistochemistry. Our results show that male-dominant expression of GRP is prominent from the onset of puberty and that sexually dimorphism persists into adulthood. These results suggest that androgen surge during male puberty plays an important role in the development and maintenance of the male-specific GRP function in the rat spinal cord.

DOI： 10.2183/pjab.92.69

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記述言語：英語   出版者・発行元：BIOMED CENTRAL LTD  

Background: In rats, a sexually dimorphic spinal gastrin-releasing peptide (GRP) system in the lumbosacral spinal cord projects to spinal centers that control erection and ejaculation. This system controls the sexual function of adult males in an androgen-dependent manner. In the present study, we assessed the influence of androgen exposure on the spinal GRP system during a critical period of the development of sexual dimorphism.
Methods: Immunohistochemistry was used to determine if the development of the spinal GRP system is regulated by the perinatal androgen surge. We first analyzed the responses of neonates administered with anti-androgen flutamide. To remove endogenous androgens, rats were castrated at birth. Further, neonatal females were administered androgens during a critical period to evaluate the development of the male-specific spinal GRP system.
Results: Treatment of neonates with flutamide on postnatal days 0 and 1 attenuated the spinal GRP system during adulthood. Castrating male rats at birth resulted in a decrease in the number of GRP neurons and the intensity of neuronal GRP in the spinal cord during adulthood despite testosterone supplementation during puberty. This effect was prevented if the rats were treated with testosterone propionate immediately after castration. Moreover, treating female rats with androgens on the day of birth and the next day, masculinized the spinal GRP system during adulthood, which resembled the masculinized phenotype of adult males and induced a hypermasculine appearance.
Conclusions: The perinatal androgen surge plays a key role in masculinization of the spinal GRP system that controls male sexual behavior. Further, the present study provides potentially new approaches to treat sexual disorders of males.

DOI： 10.1186/s13293-016-0058-x

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記述言語：英語   出版者・発行元：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

Gastrin-releasing peptide (GRP) has recently been identified as an itch-signaling molecule in the primary afferents and spinal cord of rodents. However, little information exists on the expression and localization of GRP in the trigeminal somatosensory system other than in rats. We examined the generality of the trigeminal GRP system in mammals using two distinct species, suncus as a model of specialized placental mammals known to have a well-developed trigeminal sensory system and mice as a representative small laboratory animal. We first analyzed the gross morphology of the trigeminal somatosensory system in suncus to provide a brainstem atlas on which to map GRP distribution. Immunohistochemical analyses showed that 8% of trigeminal ganglion neurons in suncus and 6% in mice expressed GRP. Expression was restricted to cells with smaller somata. The GRP-containing fibers were densely distributed in the superficial layers of the caudal part of the trigeminal spinal nucleus (Vc) but rare in the rostral parts, both in suncus and mice. Expression of GRP receptor mRNA and protein was also detected in the Vc of suncus. Taken together, these results suggest that the trigeminal GRP system mediating itch sensation is conserved in mammals.

DOI： 10.1267/ahc.16030

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記述言語：英語   出版者・発行元：Elsevier Inc.  

This data article contains complementary figure and movies (Supplementary Movies 1-3) related to the research article entitled, "Effective synaptome analysis of itch-mediating neurons in the spinal cord: a novel immunohistochemical methodology using high-voltage electron microscopy" [7]. It is important to show the synaptic connections at the ultrastructural level to understand the neural circuit, which requires the three-dimensional (3-D) analyses in the electron microscopy. Here, we applied a new sample preparation method, a high-contrast en bloc staining according to the protocol of the National Center for Microscopy and Imaging Research (NCMIR), University of California, San Diego, CA, USA to high-voltage electron microscopy (HVEM) tomography in order to examine the 3-D chemical neuroanatomy of the rat spinal cord. Pre-embedding immunoelectron microscopy was used in this study. HVEM has an excellent potential to directly visualize the ultrastructures in semi-thin sections (~5. μm thick), and we have successfully visualized many itch-mediating synaptic connections and neural networks in the spinal cord using "HVEM tomography". Moreover, the methodology used in this study is simple and can be applied in multiple ways. This is an important contribution to ultrastructural investigations of the central nervous system in the present post-genomic age.

DOI： 10.1016/j.dib.2015.07.005

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記述言語：日本語   出版者・発行元：JAPANESE SOC FISHERIES SCIENCE  

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記述言語：英語   出版者・発行元：NATURE PUBLISHING GROUP  

Acclimation from marine to dilute environments constitutes among the dramatic evolutionary transitions in the history of life. Such adaptations have evolved in multiple lineages, but studies of the blood/hemolymph homeostasis mechanisms are limited to those using evolutionarily advanced Deuterostome (chordates) and Ecdysozoa (crustaceans). Here, we examined hemolymph homeostasis in the advanced Lophotrochozoa/mollusc, the other unexplored taxa, and its possible regulation by the vasopressin/oxytocin superfamily peptides known to be implicated in fluid homeostasis in Chordata and Arthropoda. The hemolymph osmotic and ionic status in the euryhaline cephalopod (Octopus ocellatus) following transfer from 30-ppt normal seawater to 20 ppt salinity indicate hyperosmo- and hyperionoregulatory abilities for more than 1 week, as in crustaceans and teleost fish. While ventilation frequency decreased by 1 day, Na+/K+-ATPase activity, which has been generally implicated in ion transport, was induced in two of the eight posterior gills after 1 week. In addition, the octopuses were intravenously injected with 1 or 100 ng/g octopressin or cephalotocin, which are Octopus vasopressin/oxytocin orthologs. After 1 day, octopressin, but not cephalotocin, decreased the hemolymph osmolality and Ca concentrations, as well as urinary Na concentrations. These data provide evidence for possible parallel evolution in hyperionoregulatory mechanisms and coordination by conserved peptides.

DOI： 10.1038/srep14469

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記述言語：英語  

The lack of intravital imaging of axonal transport of mitochondria in the mammalian CNS precludes characterization of the dynamics of axonal transport of mitochondria in the diseased and aged mammalian CNS. Glaucoma, the most common neurodegenerative eye disease, is characterized by axon degeneration and the death of retinal ganglion cells (RGCs) and by an age-related increase in incidence. RGC death is hypothesized to result from disturbances in axonal transport and in mitochondrial function. Here we report minimally invasive intravital multiphoton imaging of anesthetized mouse RGCs through the sclera that provides sequential time-lapse images of mitochondria transported in a single axon with submicrometer resolution. Unlike findings from explants, we show that the axonal transport of mitochondria is highly dynamic in the mammalian CNS in vivo under physiological conditions. Furthermore, in the early stage of glaucoma modeled in adult (4-mo-old) mice, the number of transported mitochondria decreases before RGC death, although transport does not shorten. However, with increasing age up to 23-25 mo, mitochondrial transport (duration, distance, and duty cycle) shortens. In axons, mitochondria-free regions increase and lengths of transported mitochondria decrease with aging, although totally organized transport patterns are preserved in old (23- to 25-mo-old) mice. Moreover, axonal transport of mitochondria is more vulnerable to glaucomatous insults in old mice than in adult mice. These mitochondrial changes with aging may underlie the age-related increase in glaucoma incidence. Our method is useful for characterizing the dynamics of axonal transport of mitochondria and may be applied to other submicrometer structures in the diseased and aged mammalian CNS in vivo.

