MISC - SAKAMOTO Hirotaka

オキシトシン放出を調節する1回膜貫通型タンパク質CD38のはたらき

坂本浩隆, 越智拓海, 越智拓海, 大坪秋人, 川上奈津子, 坂本竜哉

日本解剖学会総会・全国学術集会講演プログラム・抄録集   127th (CD-ROM)   2022

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ガストリン放出ペプチド神経系による体性感覚調節機構

高浪景子, 坂本浩隆

日本解剖学会総会・全国学術集会講演プログラム・抄録集   127th (CD-ROM)   2022

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オキシトシンによる脊髄射精中枢を介した男性性機能の調節メカニズム

越智拓海, 越智拓海, 坂本竜哉, 坂本浩隆

日本性機能学会中部総会プログラム・抄録集   32nd   2022

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バソプレシンニューロンにおいて異常タンパク凝集体は小胞体から輸送隔離されることなく小胞体内部で分解される-家族性中枢性尿崩症モデルマウスを用いた検討-

宮田崇, 萩原大輔, 津村哲郎, 蓬臺優一, 川口頌平, 栗本隼樹, 高木博史, 須賀英隆, 川上奈津子, 坂本浩隆, 松本真実, 大野伸彦, 大野伸彦, 有馬寛

日本内分泌学会雑誌   97 ( 1 )   2021

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内側視索前野の性的二型核に発現するガストリン放出ペプチド系は雄ラットの性的活性を制御する

前嶋翔, 野村黎, 高浪景子, 坂本竜哉, 坂本浩隆

日本神経内分泌学会学術集会プログラム・抄録集   47th   2021

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17β-エストラジオールによる痒み閾値調節

高浪景子, 歌大介, 松田賢一, 河田光博, 河田光博, CARSTENS Earl, 坂本竜哉, 坂本浩隆

日本神経内分泌学会学術集会プログラム・抄録集   47th   2021

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コペプチン動態に着目したバソプレシン前駆体・プロセシング機構の解明

大坪秋人, 川上奈津子, 前嶋翔, 坂本竜哉, 坂本浩隆

バゾプレシン研究会プログラム・講演抄録   31st   2021

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Localization of Oxytocin and Oxytocin Receptors in the Rat Testis

ZUN Miho, 伊藤隆志, 坂本竜哉, 小谷享, 坂本浩隆, 越智拓海, 越智拓海

Science Journal of Kanagawa University   32   2021

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痒みの神経伝達基盤-疾患モデル動物を用いた検討-

高浪景子, 小出剛, 坂本浩隆

日本内分泌学会雑誌   97 ( 1 )   2021

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性経験のないオスメダカは初めての性的パートナーに対して配偶者選好性を示す

大門将寛, 安齋賢, 勝村啓史, 坂本浩隆, 竹内秀明

日本動物行動学会大会発表要旨集   40th (CD-ROM)   2021

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ミネラルコルチコイド受容体ノックアウト(MR-KO)メダカにおける異常な視覚依存行動の観察

唐嘉榮, 今野紀文, 中町智哉, 坂本浩隆, 坂本竜哉, 松田恒平

日本比較内分泌学会大会及びシンポジウムプログラム・講演要旨   45th   2021

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扁形動物ヒラムシから紐解くGnRHの祖先型機能

森俊輔, 酒井丈実, 濱田麻友子, 坂本竜哉, 坂本浩隆

日本比較内分泌学会大会及びシンポジウムプログラム・講演要旨   45th   2021

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雄ラット扁桃体内側核後背側部におけるガストリン放出ペプチド系を介した性行動調節メカニズムの解明

大坪秋人, 上田涼太, 高松廉, 大野智輝, 前嶋翔, 越智拓海, 越智拓海, 犬束歩, 尾仲達史, 坂本竜哉, 坂本浩隆

日本神経内分泌学会学術集会プログラム・抄録集   47th   2021

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雄ラット視床下部腹内側核オキシトシン受容体ニューロン系による食欲と性欲のスイッチング機構

立石沙也加, 上田涼太, 永渕詢大, 越智拓海, 越智拓海, 犬束歩, 尾仲達史, VALERY Grinevich, 坂本竜哉, 坂本浩隆

日本神経内分泌学会学術集会プログラム・抄録集   47th   2021

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'Central' Actions of Corticosteroid Signaling Suggested by Constitutive Knockout of Corticosteroid Receptors in Small Fish. International journal

Tatsuya Sakamoto, Hirotaka Sakamoto

Nutrients   11 ( 3 )   2019.3

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This review highlights recent studies of the functional implications of corticosteroids in some important behaviors of model fish, which are also relevant to human nutrition homeostasis. The primary actions of corticosteroids are mediated by glucocorticoid receptor (GR) and mineralocorticoid receptor (MR), which are transcription factors. Zebrafish and medaka models of GR- and MR-knockout are the first constitutive corticosteroid receptor-knockout animals that are viable in adulthood. Similar receptor knockouts in mice are lethal. In this review, we describe the physiological and behavioral changes following disruption of the corticosteroid receptors in these models. The GR null model has peripheral changes in nutrition metabolism that do not occur in a mutant harboring a point mutation in the GR DNA-binding domain. This suggests that these are not "intrinsic" activities of GR. On the other hand, we propose that integration of visual responses and brain behavior by corticosteroid receptors is a possible "intrinsic"/principal function potentially conserved in vertebrates.

DOI： 10.3390/nu11030611

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雄ラットにおける性行動への内側視索前野の性的二型核の関与

前嶋翔, 野村黎, 高浪景子, 高浪景子, 坂本竜哉, 坂本浩隆

日本比較内分泌学会大会及びシンポジウムプログラム・講演要旨   44th   2019

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魚類をモデルとしたアンドロゲンによる二次性徴形質の発現と多様化の分子機構

荻野由紀子, 安齋賢, 坂本浩隆, 渡辺英治, 成瀬清, 井口泰泉

日本分子生物学会年会プログラム・要旨集(Web)   42nd   2019

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Early-life exposure to Tris(1,3-dichloroisopropyl) phosphate induces dose-dependent suppression of sexual behavior in male rats. International journal

Manami Kamishima, Tatsuya Hattori, Go Suzuki, Hidenori Matsukami, Chiaki Komine, Yasuyuki Horii, Gen Watanabe, Takumi Oti, Hirotaka Sakamoto, Tomoko Soga, Ishwar S Parhar, Yasuhiko Kondo, Hidetaka Takigami, Maiko Kawaguchi

Journal of applied toxicology : JAT   38 ( 5 )   649 - 655   2018.5

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Exposure to endocrine-disrupting chemicals may adversely affect animals, particularly during development. Tris(1,3-dichloroisopropyl) phosphate (TDCIPP) is an organophosphate with anti-androgen function in vitro that is present in indoor dust at relatively high concentrations. In male rats, androgens are necessary for the development of reproductive organs, as well as the endocrine and central nervous systems. However, we currently do not know the exact effects of TDCIPP exposure through suckling on subsequent reproductive behavior in males. Here, we show that TDCIPP exposure (25-250 mg kg-1 via oral administration over 28 consecutive days post-birth) suppressed male sexual behavior and reduced testes size. These changes were dose-dependent and appeared first in adults rather than in juveniles. These results demonstrate that TDCIPP exposure led to normal body growth and appearance in juveniles, but disrupted the endocrine system and physiology in adults. Therefore, assays should be performed using adult animals to ensure accuracy, and to confirm the influence of chemical substances given during early mammalian life.

DOI： 10.1002/jat.3569

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Other Link： http://orcid.org/0000-0002-6224-4591

Analyzing the effects of co-expression of chick (Gallus gallus) melanocortin receptors with either chick MRAP1 or MRAP2 in CHO cells on sensitivity to ACTH(1–24) or ACTH(1–13)NH2: Implications for the avian HPA axis and avian melanocortin circuits in the hypothalamus

Alexa L. Thomas, Fumihiko Maekawa, Takaharu Kawashima, Hirotaka Sakamoto, Tatsuya Sakamoto, Perry Davis, Robert M. Dores

General and Comparative Endocrinology   256   50 - 56   2018.1

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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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ラット内側視索前野において雄の性行動を司る神経回路系の解析

野村黎, 高浪景子, 高浪景子, 坂本竜哉, 坂本浩隆

日本神経内分泌学会学術集会プログラム・抄録集   45th   2018

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小胞体内凝集体形成機序の解明-家族性中枢性尿崩症モデルマウスを用いた検討-

宮田崇, 萩原大輔, 萩原大輔, 橡谷昌佳, 森下啓明, 坂本浩隆, 有馬寛

日本内分泌学会雑誌   94 ( 1 )   2018

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マウスとラットを用いた痒み伝達機構の比較解析

高浪景子, 坂本浩隆, 小出剛

日本分子生物学会年会プログラム・要旨集(Web)   41st   2018

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バソプレシンニューロンにおける異常蛋白の処理機構に小胞体シャペロンBiPおよびライソソームが関与する

宮田崇, 萩原大輔, 萩原大輔, 川口頌平, 栗本隼樹, 尾崎創, 光本一樹, 高木博史, 須賀英隆, 坂本浩隆, 有馬寛

バゾプレシン研究会プログラム・講演抄録   29th   2018

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霊長類ニホンザルにおけるGastrin-releasing peptide神経系の機能局在

高浪景子, 伊藤隆志, 越智拓海, 小林靖尚, 佐藤慧太, 上田康雅, 坂本竜哉, 坂本浩隆

日本解剖学会総会・全国学術集会講演プログラム・抄録集   123rd   2018

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Identification of the sexually dimorphic gastrin-releasing peptide system in the lumbosacral spinal cord that controls male reproductive function in the mouse and Asian house musk shrew (Suncus murinus)

Kei Tamura, Yasuhisa Kobayashi, Asuka Hirooka, Keiko Takanami, Takumi Oti, Takamichi Jogahara, Sen-ichi Oda, Tatsuya Sakamoto, Hirotaka Sakamoto

JOURNAL OF COMPARATIVE NEUROLOGY   525 ( 7 )   1586 - 1598   2017.5

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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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蛍光タンパク質Venus導入ラットを用いた雄の性機能を司る脊髄神経回路系の解析

越智拓海, 高浪景子, 松田賢一, 河田光博, 坂本竜哉, 坂本浩隆

日本アンドロロジー学会総会記事   36th   2017

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コペプチンからバソプレシン遺伝子の転写-翻訳-プロセシングを解析する試み

坂本浩隆, 坂本浩隆, 佐藤慧太, 高浪景子, 大坪秋人, 坂本竜哉, MORRIS John F.

日本神経内分泌学会学術集会プログラム・抄録集   44th   2017

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行動を司る時間・空間的神経内分泌制御メカニズム

坂本浩隆

日本神経内分泌学会学術集会プログラム・抄録集   43rd   2016

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雄の性機能を司る脊髄神経回路系の機能解析:Grp-promoter-Venusトランスジェニックラットの作出と利用

越智拓海, 高浪景子, 高浪景子, 松田賢一, 河田光博, 坂本竜哉, 坂本浩隆

日本神経内分泌学会学術集会プログラム・抄録集   43rd   2016

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ニホンザル脊髄における痒み特異的伝達分子gastrin-releasing peptide受容体の発現

伊藤隆志, 高浪景子, 高浪景子, 越智拓海, 小林靖尚, 小林靖尚, 佐藤慧太, 高橋俊次, 上田康雅, 坂本竜哉, 坂本浩隆

日本組織細胞化学会総会・学術集会講演プログラム・予稿集   57th   2016

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「基礎系」オキシトシンと性機能

坂本浩隆

日本性機能学会雑誌   31 ( 2 )   2016

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Requirement of vasotocin for being a winner in love triangle relationships (two males and one female) of medaka fish Invited

Saori Yokoi, Tatsuya Sakamoto, Hirotaka Sakamoto, Hideaki Takeuchi

37 ( 6 )   591 - 597   2015.12

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)  

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超高圧電子顕微鏡・トモグラフィー法を用いた脊髄内痒み神経ネットワークの解析