DOI： 10.1073/pnas.1509879112

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記述言語：英語   出版者・発行元：WILEY-BLACKWELL  

Arginine vasopressin (AVP) is a neurohypophysial hormone synthesized as a part of a prepropeptide precursor containing the signal peptide, AVP hormone, AVP-associated neurophysin II and copeptin in the hypothalamic neurosecretory neurons. A transgenic (Tg) rat line expressing the AVP-eGFP fusion gene has been generated. To establish the AVP-eGFP Tg rat as a unique model for an analysis of AVP dynamics invivo, we first examined the invivo molecular dynamics of the AVP-eGFP fusion gene, and then the release of GFP in response to physiological stimuli. Double immunoelectron microscopy demonstrated that GFP was specifically localized in neurosecretory vesicles of AVP neurons in this Tg rat. After stimulation of the posterior pituitary with high potassium we demonstrated the exocytosis of AVP neurosecretory vesicles containing GFP at the ultrastructural level. Biochemical analyses indicated that the AVP-eGFP fusion gene is subjected to invivo post-translational modifications like the native AVP gene, and is packaged into neurosecretory vesicles as a fusion protein: copeptin(1-14)-GFP. Moreover, GFP release into the circulating blood appeared to be augmented after osmotic stimulation, like native AVP. Thus, here we show for the first time the invivo molecular processing of the AVP-eGFP fusion gene and stimulated secretion after osmotic stimulation in rats. Because GFP behaved like native AVP in the hypothalamo-pituitary axis, and in particular was released into the circulation in response to a physiological stimulus, the AVP-eGFP Tg rat model appears to be a powerful tool for analyzing neuroendocrine systems at the organismal level.

DOI： 10.1111/febs.13291

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記述言語：英語   出版者・発行元：ELSEVIER IRELAND LTD  

Transmission electron microscopy (TEM) is used for three-dimensional (3-D) analysis of synaptic connections in neuroscience research. However, 3-D reconstruction of the synapses using serial ultrathin sections is a powerful but tedious approach requiring advanced technical skills. High-voltage electron microscopy (HVEM) allows examination of thicker sections of biological specimens due to the increased penetration of the more accelerated electrons, which is useful to analyze the 3-D structure of biological specimens. However, it is still difficult to visualize the neural networks and synaptic connections in 3-D using HVEM because of insufficient and non uniform heavy metal staining in the membranous structures in semi-thin sections. Here, we present the successful chemical 3-D neuroanatomy of the rat spinal dorsal horn at the ultrastructural level as a first step for effective synaptome analysis by applying a high-contrast en bloc staining method to immune-HVEM tomography. Our new approach made it possible to examine many itch-mediating synaptic connections and neural networks in the spinal cord simultaneously using HVEM tomography. This novel 3-D electron microscopy is very useful for the analysis of synaptic structure and the chemical neuroanatomy at the 3-D ultrastructural level. (C) 2015 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.neulet.2015.05.031

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記述言語：英語   出版者・発行元：PUBLIC LIBRARY SCIENCE  

The neurohypophysial hormones, arginine vasotocin and isotocin, regulate both hydromineral balance and social behaviors in fish. In the amphibious mudskipper, Periophthalmus modestus, we previously found arginine-vasotocin-specific regulation of aggressive behavior, including migration of the submissive subordinate into water. This migration also implies the need for adaptation to dehydration. Here, we examined the effects of arginine vasotocin and isotocin administration on the amphibious behavior of individual mudskippers in vivo. The mudskippers remained in the water for an increased period of time after 1-8 h of intracerebroventricular (ICV) injection with 500 pg/g arginine vasotocin or isotocin. The 'frequency of migration' was decreased after ICV injection of arginine vasotocin or isotocin, reflecting a tendency to remain in the water. ICV injections of isotocin receptor antagonist with arginine vasotocin or isotocin inhibited all of these hormonal effects. In animals kept out of water, mRNA expression of brain arginine vasotocin and isotocin precursors increased 3- and 1.5-fold, respectively. Given the relatively wide distribution of arginine vasotocin fibres throughout the mudskipper brain, induction of arginine vasotocin and isotocin under terrestrial conditions may be involved also in the preference for an aquatic habitat as ligands for brain isotocin receptors.

DOI： 10.1371/journal.pone.0134605

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記述言語：日本語   出版者・発行元：日本理科教育学会  

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DOI： 10.11457/swsj.69.244

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記述言語：日本語   出版者・発行元：応用生態工学会  

北米産外来種のアメリカザリガニは，在来生物の捕食などを通じて，在来生態系に深刻な影響を及ぼしている。 また，本種が掘る巣穴が水田漏水の原因になるなど水田管理上の問題も引き起こしている。このため，本種は緊急対策外来種と日本の侵略的外来種ワースト 100 に指定され，効率的な駆除手法の開発が求められている。 そこで本研究では，水田水域や河川・湖沼等に定着したアメリカザリガニの駆除に用いる人工巣穴の適したサイズを明らかにすることを目的として，本種による人工巣穴サイズ選好性実験を実施した。灰色の直管型の塩ビ管を人工巣穴とし，内径と長さの異なる人工巣穴をアメリカザリガニに室内水槽内で選択させた。その結果，人工巣穴の内径については，全長（X, mm）と好んで選択された内径（Y, mm）の間に Y＝0.58X＋4.26 という有意な回帰式が得られた。人工巣穴の長さについては，全長の 4 倍の長さの巣穴が好んで選択された。以上の結果をもとに，アメリカザリガニの駆除に用いる体サイズ別の人工巣穴サイズを提案した。

DOI： 10.3825/ece.18.79

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記述言語：日本語  

J-GLOBAL

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記述言語：英語   出版者・発行元：WILEY-BLACKWELL  

Gastrin-releasing peptide (GRP) has recently been identified as an itch-specific neuropeptide in the spinal sensory system in mice, but there are no reports of the expression and distribution of GRP in the trigeminal sensory system in mammals. We characterized and compared GRP-immunoreactive (ir) neurons in the trigeminal ganglion (TG) with those in the rat spinal dorsal root ganglion (DRG). GRP immunoreactivity was expressed in 12% of TG and 6% of DRG neurons and was restricted to the small- and medium-sized type cells. In both the TG and DRG, many GRP-ir neurons also expressed substance P and calcitonin gene-related peptide, but not isolectin B-4. The different proportions of GRP and transient receptor potential vanilloid 1 double-positive neurons in the TG and DRG imply that itch sensations via the TG and DRG pathways are transmitted through distinct mechanisms. The distribution of the axon terminals of GRP-ir primary afferents and their synaptic connectivity with the rat trigeminal sensory nuclei and spinal dorsal horn were investigated by using light and electron microscopic histochemistry. Although GRP-ir fibers were rarely observed in the trigeminal sensory nucleus principalis, oralis, and interpolaris, they were predominant in the superficial layers of the trigeminal sensory nucleus caudalis (Vc), similar to the spinal dorsal horn. Ultrastructural analysis revealed that GRP-ir terminals contained clear microvesicles and large dense-cored vesicles, and formed asymmetric synaptic contacts with a few dendrites in the Vc and spinal dorsal horn. These results suggest that GRP-dependent orofacial and spinal pruriceptive inputs are processed mainly in the superficial laminae of the Vc and spinal dorsal horn. J. Comp. Neurol. 522:1858-1873, 2014. (c) 2013 Wiley Periodicals, Inc.

DOI： 10.1002/cne.23506

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記述言語：英語   出版者・発行元：ZOOLOGICAL SOC JAPAN  

To evaluate the effects of inorganic mercury (InHg) and methylmercury (MeHg) on bone metabolism in a marine teleost, the activity of tartrate-resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP) as indicators of such activity in osteoclasts and osteoblasts, respectively, were examined in scales of nibbler fish (Girella punctata). We found several lines of scales with nearly the same TRAP and ALP activity levels. Using these scales, we evaluated the influence of InHg and MeHg. TRAP activity in the scales treated with InHg (10(-5) and 10(-4) M) and MeHg (10(-6) to 10(-4) M) during 6 hrs of incubation decreased significantly. In contrast, ALP activity decreased after exposure to InHg (10(-5) and 10(-4) M) and MeHg (10(-)6 to 10(-4) M) for 18 and 36 hrs, although its activity did not change after 6 hrs of incubation. As in enzyme activity 6 hrs after incubation, mRNA expression of TRAP (osteoclastic marker) decreased significantly with InHg and MeHg treatment, while that of collagen (osteoblastic marker) did not change significantly. At 6 hrs after incubation, the mRNA expression of metallothionein, which is a metal-binding protein in osteoblasts, was significantly increased following treatment with InHg or MeHg, suggesting that it may be involved in the protection of osteoblasts against mercury exposure up to 6 hrs after incubation. To our knowledge, this is the first report of the effects of mercury on osteoclasts and osteoblasts using marine teleost scale as a model system of bone.

DOI： 10.2108/zs130265

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記述言語：日本語  

J-GLOBAL

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記述言語：英語   出版者・発行元：ELSEVIER IRELAND LTD  

Androgens including testosterone, organize the nervous system as well as masculine external and internal genitalia during the perinatal period. Androgen organization involves promotion of masculine body features, usually by acting through androgen receptors (ARs). We have recently demonstrated that the gastrin-releasing peptide (GRP) system in the lumbar spinal cord also mediates spinal centers promoting penile reflexes during male sexual behavior in rats. Testosterone may induce sexual differentiation of this spinal GRP system during development and maintain its activation in adulthood. In the present study, we examined the role of ARs in the nervous system regulating the development of the sexually dimorphic GRP system. For this purpose, we used a conditional mouse line selectively lacking the AR gene in the nervous system. AR foxed males carrying (mutants) or not (controls) the nestin-Cre transgene were castrated in adulthood and supplemented with physiological amounts of testosterone. Loss of AR expression in the nervous system resulted in a significant decrease in the number of GRP neurons compared to control littermates. Consequently, the intensity of GRP axonal projections onto the lower lumbar and upper sacral spinal cord was greater in control males than in mutant males. These results suggest that ARs expressed in the nervous system play a significant role in the development of the GRP system in the male lumbar spinal cord. The AR-deletion mutation may attenuate sexual behavior and activity of mutant males via spinal GRP system-mediated neural mechanisms. (C) 2013 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.neulet.2013.10.068

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記述言語：日本語   出版者・発行元：一般社団法人 日本科学教育学会  

我々は岡山県において初等・中等教育課程への海洋教育の普及活動を行っている。このような取り組みを行う際に，児童・生徒の海洋に対する関心の実態を把握することは重要となる。そこで今回は，海洋教育の一環として岡山県内外の様々な組織と協力して地域連携による文理横断学習「うなぎ探検隊」を開催し，参加した児童ならびに保護者を対象とした質問紙調査によって海洋に対する関心の実態把握を試みた。調査の結果，学習に対する肯定的意見は多く，海を含めた水辺の環境を保全することの重要性を伝えるという学習の目的は達成された。そして，海洋に対する意識や関心は児童，保護者ともに高いことが明らかになった。一方，参加者の半数が身近な海や川に接する機会がないことも明らかとなった。

DOI： 10.14935/jsser.29.3_79

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記述言語：英語   掲載種別：書評論文，書評，文献紹介等   出版者・発行元：FRONTIERS MEDIA SA  

Prolactin-releasing peptide (PrRP) is one of the RF-amide peptides and was originally identified in the bovine hypothalamus as a stimulator of prolactin (PRL) release. Independently, another RF-amide peptide was found in Japanese crucian carp and named Carassius-RFa (C-RFa), which shows high homology to PrRP and stimulates PRL secretion in teleost fish. Therefore, C-RFa has been recognized as fish PrRP. However, recent work has revealed that PrRP and C-RFa in non-mammalian vertebrates are encoded by separate genes originated through duplication of an ancestral gene. Indeed, both PrRP and C-RFa are suggested to exist in teleost, amphibian, reptile, and avian species. Therefore, we propose that non-mammalian PrRP (C-RFa) be renamed PrRP2. Despite a common evolutionary origin, PrRP2 appears to be a physiological regulator of PRL, whereas this is not a consistent role for PrRP itself. Further work revealed that the biological functions of PrRP and PrRP2 are not limited solely to PRL release, because they are also neuro-modulators of several hypothalamus pituitary axes and are involved in some brain circuits related to the regulation of food intake, stress, and cardiovascular functions. However, these actions appear to be different among vertebrates. For example, central injection of PrRP inhibits feeding behavior in rodents and teleosts, while it stimulates it in chicks. Therefore, both PrRP and PrRP2 have acquired diverse actions through evolution. In this review, we integrate the burgeoning information of structures, expression profiles, and multiple biological actions of PrRP in higher vertebrates, as well as those of PrRP2 in non-mammals.

DOI： 10.3389/fendo.2014.00170

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記述言語：英語   出版者・発行元：Academic Press Inc.  

The hypothalamic hormones, arginine-vasotocin (VT) and isotocin (IT), play central roles in osmoregulation and in the regulation of social behaviors including aggressive behavior in many vertebrates including fish. Here, we examined whether these hormones are associated with aggressive behavior in the mudskipper (Periophthalmus modestus). The mudskipper is an amphibious fish, which lives in the brackish water of river mouths and displays unique aggressive behavior. Upon introduction to each other in an experimental tank with aquatic and terrestrial areas, a pair of males can be classified as aggressive dominant or submissive subordinate based on the frequency of their aggressive acts, which is significantly higher in dominant male. Additionally, the length of stay in terrestrial area of dominant was longer than that of the subordinate. The latter remained in aquatic area almost throughout the period of behavioral observation. The expression of brain VT mRNA was significantly higher in subordinate than in dominant, whereas neither IT mRNA expression nor plasma cortisol level differed between subordinate and dominant male. On the other hand, an intracerebroventricular injection of VT increased aggressive behaviors in mudskippers. In addition to known roles of VT in mediation of aggressive behavior, these results may shed light on the role of endogenous VT toward water migration in submissive mudskippers. The amphibious fish is a valuable experimental model to observe the relationship between effects of central VT on the osmoregulation and social behavioral regulation in vertebrates. © 2013.

DOI： 10.1016/j.ygcen.2013.09.023

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記述言語：英語   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

We have recently identified prolactin (PRL)-releasing peptides (PrRPs) and their stimulating effects on feeding behavior in chicks. To investigate further metabolic functions of PrRP, the present study was performed to clarify whether intracerebroventricular (ICV) injection of PrRP31, an active form of PrRP in chicks, affects heat production (HP), respiratory quotient (RQ) and plasma concentrations of metabolic fuels in chicks. The ICV injection of PrRP31 (94 and 375 pmol) did not affect HP but significantly lowered RQ. The change in RQ implies that PrRP31 shifted the utility of metabolic fuels in the body. This idea was confirmed by subsequent results in which ICV injection of PrRP31 significantly reduced glucose but increased non-esterified fatty acid concentrations in plasma. These shifts in blood metabolic fuels would not be through the increased plasma insulin, because the ICV injection of PrRP31 significantly decreased plasma insulin concentration. On the other hand, ICV injection of another orexigenic peptide, neuropeptide Y (NPY) also induced the insulin release and the metabolic effects were similar to those of PrRP31. Because ICV injection of PrRP31 increased NPY mRNA in the diencephalon, the NPY may mediate the metabolic functions of PrRP31. In summary, the present study suggests that central PrRP31 shifts the utilities of peripheral energy sources, which is not via hyperinsulinemia but via the diencephalon. (C) 2013 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.physbeh.2013.06.017

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記述言語：英語   出版者・発行元：ZOOLOGICAL SOC JAPAN  

Fish scales are a form of calcified tissue similar to that found in human bone. In medaka scales, we detected both osteoblasts and osteoclasts and subsequently developed a new scale assay system. Using this system, we analyzed the osteoblastic and osteoclastic responses under 2-, 3-, and 4-gravity (G) loading by both centrifugation and vibration. After loading for 10 min, the scales from centrifugal and vibration loading were incubated for 6 and 24 hrs, respectively, after which the osteoblastic and osteoclastic activities were measured. Osteoblastic activity significantly increased under 2- to 4-G loading by both centrifugation and vibration. In contrast, we found that osteoclastic activity significantly decreased under 2- and 3-G loading in response to both centrifugation and vibration. Under 4-G loading, osteoclastic activity also decreased on centrifugation, but significantly increased under 4-G loading by vibration, concomitant with markedly increased osteoblastic activity. Expression of the receptor activator of the NF-kappa B ligand (RANKL), an activation factor of osteoclasts expressed in osteoblasts, increased significantly under 4-G loading by vibration but was unchanged by centrifugal loading. A protein sequence similar to osteoprotegerin (OPG), which is known as an osteoclastogenesis inhibitory factor, was found in medaka using our sequence analysis. The ratio of RANKL/OPG-like mRNAs in the vibration-loaded scales was significantly higher than that in the control scales, although there was no difference between centrifugal loaded scales and the control scales. Accordingly, medaka scales provide a useful model by which to analyze bone metabolism in response to physical strain.