坂本浩隆, 佐藤慧太, 高浪景子, 高浪景子, 高浪景子, 村田和義, 河田光博, 坂本竜哉

日本内分泌学会雑誌   91 ( 2 )   525 - 525   2015.9

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In vivo imaging of axonal transport of mitochondria in the diseased and aged mammalian CNS

Yuji Takihara, Masaru Inatani, Kei Eto, Toshihiro Inoue, Alexander Kreymerman, Seiji Miyake, Shinji Ueno, Masatoshi Nagaya, Ayami Nakanishi, Keiichiro Iwao, Yoshihiro Takamura, Hirotaka Sakamoto, Keita Satoh, Mineo Kondo, Tatsuya Sakamoto, Jeffrey L. Goldberg, Junichi Nabekura, Hidenobu Tanihara

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   112 ( 33 )   10515 - 10520   2015.8

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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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P4-10 医学・生物学分野における超高圧電子顕微鏡・トモグラフィー法の再考 : A revisiting study for 3D-EM(細胞生物・電子顕微鏡,ポスター発表,組織化学のモーダルシフト,第56回日本組織細胞化学会総会・学術集会)

坂本 浩隆, 佐藤 慧太, 高浪 景子, 村田 和義, 河田 光博, 坂本 竜哉

日本組織細胞化学会総会プログラムおよび抄録集   ( 56 )   71 - 71   2015

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P9-01 哺乳類三叉神経知覚系の比較組織学的解析(感覚・皮膚・歯,ポスター発表,組織化学のモーダルシフト,第56回日本組織細胞化学会総会・学術集会)

高浪 景子, 向井 啓起, 井上 海平, 田村 圭, 越智 拓海, 上田 康雅, 河田 光博, 坂本 竜哉, 坂本 浩隆

日本組織細胞化学会総会プログラムおよび抄録集   ( 56 )   80 - 80   2015

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Other Link： http://search.jamas.or.jp/link/ui/2016046996

P8-03 霊長類ニホンザルの脊髄におけるgastrin-releasing peptide系の存在(脳機能,ポスター発表,組織化学のモーダルシフト,第56回日本組織細胞化学会総会・学術集会)

伊藤 隆志, 高浪 景子, 越智 拓海, 小林 靖尚, 佐藤 慧太, 高橋 俊次, 上田 康雅, 坂本 竜哉, 坂本 浩隆

日本組織細胞化学会総会プログラムおよび抄録集   ( 56 )   78 - 78   2015

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Other Link： http://search.jamas.or.jp/link/ui/2016046985

P4-12 AVP-eGFP TgラットにおけるインビボGFP動態(細胞生物・電子顕微鏡,ポスター発表,組織化学のモーダルシフト,第56回日本組織細胞化学会総会・学術集会)

佐藤 慧太, 越智 拓海, 加藤 明子, 上田 陽一, Morris John F, 坂本 竜哉, 坂本 浩隆

日本組織細胞化学会総会プログラムおよび抄録集   ( 56 )   72 - 72   2015

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Other Link： http://search.jamas.or.jp/link/ui/2016046963

かゆみを伝える神経ネットワーク

高浪景子, 坂本浩隆, 宮崎直幸, 村田和義, 佐藤慧太, 坂本竜哉, 谷田任司, 山田俊児, 松田賢一, 河田光博

日本内分泌学会雑誌   90 ( 2 )   602 - 602   2014.9

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Bioimaging histochemistry 免疫組織化学法を用いた体性感覚ニューロン系の3次元微細構造解析

高浪 景子, 坂本 浩隆, 宮崎 直幸, 村田 和義, 佐藤 慧太, 坂本 竜哉, 河田 光博

日本組織細胞化学会総会・学術集会講演プログラム・予稿集   55回   65 - 65   2014.9

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Quality Control of Photosystem II: Direct Imaging of the Changes in the Thylakoid Structure and Distribution of FtsH Proteases in Spinach Chloroplasts under Light Stress

Miho Yoshioka-Nishimura, Daisuke Nanba, Takashi Takaki, Chikako Ohba, Nodoka Tsumura, Noriko Morita, Hirotaka Sakamoto, Kazuyoshi Murata, Yasusi Yamamoto

PLANT AND CELL PHYSIOLOGY   55 ( 7 )   1255 - 1265   2014.7

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Under light stress, the reaction center-binding protein D1 of PSII is photo-oxidatively damaged and removed from PSII complexes by proteases located in the chloroplast. A protease considered to be responsible for degradation of the damaged D1 protein is the metalloprotease FtsH. We showed previously that the active hexameric FtsH protease is abundant at the grana margin and the grana end membranes, and this homo-complex removes the photodamaged D1 protein in the grana. Here, we showed a change in the distribution of FtsH in spinach thylakoids during excessive illumination by transmission electron microscopy (TEM) and immunogold labeling of FtsH. The change in distribution of the protease was accompanied by structural changes to the thylakoids, which we detected using spinach leaves by TEM after chemical fixation of the samples. Quantitative analyses showed several characteristic changes in the structure of the thylakoids, including shrinkage of the grana, outward bending of the marginal portions of the thylakoids and an increase in the height of the grana stacks under excessive illumination. The increase in the height of the grana stacks may include swelling of the thylakoids and an increase in the partition gaps between the thylakoids. These data strongly suggest that excessive illumination induces partial unstacking of the thylakoids, which enables FtsH to access easily the photodamaged D1 protein. Finally three-dimensional tomography of the grana was recorded to observe the effect of light stress on the overall structure of the thylakoids.