DOI： 10.2108/zsj.30.217

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記述言語：英語   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

It has long been held that cortisol, a glucocorticoid in many vertebrates, performs glucocorticoid and mineralocorticoid actions in the teleost fish since it lacks aldosterone. However, in addition to the counterparts of tetrapod mineralocorticoid receptors (MRs), 11-deoxycorticosterone (DOC) has been recently identified as a specific endogenous ligand for the MRs in teleosts. Here, we point out the minor role of mineralocorticoid signaling (i.e., DOC-MR) in the osmoregulation compared with those of glucocorticoid signaling (i.e., cortisol-glucocorticoid receptor [GR]), and review the current findings on the physiological roles of the DOC-MR in teleosts. Cortisol promotes both freshwater and seawater adaptation via the GRs in the osmoregulatory organs such as gills and gastrointestinal tracts, but the expressions of MR mRNA are abundant in the brains especially in the key components of the stress axis and cerebellums. Together with the behavioral effects of intracerebroventricular injection with DOC, the MR is suggested to play an important role in the brain dependent behaviors. Since the abundant expression of central MRs has been reported also in higher vertebrates and the MR is thought to be ancestral to the GR, the role of MR in fish might reflect the principal and original function of corticosteroid signaling. Functional evolution of corticosteroid systems is summarized and areas in need of research like our on-going experiments with MR-knockout medaka are outlined. (C) 2012 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.ygcen.2012.11.016

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記述言語：英語   出版者・発行元：ZOOLOGICAL SOC JAPAN  

DOI： 10.2108/zsj.29.444

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記述言語：英語   出版者・発行元：SPRINGER  

Three-dimensional (3-D) analysis of anatomical ultrastructures is important in biological research. However, 3-D image analysis on exact serial sets of ultra-thin sections from biological specimens is very difficult to achieve, and limited information can be obtained by 3-D reconstruction from these sections due to the small area that can be reconstructed. On the other hand, the high-penetration power of electrons by an ultra-high accelerating voltage enables thick sections of biological specimens to be examined. High-voltage electron microscopy (HVEM) is particularly useful for 3-D analysis of the central nervous system because considerably thick sections can be observed at the ultrastructure level. Here, we applied HVEM tomography assisted by light microscopy to a study of the 3-D chemical neuroanatomy of the rat lower spinal cord annotated by double-labeling immunohistochemistry. This powerful methodology is useful for studying molecular and/or chemical neuroanatomy at the 3-D ultrastructural level.

DOI： 10.1007/s00418-012-0976-6

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記述言語：英語   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

We examined growth of euryhaline Japanese medaka (Oryzias latipes) after transfer to freshwater or seawater from isotonic saline. Growth was unaffected by the different salinities for 1 week, but the body weight increase and BMI of fish kept in freshwater for 2-3 weeks were significantly higher than those in the isotonic controls. These results may reflect the usual habitat of this species. To assess the basis for the difference in growth, energetics and the hepatic stress axis were evaluated 1 week after the transfer. Unexpectedly, despite the higher growth rate, the rate of routine oxygen consumption was significantly higher in freshwater. Plasma cortisol levels in freshwater were significantly higher than those in seawater, and the mRNA levels of the glucocorticoid receptor (GR1) in the liver were significantly lower in freshwater and seawater, compared to that in isotonic saline. Branchial Na+/K+-ATPase activities were also reduced significantly in freshwater and seawater, compared to that in isotonic saline. The higher levels of hepatic GR1 expression and branchial Na+/K+-ATPase activity in isotonic salinity than those in freshwater and seawater for 1 week may account for the lower growth rate under the isotonic condition. After 3 weeks, however, the Na+/K+-ATPase activity in seawater was significantly higher than that in freshwater. No significant difference in growth rate between freshwater and seawater groups indicates that medaka is a good model for studies of hypo- and hyperosmotic adaptations, since osmoregulation is not strongly associated with size and growth. (C) 2012 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.ygcen.2012.05.001

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記述言語：英語   出版者・発行元：ZOOLOGICAL SOC JAPAN  

Using our original in vitro assay system with goldfish scales, we examined the direct effect of prostaglandin E-2 (PGE(2)) on osteoclasts and osteoblasts in teleosts. In this assay system, we measured the activity of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) as respective indicators of each activity in osteoblasts and osteoclasts. ALP activity in scales significantly increased following treatment at high concentration of PGE(2) (10(-7) and 10(-6) M) over 6 hrs of incubation. At 18 hrs of incubation, ALP activity also significantly increased in the PGE(2) (10(-9) to 10(-6) M)-treated scale. In the case of osteoclasts, TRAP activity tended to increase at 6 hrs of incubation, and then significantly increased at 18 hrs of incubation by PGE(2) (10(-7) to 10(-6) M) treatment. At 18 hrs of incubation, the mRNA expression of osteoclastic markers (TRAP and cathepsin K) and receptor activator of the NF-kappa B ligand (RANKL), an activating factor of osteoclasts expressed in osteoblasts, increased in PGE(2) treated-scales. Thus, PGE(2) acts on osteoblasts, and then increases the osteoclastic activity in the scales of goldfish as it does in the bone of mammals. In an in vivo experiment, plasma calcium levels and scale TRAP and ALP activities in the PGE(2)-injencted goldfish increased significantly. We conclude that, in teleosts, PGE(2) activates both osteoblasts and osteoclasts and participates in calcium metabolism.

DOI： 10.2108/zsj.29.499

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記述言語：英語   出版者・発行元：ELSEVIER SCIENCE BV  

In cartilaginous fish, two cDNAs encoding calcitonin-family receptors were isolated for the first time from the stingray brain. The open reading frame of one receptor cDNA coded a 525-amino acid protein. The amino acid identity of this receptor to human calcitonin-receptor-like receptor (CRLR) is 64.5%, frog CRLR is 64.7%, and flounder CRLR is 61.2% and this was higher than to human calcitonin receptor (CTR) (46.1%), frog CTR (54.7%), and flounder CTR (48.9%). We strongly suggested that this receptor is a ray CRLR based on phylogenetic analysis. In case of the second receptor, amino acid identity among CRLRs (human 50.5%, frog 50.7%, flounder 48.0%) and CTRs (human 43.2%, frog 49.1%, flounder 41.8%) was similar. From phylogenetic analysis of both CRLRs and CTRs, we believe that this receptor is ray CTR. The expression of ray CRLR mRNA was predominantly detected in the nervous system (brain) and vascular system (atrium, ventricle, and gill), which reflects the similar localization of CGRP in the nervous and vascular systems as mammals. It was observed that the second receptor was expressed in several tissues, namely cartilage, brain, pituitary gland, gill, atrium, ventricle, pancreas, spleen, liver, gall bladder, intestine, rectal gland, kidney, testis and ovary. This localization pattern was very similar to flounder CTR. Both receptor mRNAs were strongly expressed in the gill. This suggests that the calcitonin-family members are involved in the osmoregulation of stingray as this fish is known to be euryhaline. When a stingray was transferred to diluted seawater (20% seawater), the expression of both receptors significantly decreased in the gill. Similar results were obtained in the kidney of the stingray. Thus, our cloning and isolation of both receptors in the stingray will be helpful for elucidation of their physiological role(s) such as osmoregulation including calcium metabolism of cartilaginous fish. (C) 2012 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.gene.2012.03.042

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記述言語：英語   出版者・発行元：FRONTIERS IN BIOSCIENCE INC  

Steroid hormones easily cross the blood-brain barrier because of their physicochemical lipid solubility. The hormones act through nuclear receptor-mediated mechanisms and modulate gene transcription. In contrast to their genomic actions, the non-genomic rapid action of steroid hormones, acting via various types of membrane-associated receptors, reveals pharmacological properties that are distinct from the actions of the intracellular nuclear receptors. As a result, non-genomic rapid actions have gained increased scientific interest. However, insight into the phylogenic and/or comparative actions of steroids in the brain is still poorly understood. In this review, we summarize recent findings concerning the rapid, non-genomic signaling of steroid hormones in the vertebrate central nervous system, and we discuss (using a comparative view from fish to mammals) recently published data regarding the mechanism underlying physiology and behavior.

DOI： 10.2741/3970

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© 2012, Nanba et al.; licensee Springer. Osmo- and ionoregulatory abilities were examined in the intertidal grapsid crab, Gaetice depressus, transferred from normal seawater (30 ppt) to low (10 ppt) or high (50 ppt) salinities for 2 and 10 days, in addition to animals kept out of water for 2 days. The results of the hemolymph osmotic and ionic status indicate that G. depressus is able to adapt for more than 10 days in these salinities and for 2 days under terrestrial conditions. Especially, the free Ca2+ concentration was relatively maintained compared with concentrations of monovalent ions and osmolality values in 10 and 50 ppt, partly using the complexed calcium (total minus free calcium) as an internal reserve in the hemolymph. In 10 ppt, complexed calcium disappeared from the hemolymph after 10 days, indicating that all the hemolymph calcium was ionized. In 50 ppt, free Ca2+ was regulated to lower levels than concentrations in the medium, while total calcium increased to higher levels after 2 days. Examination of Na+/K+-ATPase activity, which has been implicated in ion transport in many crustaceans, revealed that induction of high Na+/K+-ATPase activity varies among the posterior gills in response to salinities. Ten-ppt salinity induces activity in two of the posterior gills (gill numbers 6 and 7, eight in total), albeit with differing degrees of response. In contrast, 50-ppt salinity stimulates the activity primarily in gill number 8, suggesting that this gill may be associated specifically with ion excretion in G. depressus. As a euryhaline amphibious crab, this abundant species around Japan will serve as a model to study the osmotic/ionic regulatory mechanisms which operate in crustaceans.

DOI： 10.1186/2008-6970-4-18

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記述言語：英語   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

It has long been held that cortisol, a glucocorticoid in many vertebrates, carries out both glucocorticoid and mineralocorticoid actions in teleost fish. However, 11-deoxycorticosterone (DOC) has been identified as a specific endogenous ligand for the teleostean mineralocorticoid receptor (MR). Furthermore, the expressions of MR mRNA are modest in the osmoregulatory organs, but considerably higher in the brain of most teleosts. These recent findings suggest that the mineralocorticoid system (DOC/MR) may carry out some behavioral functions in fish. To test this possibility, we examined the effects of cortisol and DOC administration in the amphibious behavior in mudskipper (Periophthalmus modestus) in vivo. It was found that mudskippers remained in the water for an increased period of time when they were immersed into 5 mu M DOC or cortisol for 8 h. Additionally, an exposure to 25 mu M DOC for 4 to 8 h caused a decreased migratory frequency of mudskippers to the water, reflected a tendency to remain in the water. It was further observed that after 8 h of intracerebroventricular (ICV) injection with 0.3 pmol DOC or cortisol the staying period in the water increased in fish. The migratory frequency was decreased after ICV DOC injection which indicated that fishes stayed in the water. Concurrent ICV injections of cortisol with RU486 [a specific glucocorticoid-receptor (GR) antagonist] inhibited only the partial effects of cortisol. Together with no changes in the plasma DOC concentrations under terrestrial conditions, these results indicate the involvement of brain MRs as cortisol receptors in the preference for an aquatic habitat of mudskippers. Although the role of GR signaling cannot be excluded in the aquatic preference, our data further suggest that the MR may play an important role in the brain dependent behaviors of teleost fish. (C) 2011 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.physbeh.2011.06.002

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記述言語：英語   出版者・発行元：SPRINGER TOKYO  

Many teleost fishes in lowland fresh waters spawn in ephemeral flooded areas, the bottoms of which are prone to hypoxia. Little is known about how embryos and larvae deal with these potentially hostile environments. This study examines the functional and behavioral ontogeny of one such species, the kissing loach (Parabotia curta). Kissing loach eggs are demersal and adhesive. Hatching occurs at 24.8 +/- A 0.1 h post-fertilization at 25A degrees C, much earlier than most fish species. The newly hatched larvae are precocious with no functional mouth, fins or eye pigmentation. Swimbladder inflation normally occurs at about 4 days posthatch, even before which the hatched larvae moved immediately toward the water surface to hang from water moss. Experiments with larvae 20 h after hatching showed that they spent significantly less time on the bottom in hypoxic water (2 mg/l) than in normoxic water, and suggest that hypoxia is a major directive factor in eliciting surfacing behavior. For the kissing loach, we have previously reported short-term spawning after the formation of flood areas as well as wide scattering of the spawned eggs in the temporal flooded areas. These traits with the present results of hatching at an early stage and the immediate upward movement of larvae are considered to be effective strategies for using ephemeral, hypoxic flooded areas for reproduction.

DOI： 10.1007/s10228-011-0217-3

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記述言語：英語   出版者・発行元：WILEY-BLACKWELL PUBLISHING, INC  

Prolactin (PRL)-releasing peptides (PrRP) have been identified in mammals, amphibians and fishes, and these animals have several PrRPs that consist of different numbers of amino acids such as 20, 31 and 37. In the present study, we identified the cDNA encoding chicken prepro-PrRP, which can generate putative PrRPs, and cloned and sequenced it. Sequences for the coding region suggested the occurrence of putative PrRPs of 20, 31 and 32 amino acid residues. The amino acid sequence of chicken PrRP20 showed 100%, 95% and 70% identity with those of PrRP20s from teleosts, Xenopus laevis and mammals, respectively. On the other hand, chicken PrRP31 showed approximately 90% and 52-55% homology to PrRP31s of X. laevis and mammals, respectively. Native chicken PrRPs were purified from an acid extract of chick brain by a Sep-Pak C18 cartridge (Waters Corp., Milford, MA, USA), affinity chromatography using anti-salmon PrRP serum, and reverse phase high-performance liquid chromatography (HPLC) on an ODS-120T column (TOSOH, Tokyo, Japan). The existence of chicken PrRP20 and PrRP31 in the brain was demonstrated by comparing them with the synthetic peptides using HPLC and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. Chicken PrRP31 increased plasma PRL concentration when administered peripherally, whereas central administration decreased the concentration, suggesting that chicken PrRP31 has a distinct effect on PRL secretion between tissues in chicks. On the other hand, plasma growth hormone concentration decreased with both peripheral and central administrations of chicken PrRP31. Furthermore, central administration of chicken PrRP31 increased food intake in chicks compared to those observed in mammals and fishes. Taken together with the results indicating that chicken PrRP20 did not show endocrine and behavioural effects, we showed that chicken PrRP has a similar amino acid sequence to teleosts, Xenopus laevis and mammals, although the actions were variable among vertebrates.

DOI： 10.1111/j.1365-2826.2010.02078.x

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記述言語：英語   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

Aims: Cortisol is a glucocorticoid in mammals, but has both gluco- and mineralocorticoid activities in teleost fish. Our previous in vivo studies on osmoregulatory esophagi of euryhaline fish showed that epithelial apoptosis for the simple epithelium in seawater and cell proliferation for the stratified epithelium in fresh water are both induced by cortisol. The aim of the present study was to examine the mechanism of these dual cortisol effects on esophageal cell turnover.
Main methods: We developed a tissue culture method for the esophagus from euryhaline medaka (Oryzias latipes) and assessed cell proliferation and apoptosis in vitro in response to cortisol and 11-deoxycorticosterone (DOC), a recently identified agonist of the teleostean mineralocorticoid receptor.
Key findings: Epithelial apoptosis, a well-established glucocorticoid function, was stimulated by treatment of the esophagus culture with 10 nM cortisol for 8 days, but no effects were seen at higher doses (100 and 1000 nM). In contrast, cell proliferation was induced by 1000 nM cortisol treatment for 8 days and this response was dose-dependent. Both effects were blocked by RU-486, a glucocorticoid receptor antagonist. DOC showed no significant effects at 10-1000 nM.
Significance: In the esophageal epithelium in euryhaline fish, cortisol induces either apoptosis or cell proliferation via the glucocorticoid receptor, depending on the cortisol concentration. The glucocorticoid signaling may play a more important role than mineralocorticoid signaling in differentiation of the osmoregulatory esophagus in euryhaline fishes. (C) 2010 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.lfs.2010.11.017