DOI： 10.1093/pcp/pcu079

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Distribution of Gastrin-Releasing Peptide in the Rat Trigeminal and Spinal Somatosensory Systems

Keiko Takanami, Hirotaka Sakamoto, Ken Ichi Matsuda, Keita Satoh, Takashi Tanida, Shunji Yamada, Kaihei Inoue, Takumi Oti, Tatsuya Sakamoto, Mitsuhiro Kawata

JOURNAL OF COMPARATIVE NEUROLOGY   522 ( 8 )   1858 - 1873   2014.6

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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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光化学系IIのquality control:光ストレス下でのホウレンソウ葉緑体のグラナ膜構造の変化

難波大介, 西村美保, 坂本浩隆, 村田和義, 高木孝士, 山本泰

日本植物生理学会年会要旨集   55th   328   2014.3

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Identification of CNS Neurons Innervating the Levator Ani and Ventral Bulbospongiosus Muscles in Male Rats

Amy D. Dobberfuhl, Takumi Oti, Hirotaka Sakamoto, Lesley Marson

JOURNAL OF SEXUAL MEDICINE   11 ( 3 )   664 - 677   2014.3

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IntroductionThe pelvic striated muscles play an important role in mediating erections and ejaculation, and together these muscles compose a tightly coordinated neuromuscular system that is androgen sensitive and sexually dimorphic.
AimTo identify spinal and brains neurons involved in the control of the levator ani (LA) and bulbospongiosus (BS) in the male adult and preadolescent rat.
MethodsRats were anesthetized, and the transsynaptic retrograde tracer pseudorabies virus (PRV) was injected into the LA muscle of adults or the ventral BS muscle in 30-day-old rats. After 3-5 days rats were sacrificed, and PRV-labeled neurons in the spinal cords and brains were identified using immunohistochemistry. The presence of gastrin-releasing peptide (GRP) in the lumbar spinal neurons was examined.
Main Outcomes MeasuresThe location and number of PRV-labeled neurons in the spinal cord and brain and GRP colocalization in the lumbar spinal cord.
ResultsPRV-labeled spinal interneurons were found distributed throughout T11-S1 of the spinal cord, subsequent to dorsal medial motoneuron infection. The majority of spinal interneurons were found in the lumbosacral spinal cord in the region of the dorsal gray commissure and parasympathetic preganglionic neurons. Preadolescent rats had more PRV-labeled spinal interneurons at L5-S1 where the motoneurons were located but relatively less spread rostrally in the spinal cord compared with adults. Lumbar spinothalmic neurons in medial gray of L3-L4 co-localized PRV and GRP. In the brain consistent labeling was seen in areas known to be involved in male sexual behavior including the ventrolateral medulla, hypothalamic paraventricular nucleus, and medial preoptic area.
ConclusionCommon spinal and brain pathways project to the LA and BS muscles in the rat suggesting that these muscles act together to coordinate male sexual reflexes. Differences may exist in the amount of synaptic connections/neuronal pathways in adolescents compared with adults. Dobberfuhl AD, Oti T, Sakamoto H, and Marson L. Identification of CNS neurons innervating the levator ani and ventral bulbospongiosus muscles in male rats. J Sex Med 2014;11:664-677.

DOI： 10.1111/jsm.12418

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W3-02 免疫組織化学法を用いた体性感覚ニューロン系の3次元微細構造解析(W3 Bioimaging histochemistry,ワークショップ,第55回日本組織細胞化学会総会・学術集会 第11回日中合同組織細胞化学セミナー)

高浪 景子, 坂本 浩隆, 宮崎 直幸, 村田 和義, 佐藤 慧太, 坂本 竜哉, 河田 光博

日本組織細胞化学会総会プログラムおよび抄録集   ( 55 )   65 - 65   2014

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光化学系IIのquality control:光ストレス下でのホウレンソウ葉緑体チラコイド膜構造変化の電子顕微鏡による観察

難波大介, 西村美保, 坂本浩隆, 村田和義, 高木孝士, 山本泰

日本植物学会大会研究発表記録   77th   231   2013.8

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光化学系IIのquality control:電子顕微鏡を用いた光ストレス下のホウレンソウ葉緑体のグラナチラコイド膜構造の観察

難波大介, 西村美保, 大庭千加子, 坂本浩隆, 村田和義, 高木孝士, 山本泰

日本植物生理学会年会要旨集   54th   238   2013.3

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Potential roles of arginine-vasotocin in the regulation of aggressive behavior in the mudskipper (Periophthalmus modestus)

N. Kagawa

General and Comparative Endocrinology   194   257 - 263   2013

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DOI： 10.1016/j.ygcen.2013.09.023.

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Potential roles of arginine-vasotocin in the regulation of aggressive behavior in the mudskipper (Periophthalmus modestus)

N. Kagawa

General and Comparative Endocrinology   194   257 - 263   2013

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DOI： 10.1016/j.ygcen.2013.09.023.

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超高圧電子顕微鏡を用いたラット脊髄gastrin‐releasing peptide系を中心とした脊髄内局所神経回路の三次元的解析

越智拓海, 佐藤慧太, 齊藤和裕, 村田和義, 河田光博, 坂本竜哉, 坂本浩隆

日本解剖学会総会・全国学術集会講演プログラム・抄録集   118th   167   2013

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Three-dimensional evaluation of the spinal local neural network revealed by the high-voltage electron microscopy: a double immunohistochemical study

Takumi Oti, Keita Satoh, Kazuhiro Saito, Kazuyoshi Murata, Mitsuhiro Kawata, Tatsuya Sakamoto, Hirotaka Sakamoto

HISTOCHEMISTRY AND CELL BIOLOGY   138 ( 4 )   693 - 697   2012.10

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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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光化学系IIのquality control:電子顕微鏡を用いた光ストレス下のホウレンソウ葉緑体のグラナチラコイド膜構造の観察

難波大介, 西村(吉岡)美保, 坂本浩隆, 村田和義, 山本泰

日本植物学会大会研究発表記録   76th   222   2012.9

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Brain-spinal cord neural circuits controlling male sexual function and behavior

Hirotaka Sakamoto

Neuroscience Research   72 ( 2 )   103 - 116   2012.2

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Men and women exhibit differences in sexual behavior. This indicates that neural circuits within the central nervous system (CNS) that control sexual behavior differ between the sexes, although differences in behavior are also influenced by sociocultural and hormonal factors. Sexual differentiation of the body and brain occurs during the embryonic and neonatal periods in humans and persists into adulthood with relatively low plasticity. Male sexual behavior is complex and depends on intrinsic and extrinsic factors, including olfactory, somatosensory and visceral cues. Many advances in our understanding of sexually dimorphic neural circuits have been achieved in animal models, but major issues are yet to be resolved. This review summarizes the sexually dimorphic nuclei controlling male sexual function in the rodent CNS and focuses on the interactions of the brain-spinal cord neural networks controlling male sexual behavior. Possible factors that relate findings from animal studies to human behavior are also discussed. © 2011 Elsevier Ireland Ltd and the Japan Neuroscience Society.

DOI： 10.1016/j.neures.2011.11.002

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Achieve orgasm? Oxytocin triggers ejaculation in men

H. Sakamoto

Reproductive System & Sexual Disorders   1   e101-e101   2012

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Hemolymph osmotic, ionic status, and branchial Na⁺/K⁺-ATPase activity under varying environmental conditions in the intertidal grapsid crab, Gaetice depressusd

Takeshi Nanba, Hideya Takahashi, Tsukasa Abe, Waichirou Godo, Maho Ogoshi, Hirotaka Sakamoto, Naoaki Tsutsui, Tatsuya Sakamoto

International Aquatic Research   4 ( 1 )   1 - 12   2012

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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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Immunohistochemical Analysis of the Effects of Estrogen on Intraarticular Neurogenic Inflammation in a Rat Anterior Cruciate Ligament Transection Model of Osteoarthritis

Atsuhiko Yoshida, Toru Morihara, Ken-ichi Matsuda, Hirotaka Sakamoto, Yuji Arai, Yoshikazu Kida, Mitsuhiro Kawata, Toshikazu Kubo

CONNECTIVE TISSUE RESEARCH   53 ( 3 )   197 - 206   2012

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Synovitis is considered as one of the factors associated with the pathogenesis of osteoarthritis (OA). There is currently a significant amount of research linking estrogen deficiencies with the development of OA in estrogen-deficient women, including postmenopausal women; however, the exact etiology remains unclear. Various neuropeptides, such as substance P (SP) and calcitonin gene-related peptide (CGRP), have been shown to contribute to synovitis in OA joints, and the influence of estrogen on the expressions of SP and CGRP in the synovium of OA joints has been noted. After ovariectomy (OVX) followed by estradiol (E2) replacement, 24 female rats were divided into three groups: OVX group, OVX + E2 replacement group (E2 group), and a sham group. All rats underwent transection of the anterior cruciate ligament at the same time. After 30 days, the histological findings of knee joints by hematoxylin-eosin staining and immunofluorescence staining of protein gene product 9.5 (pan-neuronal marker), SP, and CGRP were compared among experimental groups. The degree of synovitis in the OVX group was higher than in the E2 and sham groups. No significant differences in the density of protein gene product 9.5-immunoreactive nerve fibers were observed among the three experimental groups, but the density of SP- or CGRP-immunoreactive nerve fibers in the OVX group was significantly higher than in the E2 and sham groups. These findings suggest that estrogen partly regulates intraarticular neurogenic inflammation in OA joints by modulating the expressions of neuropeptides in the synovium.

DOI： 10.3109/03008207.2011.628059

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A Gastrin-Releasing Peptide (GRP) System in the Spinal Cord Controls Male Sexual Functions

SAKAMOTO H.

85 ( 3 )   113 - 114   2010.9

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Induction of the arginine vasopressin-enhanced green fluorescent protein fusion transgene in the rat locus coeruleus

Miwako Todoroki, Yoichi Ueta, Hiroaki Fujihara, Hiroki Otsubo, Minori Shibata, Hirofumi Hashimoto, Mizuki Kobayashi, Hirotaka Sakamoto, Mitsuhiro Kawata, Govindan Dayanithi, David Murphy, Hisanori Hiro, Ken Takahashi, Shoji Nagata

STRESS-THE INTERNATIONAL JOURNAL ON THE BIOLOGY OF STRESS   13 ( 4 )   281 - 292   2010.7

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We examined the effects of intracerebroventricular (i.c.v.) administration of colchicine on the expression of the arginine vasopressin (AVP)-enhanced green fluorescent protein (eGFP) fusion gene in rats. In rats administered i.c.v. vehicle ( control), eGFP fluorescence was observed in the supraoptic nucleus ( SON), the magnocellular division of the paraventricular nucleus (PVN), the suprachiasmatic nucleus (SCN), the median eminence ( ME) and the posterior pituitary. Two days after i.c.v. administration of colchicine, eGFP fluorescence was markedly increased in the SON, the magnocellular and parvocellular divisions of the PVN, the SCN, the ME and the locus coeruleus (LC). Immunohistochemical staining for eGFP confirmed the distribution of fluorescence in both groups. In the colchicines-administered groups, immunohistochemistry for tyrosine hydroxylase (TH) revealed that the eGFP fluorescence was co-localised with TH-immunoreactivity in the LC. Similarly, in situ hybridization histochemistry for eGFP mRNA revealed a significant increase in gene expression in the LC, the SON and the PVN 12-48 h after administration of colchicine. Our results indicate that the synthesis of AVP-eGFP is upregulated in noradrenergic neurones in the LC after colchicine administration. This implies that AVP and noradrenaline, originating from LC neurones, might play a role in response to chronic stress.

DOI： 10.3109/10253890903383406

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Aldosterone-Sensitive Nucleus Tractus Solitarius Neurons Regulate Sensitivity of the Baroreceptor Reflex in High Sodium-Loaded Rats

Tomoharu Masuda, Yasutoshi Hirabara, Yusuke Nakamura, Akiko Chishaki, Mai Tsuruhisa, Masayuki Miyakawa, Kenji Honda, Ryo Saito, Hirotaka Sakamoto, Mitsuhiro Kawata, Yukio Takano

JOURNAL OF PHARMACOLOGICAL SCIENCES   112 ( 4 )   482 - 486   2010.4

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We examined the role of aldosterone-sensitive neurons in the nucleus tractus solitarius (NTS) in the arterial baroreceptor reflex (baroreflex) function. Baroreflex sensitivity was induced by phenylephrine in high sodium-loaded rats and was significantly reduced. This baroreflex sensitivity was reversed by microinjection of the mineralocorticoid receptor (MR) antagonist eplerenone into the NTS. 11 beta-Hydroxysteroid dehydrogenase type 2 neurons and MR were also identified in the NTS. These data suggest that the aldosterone-sensitive neurons in the NTS may have an important role in baroreflex function.