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記述言語：英語   出版者・発行元：ELSEVIER SCIENCE BV  

Iodine-131 (physical half-life: 8.04 days) was detected in brown algae collected off the Japanese coast. Brown algae have been extensively used as bioindicators for radioiodine because of their ability to accumulate radionuclides in high concentration factors. The maximum measured specific activity of (131)I in brown algae was 0.37 +/- 0.010 Bq/kg-wet. Cesium-137 was also detected in all brown algal samples used in this study. There was no correlation between specific activities of (131)I and (137)Cs in these seaweeds. The specific activity of (137)Cs ranged from 0.0034 +/- 0.00075 to 0.090 +/- 0.014 Bq/kg-wet. Low specific activity and minimal variability of (137)Cs in brown algae indicated that past nuclear weapon tests were the source of (137)Cs. Although nuclear power stations and nuclear fuel reprocessing plants are known to be pollution sources of (131)I, there was no relationship between the sites where (131)I was detected and the locations of nuclear power facilities. Most of the sites where (131)I was detected were near big cities with large populations. Iodine-131 is frequently used in diagnostic and therapeutic nuclear medicine. On the basis of the results, we suggest that the likely pollution source of (131)I, detected in brown seaweeds, is not nuclear power facilities, but nuclear medicine procedures. (C) 2010 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.scitotenv.2010.04.001

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DOI： 10.1016/j.scitotenv.2010.04.001

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記述言語：英語   出版者・発行元：ZOOLOGICAL SOC JAPAN  

In teleosts, prolactin is involved in calcium regulation, but its role in scale/bone metabolism is unclear. Using the in-vitro system with goldfish scales developed recently, we explored the effects of teleost prolactin, growth hormone, and somatolactin on osteoclasts and osteoblasts. Addition of prolactin at concentrations of 0.01-100 ng/ml reduced osteoclastic activity, partly via osteoclast apoptosis, after 6-18 h incubation. Conversely, growth hormone and somatolactin at a concentration of 100 ng/ml increased osteoclastic activity after 18 h incubation, indicating the specificity of the inhibitory effect of prolactin on osteoclastic activity. On the other hand, these three hormones promoted osteoblastic activity at concentrations of 10-100 ng/ml. The results from this study are the first demonstration of direct effects of prolactin on scale/bone metabolism and osteoclastic activity in a teleost.

DOI： 10.2108/zsj.25.739

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記述言語：英語   出版者・発行元：ZOOLOGICAL SOC JAPAN  

The natural spawning behavior of the kissing loach, an endangered species of Botiidae, was investigated in the wild in early June for two years in relation to several environmental factors. Kissing loaches spawned in temporary waters after elevation in water level. All spawnings observed (n=163) occurred within 3-5.5 hours from late afternoon to night after formation of the temporary water. These spawnings were performed by one female and one (71%) or two (29%) males in densely vegetated lentic waters. The female and following male(s) swam into dense grasses, where they vibrated to spawn intermittently. After the vibration continuing for 3-20 seconds, they moved to other parts of the dense grassy area and began vibration again. This sequence of spawning behavior was usually repeated several times, and the eggs were thus scattered widely. The spawning behavior and the rapid larval development of this species appear to be adaptations for the use of temporary waters as a spawning ground. The rise in water level and the consequent formation of temporary waters appear to be crucial triggers for reproduction of the kissing loach.

DOI： 10.2108/zsj.24.850

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記述言語：英語   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

We have previously shown that esophageal epithelium of a euryhaline goby displays elevated cell proliferation in freshwater (FW) fish, but undergo apoptosis during seawater (SW) acclimation. Prolactin (PRL) injection into the goby induced the cell proliferation, whereas cortisol treatment stimulated the cell proliferation and apoptosis [Takahashi, H., Sakamoto, T., Narita, K., 2006a. Cell proliferation and apoptosis in the anterior intestine of an amphibious, euryhaline mudskipper (Periophthalmus modestus). J. Comp. Physiol. B 176, 463-468, 2006). In the euryhaline tilapia (Oreochromis mossambicus), the dynamics of changes in cortisol-glucccorticoid receptors (GR) during acclimation to different salinities also suggests a role for glucocorticoid signaling in the esophageal cell turnover, but the mode of PRL action remains largely unclear. In the present study, we report on effects in the tilapia esophagus that result from changes in environmental salinity. Specifically, we assessed: (1) mRNA expression of PRL receptor (PRLR) using quantitative real-time RTPCR; (2) esophageal cell proliferation and apoptosis, using immunohistochemistry of proliferating cells nuclear antigen (PCNA) and in situ nick end-labeling of genomic DNA (TUNEL); (3) the possible localization of immunoreactive PRLR on proliferating/apoptotic cells. Plasma PRL increased after FW acclimation; PRLR mRNA levels in the esophagus of FW fish were significantly higher than those in SW-acclimated fish. Cell proliferation was induced randomly throughout the esophageal epithelium after FW acclimation, while cell division and increased apoptosis were concentrated at the tips of esopliageal epithelial folds in SW-acclimated fish. Immunoreactive PRLR appeared to be localized at proliferating cells and at certain apoptotic cells, whereas immunoreactive GR was observed over the whole epithelium including the apoptotic/proliferating cells. Thus, PRL appears to affect cell turnover directly in the esophageal epithelium of the eurylialine tilapia. (c) 2007 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.ygcen.2007.02.021

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DOI： 10.2108/zsj.24.922

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記述言語：英語   出版者・発行元：ELSEVIER SCIENCE INC  

Amino acid sequences for identified prolactin (PRL)-releasing peptides (PrRPs) were conserved in mammals (> 90%) or teleost fishes (100%), but there were considerable differences between these classes in the sequence (< 65%) as well as in the role of PrRP. In species other than fishes and mammals, we have identified frog PrRP. The cDNA encoding Xenopus laevis prepro-PrRP, which can generate putative PrRPs, was cloned and sequenced. Sequences for the coding region showed higher identity with teleost PrRPs than mammalian homologues, but suggested the occurrence of putative PrRPs of 20 and 31 residues as in mammals. The amino acid sequence of PrRP20 was only one residue different from teleost PrRP20, but shared 70% identity with mammalian PrRP20s. In primary cultures of bullfrog (Rana catesbeiana) pituitary cells, Xenopus PrRPs increased prolactin concentrations in culture medium to 130-160% of the control, but PrRPs was much less potent than thyrotropin-releasing hormone (TRH) causing a three- to four-fold increase in prolactin concentrations. PrRP mRNA levels in the developing Xenopus brain peak in early prometamorphosis, different from prolactin levels. PrRP may not be a major prolactin-releasing factor (PRF), at least in adult frogs, as in mammals. (c) 2006 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.peptides.2006.08.003

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記述言語：英語   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

We have previously shown that anterior intestinal epithelium of the euryhaline mudskipper (Periophthalmus modestus) undergoes apoptosis during seawater (SW) acclimation, whereas elevated cell proliferation was observed in freshwater (FW)-acclimated fish. To understand the possible endocrine regulation of the gastrointestinal cell turnover during salinity acclimation, we examined the ratios of apoptotic and proliferating cells in the anterior intestine of one-third SW-acclimated mudskipper treated with triiodothyronine (T-3) cortisol, 11-deoxycorticosterone (DOC, the putative teleostean mineralocorticoid), or prolactin (PRL). In situ nick end labeling of genomic DNA (TUNEL) and immunohistochemistry of proliferating cells nuclear antigen (PCNA) were used as indicators of apoptosis and cell proliferations, respectively. Cortisol significantly elevated apoptosis (P < 0.05) in the epithelia and connective tissues and also stimulated the epithelial cell proliferation (P < 0.05). PRL induced epithelial cell proliferation (P < 0.05), but did not affect apoptotic status of the intestinal epithelium. Neither T-3 nor DOC had any impact on cell proliferation or apoptosis. Together, our results suggest a role for cortisol and PRL in the regulation of anterior intestinal epithelial turnover during salinity acclimation in this species. (c) 2006 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.lfs.2006.06.021