DOI： 10.1254/jphs.09362SC

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Expression of G protein-coupled receptor 30 in the spinal somatosensory system

Keiko Takanami, Hirotaka Sakamoto, Ken-Ichi Matsuda, Koji Hosokawa, Mayumi Nishi, Eric R. Prossnitz, Mitsuhiro Kawata

BRAIN RESEARCH   1310   17 - 28   2010.1

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Estrogens were originally identified as the primary sex steroid hormones in females and regulators of reproductive function and sexual behavior, but it has long been suggested that estrogens also have local effects on the somatosensory system at the spinal cord level. it is well known that the effects of estrogens are mediated by nuclear estrogen receptors (ERs) through genomic action, but recently a membrane-bound G protein-coupled receptor, GPR30, was identified as a non-genomic estrogen receptor. in this study we investigated the presence and localization of GPR30 in the rat spinal cord and dorsal root ganglion (DRG) in comparison with ER alpha. Using immunohistochemistry and in situ hybridization, we showed the expression of GPR30 in DRG neurons in male and female rats at mRNA and protein levels without specific sexual difference. A dense accumulation of GPR30 immunoreactivity was observed in the outer layer of the spinal dorsal horn, and selective spinal dorsal rhizotomy revealed that GPR30 was transported from the DRG to terminals located in the spinal dorsal horn. GPR30 expression was downregulated in DRG neurons of ovariectomized female rats. The spinal somatosensory system might be modulated by estradiol via putative membrane ER, GPR30-mediated mechanism. (C) 2009 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.brainres.2009.11.004

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性機能とgastrin-releasing peptide (GRP) ニューロンシステム − EDの中枢性病態生理解明に向けて −

坂本 浩隆, 河田光博

自律神経   47   82 - 85   2010

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Estrogen regulate itch sensation

Keiko Takanami, Ken-ichi Matsuda, Hirotaka Sakamoto, Yukie Hirahara, Mitsuhiro Kawata

NEUROSCIENCE RESEARCH   68   E162 - E162   2010

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：ELSEVIER IRELAND LTD  

DOI： 10.1016/j.neures.2010.07.2292

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超高圧電子顕微鏡を用いた脊髄gastrin-releasing peptide (GRP) 系から球海綿体脊髄核ニューロンへのシナプス入力の可視化

坂本 浩隆

比較内分泌学   36 ( 137 )   146 - 148   2010

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Language：Japanese   Publisher：Japan Society for Comparative Endocrinology  

DOI： 10.5983/nl2008jsce.36.146

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The neurobiology of psychogenic erectile dysfunction in the spinal cord

H. Sakamoto

Journal of Andrology   21 ( 4 )   432 - 435   2010

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Characterization of In Vivo Effects of Platelet-Rich Plasma and Biodegradable Gelatin Hydrogel Microspheres on Degenerated Intervertebral Discs

Kazuhide Sawamura, Takumi Ikeda, Masateru Nagae, Shin-ichi Okamoto, Yasuo Mikami, Hitoshi Hase, Kazuya Ikoma, Tetsuya Yamada, Hirotaka Sakamoto, Ken-ichi Matsuda, Yasuhiko Tabata, Mitsuhiro Kawata, Toshikazu Kubo

TISSUE ENGINEERING PART A   15 ( 12 )   3719 - 3727   2009.12

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Language：English   Publisher：MARY ANN LIEBERT, INC  

We have previously shown that administration of platelet-rich plasma-impregnated gelatin hydrogel microspheres (PRP-GHMs) into a degenerated intervertebral disc (IVD) markedly suppresses progression of IVD degeneration. In the current study, we characterized the in vivo effects of PRP-GHM treatment in a degenerated IVD model in rabbit. On magnetic resonance images, the IVD height was significantly greater after treatment with PRP-GHMs compared with phosphate-buffered saline-impregnated GHMs, PRP without GHMs, and needle puncture only. Water content was also preserved in PRP-GHM-treated IVDs. Consistent with this observation, the mRNA expression of proteoglycan core protein and type II collagen was significantly higher after PRP-GHM treatment compared with other treatment groups. No proliferating cells were found in the nucleus pulposus and inner annulus fibrosus in any groups, but the number of apoptotic cells in the nucleus pulposus after PRP-GHM treatment was significantly lower than that after other treatments. These results provide an improved understanding of the therapeutic effects of PRP-GHM treatment of degenerated IVDs.

DOI： 10.1089/ten.tea.2008.0697

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Androgen Regulates the Sexually Dimorphic Gastrin-Releasing Peptide System in the Lumbar Spinal Cord that Mediates Male Sexual Function (vol 150, pg 3672, 2009)

Hirotaka Sakamoto, Keiko Takanami, Damian G. Zuloaga, Kenichi Matsuda, Cynthia L. Jordan, S. Marc Breedlove, Mitsuhiro Kawata

ENDOCRINOLOGY   150 ( 9 )   4461 - 4461   2009.9

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Gastrin-Releasing Peptide System in the Spinal Cord Controls Male Sexual Behaviour

H. Sakamoto, M. Kawata

JOURNAL OF NEUROENDOCRINOLOGY   21 ( 4 )   432 - 435   2009.4

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Language：English   Publishing type：Book review, literature introduction, etc.   Publisher：WILEY-BLACKWELL PUBLISHING, INC  

The lumbar spinal cord contains local neural circuits that are important in regulating male sexual behaviours, but the molecular mechanisms underlying these systems remain elusive. Gastrin-releasing peptide (GRP) is a member of the bombesin-like peptide family first isolated from the porcine stomach. Despite extensive pharmacological studies on the activity of bombesin-like peptides administered to mammals, little is known about the physiological functions of GRP in the spinal cord. We review recent findings on a system of neurones in the upper lumbar spinal cord, within the recently reported ejaculation generator, projecting axons containing GRP to the lower lumbar spinal cord and innervating regions known to control erection and ejaculation.

DOI： 10.1111/j.1365-2826.2009.01847.x

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In Vivo Effects of Ovarian Steroid Hormones on the Expressions of Estrogen Receptors and the Composition of Extracellular Matrix in the Anterior Cruciate Ligament in Rats

Atsuhiko Yoshida, Toru Morihara, Yoshiteru Kajikawa, Yuji Arai, Yasushi Oshima, Toshikazu Kubo, Ken-ichi Matsuda, Hirotaka Sakamoto, Mitsuhiro Kawata

CONNECTIVE TISSUE RESEARCH   50 ( 2 )   121 - 131   2009

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Female athletes have a significantly higher rate of anterior cruciate ligament (ACL) injury than their male counterparts. Sex steroid hormones are considered to have an influence as risk factors for female ACL injuries. We hypothesized that estrogen and progesterone have specific and synergistic influences on the composition of extracellular matrix in ACL. By ovariectomy (OVX) followed by subcutaneous estradiol (E2) and/or progesterone (P4) replacement, 40 female rats were divided into 5 groups: E2, P4, combined E2 and P4 (EP), OVX control, and sham group. After 30 days, using undecalcified sections of knee joints in conjunction with immunofluorescence staining of estrogen receptor and (ER and ER), collagen types 1 and 3, and cartilage oligomeric matrix protein (COMP), the immunoreactivities of these proteins in two distinct parts of ACL, proximal and middle portions, were compared semiquantitatively among experimental groups. By E2 replacement, the expressions of ER in ACL fibroblasts were elevated compared to the OVX group. At the proximal portion, the immunoreactivities of type 1 collagen by E2 replacement, type 3 collagen by P4 replacement, and COMP by E2 or P4 replacement were significantly reduced. At the middle portion, the immunoreactivity of type 3 collagen was significantly elevated by E2 replacement. However, no differences were observed between the sham and OVX groups. These findings suggest that ACL is ER-dependent and that ovarian hormones alter ligament tissue composition, especially at the proximal portion. Female hormonal influences are partly involved in the biological properties of ACL.

DOI： 10.1080/03008200802531287

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Expression and its hormonal regulation of G protein-coupled receptor 30 in the rat spinal somatosensory system

Keiko Takanami, Hirotaka Sakamoto, Ken-ichi Matsuda, Koji Hosokawa, Mayumi Nishi, Eric R. Prossnitz, Mitsuhiro Kawata

NEUROSCIENCE RESEARCH   65   S106 - S106   2009

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：ELSEVIER IRELAND LTD  

DOI： 10.1016/j.neures.2009.09.474

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新規に見出した雄の性機能を脊髄レベルで制御するgastrin-releasing peptide（GRP） ニューロンシステム

坂本 浩隆

比較内分泌学   35 ( 135 )   274 - 279   2009

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Language：Japanese   Publisher：Japan Society for Comparative Endocrinology  

DOI： 10.5983/nl2008jsce.35.274

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Sexually dimorphic gastrin-releasing peptide system in the lumbar spinal cord controls masculine reproductive functions in rats

Hirotaka Sakamoto, Ken-Ichi Matsuda, Damian G. Zuloaga, Hisayuki Hongu, Etsuko Wada, Keiji Wada, Cynthia L. Jordan, S. Marc Breedlove, Mitsuhiro Kawata

NEUROSCIENCE RESEARCH   65   S41 - S41   2009

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：ELSEVIER IRELAND LTD  

DOI： 10.1016/j.neures.2009.09.047

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A high-sodium diet in streptozotocin (STZ)-induced diabetic rats impairs baroreceptor reflex; aldosterone sensitive neurons in the nucleus tractus solitarius (NTS)

Tomoharu Masuda, Yusuke Nakamura, Masayuki Miyakawa, Yasutoshi Hirabara, Kenji Honda, Ryo Saito, Kazuhiko Arimori, Hiroyuki Nakagawa, Hirotaka Sakamoto, Mitsuhiro Kawata, Yukio Takano

JOURNAL OF PHARMACOLOGICAL SCIENCES   109   271P - 271P   2009

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：JAPANESE PHARMACOLOGICAL SOC  

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バゾプレッシン(AVP)-eGFPトランスジェニックラットにおける青斑核での特異的なAVP-eGFPの合成

轟美和子, 上田陽一, 藤原広明, 大坪弘樹, 柴田美雅, 橋本弘史, 小林瑞, 坂本浩隆, 河田光博

日本病態生理学会雑誌   18 ( 2 )   2009

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REGIONAL DISTRIBUTION OF importin SUBTYPE mRNA EXPRESSION IN THE NERVOUS SYSTEM: STUDY OF EARLY POSTNATAL AND ADULT MOUSE

K. Hosokawa, M. Nishi, H. Sakamoto, Y. Tanaka, M. Kawata

NEUROSCIENCE   157 ( 4 )   864 - 877   2008.12

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Importin-alpha and beta 1 mediate the translocation of macromolecules bearing nuclear localization signals across the nuclear pore complex. Five importin-alpha isoforms have been identified in mice and six in human. Some of these importins play an important role in neural activity such as long term potentiation, but the functional differences of each isoform in the CNS are still unclear. We performed in situ hybridization (ISH) using non-isotopic probes to clarify the expression patterns of importin-alpha subtypes (alpha 5, alpha 7, alpha 1, alpha 4, alpha 3) and importin-beta 1 in the mouse CNS of adult and early postnatal stages. The mRNAs of the importin-alpha subtypes and importin 01 were expressed throughout the CNS with specific patterns; importin-alpha 5, alpha 7, alpha 3, and beta 1 showed moderate to high expression levels throughout the brain and spinal cord; importin-alpha 4 showed a lack of expression in limited regions; and importin-alpha 1 showed a low expression level throughout the brain and spinal cord but with a moderate expression level in the olfactory bulb and reticular system. We also demonstrated that importin-alpha s and beta 1 mRNAs were predominantly expressed in neurons in the adult mouse brain by using double-labeling fluorescence ISH and immunohistochemistry. Moreover, importin-alpha s and beta 1 mRNAs were detected throughout the CNS of postnatal mice and were highly expressed in the external granule layer of the cerebellar cortex on postnatal days 0, 4, and 10. This is the first report of importin-alpha s and beta 1 expression throughout the CNS of adult mice, as well as in the developing brain, including cell type specific localization. (C) 2008 IBRO. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.neuroscience.2008.09.045