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記述言語：英語   出版者・発行元：SPRINGER  

In order to replace the diffusive loss of water to the surrounding environment, seawater (SW)-acclimated euryhaline fishes have gastrointestinal tracts with higher ion/water flux in concert with greater permeability, and contrast that to freshwater (FW)-acclimated fish. To understand the cellular basis for these differences, we examined cell proliferation and apoptosis in the anterior intestine of mudskipper transferred from one-third SW to FW or to SW for 1 and 7 days, and those kept out of water for 1 day. The intestinal apoptosis (indicated by DNA laddering) increased during seawater acclimation. TUNEL staining detected numerous apoptotic cells over the epithelium of SW-acclimated fish. Cell proliferation ([H-3]thymidine incorporation) in the FW fish was greater than those in SW 7 days after transfer. Labeling with a Proliferating cell nuclear antigen (PCNA) antibody indicated that proliferating cells were greater in number and randomly distributed in the epithelium of FW fish, whereas in SW fish they were almost entirely in the troughs of the intestinal folds. There were no changes in cell turnover in fish kept out of water. During acclimation to different salinities, modification of the cell turnover and abundance may play an important role in regulating the permeability (and transport capacity) of the gastrointestinal tract of fish.

DOI： 10.1007/s00360-006-0067-x

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記述言語：英語   出版者・発行元：ELSEVIER SCIENCE INC  

We administered prolactin-releasing peptide (PrRP) or anti-PrRP antiserum to goldfish in, fresh water and analyzed their effects on prolactin and osmoregulatory mechanisms. The pituitary mRNA level of prolactin increased by PrRP but decreased by anti-PrRP. The rate of water inflow in the gills decreased by PrRP and increased by anti-PrRP, showing that PrRP restricts branchial water permeability, as also restricted by prolactin. PrRP also expanded the mucous cell layers on the scales, which may restrict efficiently water inflow by the mucous system. Eventually, the plasma osmotic pressure decreased by anti-PrRP. We conclude that PrRP is essential to maintain prolactin levels and osmotic balance in fresh water. (c) 2006 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.peptides.2005.06.034

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記述言語：英語   出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD  

Cortisol plays an important role in controlling intestinal water and ion transport in teleosts possibly through glucocorticoid receptor (GR) and/or mineralocorticoid receptor. To better understand the role of GR in the teleost intestine, in a euryhaline tilapia, Oreochromis mossambicus, we examined (1) the intestinal localizations of GR; (2) the effects of environmental salinity challenge and cortisol treatment oil GR mRNA expression. The mRNA abundance of GR in the posterior intestinal region of tilapia was found to be higher than that in the anterior and middle intestine. In the posterior intestine, GR appears to be localized in the mucosal layer. GR mRNA levels in the posterior intestine were elevated after exposure of freshwater fish to seawater for 7 days following an increase in plasma cortisol. Similarly, cortisol implantation in freshwater tilapia for 7 days elevated the intestinal GR mRNA. These results indicate that seawater acclimation is accompanied by upregulation of GR mRNA abundance in intestinal tissue, possibly as a consequence of the elevation of cortisol levels. In contrast, a single intraperitoneal injection of cortisol into freshwater tilapia decreased intestinal GR mRNA. This downregulation of the GR mRNA by cortisol suggests a dual mode of autoregulation of GR expression by cortisol. (c) 2005 Elsevier Inc. All lights reserved.

DOI： 10.1016/j.lfs.2005.09.050

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DOI： 10.1016/j.ygcen.2005.10.008

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（全国大会，その他学術会議）  

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記述言語：英語   出版者・発行元：SPRINGER  

Mugilogobius abei has the ability to produce large amounts of urea when exposed to high ambient ammonia. Despite this metabolically costly approach, and reports of growth inhibition effects of ammonia on fish, M. abei exposed to ammonia shows no adverse effects on growth. To investigate this observation the growth of M. abei was measured at room temperatures for 8 weeks at a constant ration level under solitary and grouped conditions, in 20% SW with or without (control) 2 mM NH4Cl. Furthermore, pituitary mRNA levels of growth hormone, oxygen consumption, incorporation of external N-15-ammonia into amino acid and protein fractions as well as behavioral activities were also examined. The specific growth rates of ammonia-exposed fish under grouped condition over the 8 weeks were significantly higher than those of control, while those rates under solitary condition were not significantly different between the treatments. The pituitary of ammonia-exposed fish had higher growth hormone mRNA than in control fish. The use of N-15 isotope revealed that M. abei can actively use external ammonia as a supplementary nitrogen source. Oxygen consumption of ammonia-exposed fish was significantly lower than that of control fish. Locomotor activity and aggressive behavior under grouped condition were significantly reduced in ammonia-exposed fish as compared to those of control. These combined alterations in the ammonia-exposed fish may result in the higher growth rates.

DOI： 10.1007/s00360-005-0001-7

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記述言語：英語   出版者・発行元：SOC ENDOCRINOLOGY  

Prolactin (PRL)-releasing peptide (PrRP) is a strong candidate stimulator of pituitary PRL transcription and secretion in teleosts. However, the role in control of extrapituitary PRL expression is unclear even in mammals. To study the possible presence of PrRP-PRL axes not only in the brain-pituitary but also in peripheral organs, the expression patterns of PrRP, PRL and growth hormone (GH) were characterized in amphibious euryhaline mudskippers (Periophthalmus modestus). PrRP mRNA is abundantly expressed not only in the brain but also in the liver, gut and ovary, while less abundant expression was also detected in the skin and kidney. Corresponding to the distribution of PrRP mRNA, PRL mRNA was also detectable in these organs. During adaptation to different environments, the changes in mRNA levels of PrRP paralleled those in PRL in the brain-pituitary, liver and gut in an organ-specific manner. Brain PrRP mRNA and the pituitary PRL mRNA increased under freshwater and terrestrial conditions (P < 0.05); expression of PrRP and PRL in the gut of freshwater fish was higher (P < 0.05) than those in sea-water fish although there were no changes in fish kept out of water; no significant change was seen in the liver. Expressions of GH were not correlated with PrRP. In the gut, PrRP and PRL appear to be co-localized in the mucosal layer, especially in the mucous cells. Thus, PrRP may also be a local modulator of extrapituitary PRL expression and the PrRP-PRL axes in various organs may play an organ-specific role during environmental adaptation.

DOI： 10.1677/jme.1.01768

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DOI： 10.1196/annals.1327.023

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記述言語：英語   出版者・発行元：SPRINGER-VERLAG  

Prolactin-releasing peptide (PrRP), recently isolated from the brain of mammals and teleosts, is a strong candidate for being a stimulatory hormone of pituitary prolactin secretion. The present study examined whether or not PrRP is capable of inducing prolactin gene expression and elevating plasma prolactin levels in vivo in cannulated rainbow trout. Following a single intra-arterial injection of chum salmon PrRP (40 nmol kg(-1)) through a dorsal aorta catheter, plasma prolactin levels increased (P<0.05) rapidly (2 min and 30 min), and prolactin mRNA levels were elevated (P<0.05) in pituitaries sampled 8 h after the injection. In contrast, plasma levels of somatolactin were decreased (P<0.05) and growth hormone and somatolactin mRNA levels were not significantly affected by PrRP. Thus, PrRP appears to be a potent prolactin secretagogue as well as prolactin transcription inducer in vivo in the rainbow trout.

DOI： 10.1007/s00360-003-0340-1

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DOI： 10.1016/S0044-8486(03)00124-8

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DOI： 10.1016/S0074-7696(05)25003-9

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記述言語：英語   出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE  

In the tilapia (Oreochromis mossambicus), as in many teleosts, prolactin (PRL) plays a major role in osmoregulation in freshwater. Recently, PRL-releasing peptides (PrRPs) have been characterized in mammals. Independently, a novel C-terminal RF (arginine-phenylalanine) amide peptide (Carrasius RF amide: C-RFa), which is structurally related to mammalian PrRPs, has been isolated from the brain of the Japanese crucian carp. The putative PrRP was purified from an acid extract of tilapia brain by affinity chromatography with antibody against synthetic C-RFa and HPLC on a reverse-phase ODS-120 column. The tilapia PrRP cDNA was subsequently cloned by polymerase chain reaction. The cDNA consists of 619 bp encoding a preprohormone of 117 amino acids. Sequence comparison of the isolated peptide and the preprohormone revealed that tilapia PrRP contains 20 amino acids and is identical to C-RFa. Incubation of the tilapia pituitary with synthetic C-RFa (100 nM) significantly stimulated the release of two forms of tilapia PRL (PRI188 and PRI177). However, the effect of C-RFa was less pronounced than the marked increase in PRL release in response to hyposmotic medium. The ability of C-RFa to stimulate PRL release appears to be specific, since C-RFa failed to stimulate growth hormone release from the pituitary in organ culture. In contrast, rat and human PrRPs had no effect on PRL release. C-RFa was equipotent with chicken GnRH in stimulating PRL release in the pituitary preincubated with estradiol 17beta. Circulating levels of PRL were significantly increased 1 h after intraperitoneal injection of 0.1 mug/g of C-RFa in female tilapia in freshwater but not in males. These results suggest that C-RFa is physiologically involved in the control of PRL secretion in tilapia. (C) 2002 Elsevier Science (USA).

DOI： 10.1006/gcen.2001.7727

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DOI： 10.1007/s00360-002-0272-1

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DOI： 10.2108/zsj.20.1497

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DOI： 10.1007/s00360-002-0274-z

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DOI： 10.1016/S0044-8486(00)00471-3

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DOI： 10.2108/zsj.18.1163

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DOI： 10.2108/zsj.18.1163

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DOI： 10.1677/joe.0.1700227

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DOI： 10.1002/(SICI)1097-010X(20000501)286:6<666::AID-JEZ14>3.0.CO;2-G

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DOI： 10.1023/A:1007843508043

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記述言語：英語   出版者・発行元：SOC ENDOCRINOLOGY  

We examined the effects of environmental salinity on circulating levels of the two prolactins (tPRL(177) and tPRL(188)) and levels of pituitary tPRL(177) and tPRL(188) mRNA in the euryhaline tilapia, Oreochromis mossambicus. Fish were sham-operated or hypophysectomized and the rostral pars distalis (RPD) autotransplanted onto the optic nerve. Following post-operative recovery in 1/4 seawater, tilapia were transferred to fresh water (FW), 1/4 seawater (SW) or SW. Serum tPRL(177) and tPRL(188) levels in sham-operated and RPD-autotransplanted fish were highest in FW and decreased as salinity was increased. tPRL(177) and tPRL(188) mRNA levels in RPD implants as well as in pituitaries from the sham-operated fish were also highest in FW and decreased with increasing salinity. Serum osmolality increased with salinity, with the highest levels occurring in die seawater groups. We conclude that some plasma factor (probably plasma osmolality), in the absence of hypothalamic innervation, exerts a direct regulatory action on prolactin release and gene expression in the pituitary of O. mossambicus. This regulation is in accord with the actions of the two prolactins in the freshwater osmoregulation of the tilapia.

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DOI： 10.2108/zsj.16.775

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記述言語：英語  

DOI： 10.2108/zsj.15.205

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記述言語：英語   出版者・発行元：Zoological Society of Japan  

Novel clonidine binding sites were characterized in the intestinal membrane isolated from seawater eels. The specific clonidine binding sites consisted of at least two classes, high affinity (Kd = 1.4 ± 0.3 nM, n = 5) and low affinity (Kd = 175 ± 34 nM, n = 5) sites. The specific binding of 2 nM [3H]clonidine was most enhanced at 20°C and at pH 7.5, and reversed by unlabelled clonidine. Such binding was hardly inhibited by adrenaline, yohimbine or rauwolscine, indicating that most binding sites are distinct from α2-adrenoceptor. The specific clonidine binding was inhibited by various imidazoline/guanidinium drugs, indicating existence of imidazoline/guanidinium receptive sites (IGRS) or imidazoline receptors in the eel intestine. Competition experiments revealed that rank order to displace 2 nM [3H]clonidine from their binding sites was as follows: guanabenz &gt
cirazoline = naphazoline = UK14304 = ST587 ≥ clonidine ≥ idazoxan = RX821002 = tolazoline &gt
ST93 = oxymetazoline = amiloride = ST91 &gt
yohimbine = efaroxan = rauwolscine ≥ adrenaline = ST567 = histamine = agmatine. The rank order was different from those in I1 or I2 sites of IGRS reported in various mammalian tissues, suggesting existence of new IGRS, non I1 and non I2 sites, in the eel intestine. In addition, structure-affinity relationships are discussed from the results of competition experiments. Although physiological role of IGRS is not clear yet even in mammalian cells/tissues, eel intestine may be a good model to elucidate how the IGRS act in the cell and to decide what is the endogenous ligand for the IGRS, since eel intestine contains great amount of IGRS and it responds to guanabenz, an exogenous clonidine derivative.

DOI： 10.2108/zsj.15.205

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DOI： 10.2108/zsj.15.205

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記述言語：英語  

We have examined several physiological variables related to salinity acclimation in the euryhaline tilapia, Oreochromis mossambicus. Tilapia reared in fresh water (FW) were transferred to FW, isosmotic salinity (ISO, 12‰%) and 75% seawater (SW, 25‰). Oxygen consumption, plasma levels of cortisol, growth hormone (GH), prolactins (tPRL177 and tPRL188), glucose, ions (Na+, K+, Cl-), and gill Na+,K+-ATPase activities were measured for up to 4 days in each salinity treatment. Plasma Na+ and Cl- concentrations were elevated 1 day after transfer to SW, but returned to FW values on day 4. Plasma cortisol and glucose levels were higher in FW and ISO than in SW 1 day after transfer. Plasma GH levels of tilapia in SW increased above FW and ISO values after 4 days, whereas plasma PRL levels decreased in ISO and SW compared to FW at 4 days. These results are consistent with the possible osmoregulation roles of GH and PRL in SW and FW, respectively. Gill Na+,K+-ATPase activity of tilapia in SW increased more than 2-fold over the FW value after 4 days, but activity of this enzyme did not change in ISO. Oxygen consumption rates of tilapia in SW were significantly elevated 4 days after transfer compared to FW and ISO. The results of this study indicate that the physiological changes associated with SW acclimation in tilapia represents a significant short-term energetic cost, and may account for as much as 20% of total body metabolism after 4 days in SW.

DOI： 10.1016/S0300-9629(96)00261-7

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記述言語：英語  

DOI： 10.1016/S0300-9629(96)00261-7

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DOI： 10.1073/pnas.94.5.2068

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DOI： 10.1073/pnas.94.5.2068

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DOI： 10.1016/0014-5793(96)00172-X

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Cold acclimation of rainbow trout cells is considered to be mediated by alterations in the mRNAs and proteins present in cold-treated cells. A subtracted cDNA library from cold-treated rainbow trout RTG-2 cells was constructed and screened to isolate cDNA induced in the cold-treated cells in order to elucidate which genes are induced by cold acclimation. A set of cDNA clones encoding three members of ferritin H isoforms was isolated as cold- inducible genes. Northern blot analysis and nuclear run-on transcription assay showed that the transcription and accumulation of the ferritin H isoforms mRNA were enhanced by cold acclimation. Furthermore, the ferritin level in the trout cells increased on cold acclimation in response to a temperature shift from 22 °C to 4 °C. When the trout cells were subjected to 4 °C under the condition of a decreased ferritin H level obtained by the addition of an antisense oligonucleotide, cell growth was apparently inhibited. These findings indicate an association between the induction of ferritin H and cellular mechanisms during cold acclimation of trout cells.

DOI： 10.1074/jbc.271.43.26908

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DOI： 10.1073/pnas.90.5.1912

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DOI： 10.1073/pnas.90.5.1912

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