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Androgen action in the brain and spinal cord for the regulation of male sexual behaviors

Ken-ichi Matsuda, Hirotaka Sakamoto, Mitsuhiro Kawata

CURRENT OPINION IN PHARMACOLOGY   8 ( 6 )   747 - 751   2008.12

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Language：English   Publishing type：Book review, literature introduction, etc.   Publisher：ELSEVIER SCI LTD  

Circulating levels of androgens determine the sexual differentiation of the brain and spinal cord at a critical period, Although estradiol, which is converted from testosterone by aromatase action, can explain the cytological basis for the sexual dimorphism, androgen has its own regulatory mechanism to promote male-specific behavior through receptors. The central nervous system (CNS) employs a sex-specific neuronal network involving peptides and steroids for the expression of the sexual phenotype. Elucidation of the molecular mechanism along with the neuroanatomical background should guide the development of novel pharmacotherapeutics for sexual behavior dysfunction.

DOI： 10.1016/j.coph.2008.08.010

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THE SINGLE-PROLONGED STRESS PARADIGM ALTERS BOTH THE MORPHOLOGY AND STRESS RESPONSE OF MAGNOCELLULAR VASOPRESSIN NEURONS

T. Yoshii, H. Sakamoto, M. Kawasaki, H. Ozawa, Y. Ueta, T. Onaka, K. Fukui, M. Kawata

NEUROSCIENCE   156 ( 3 )   466 - 474   2008.10

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Vasopressin (AVP) plays an important role in anxiety-related and social behaviors. Single-prolonged stress (SPS) has been established as an animal acute severe stress model and has been shown to induce a lower adrenocorticotropic hormone (ACTH) response upon cortisol challenge. Here, we show results from immunoassays for AVP, ACTH, and corticosterone (CORT), and in situ hybridizations for AVP mRNA performed 7 days after SPS exposure. Immunofluorescence for AVP was also performed during the 7-day period following SPS exposure and after an additional forced swimming stress paradigm. We observed that the plasma concentrations of AVP, ACTH, and CORT were not altered by SPS; ACTH content in the pituitary and AVP mRNA expression in the supraoptic nucleus (SON) were significantly reduced by SPS. During the 7-day period following SPS, the intensity of immunoreactivity, the size of the soma, and the immunoreactive optical density of the dendrites of AVP neurons in the SON all increased. An apparent reduction in the intensity of AVP immunoreactivity was observed in the SON at 4 h after additional stress. Additional forced swimming led to a rapid increase in the dendritic AVP content only in the controls and not in the SPS-treated rats. These findings suggest that AVP is a potential biomarker for past exposure to severe stress and that alterations in AVP may affect the development of pathogenesis in stress-related disorders. (C) 2008 IBRO. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.neuroscience.2008.07.049

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Expression and intracellular distribution of the G protein-coupled receptor 30 in rat hippocampal formation

KenIchi Matsuda, Hirotaka Sakamoto, Hiroko Mori, Koji Hosokawa, Akeo Kawamura, Minoru Itose, Mayumi Nishi, Eric R. Prossnitz, Mitsuhiro Kawata

NEUROSCIENCE LETTERS   441 ( 1 )   94 - 99   2008.8

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Although the expression and distribution of nuclear estrogen receptors in the hippocampus has been described, it has been proposed that the nuclear receptors may not explain all aspects of estrogen function in the hippocampus. Recently, a G protein-coupled receptor for estrogen, GPR30, was identified as a membrane-localized estrogen receptor in several cancer cell lines. In this study, we examined the expression and intracellular distribution of GPR30 in the rat hippocampal formation. We found expression of GPR30 in pyramidal cells of CA1-3 and granule cells of the dentate gyrus at both mRNA and protein levels. Specific markers for intracellular organelles and immunoelectron microscopy revealed that GPR30 was mainly localized to the Golgi apparatus and partially in the endoplasmic reticulum of the neuron but could not detect the protein at the cell surface. Expression levels were not different among male, female in proestrus and female in estrus at the adult stage, but were higher in newborn male than newborn female. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.neulet.2008.05.108

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Steroid receptor signalling in the brain - Lessons learned from molecular imaging

M. Kawata, M. Nishi, K. Matsuda, H. Sakamoto, N. Kaku, M. Masugi-Tokita, K. Fujikawa, Y. Hirahara-Wada, K. Takanarni, H. Mori

JOURNAL OF NEUROENDOCRINOLOGY   20 ( 6 )   673 - 676   2008.6

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Language：English   Publishing type：Book review, literature introduction, etc.   Publisher：BLACKWELL PUBLISHING  

Studies with green fluorescent protein (GFP) have revealed the subcellular distribution of many steroid hormone receptors to be much more dynamic than previously thought. Fluorescence resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP) are powerful techniques with which to examine protein-protein interaction and the mobility of tagged proteins, respectively. FRET analysis revealed that steroid treatment (with corticosterone or testosterone) induces direct interaction of the glucocorticoid receptor (GR) and importin alpha in the cytoplasm and that, shortly after nuclear entry, the GR detaches from importin alpha. The mineralocorticoid receptor (MR) and androgen receptor (AR) show the same trafficking. Upon oestradiol treatment, ER alpha and ER beta in the same cell are relocalised to form a discrete pattern and are localised in the same discrete cluster (subnuclear foci). FRAP analysis showed that nuclear ER alpha and ER beta are most dynamic and mobile in the absence of the ligand, and that mobility decreases slightly after ligand treatment. Genomic as well as non-genomic actions of steroid hormones influence the cellular function of target tissues spacio-temporally.

DOI： 10.1111/j.1365-2826.2008.01727.x

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Platelet-rich plasma enhances the initial mobilization of circulation-derived cells for tendon healing

Yoshiteru Kajikawa, Toru Morihara, Hirotaka Sakamoto, Ken-Ichi Matsuda, Yasushi Oshima, Atsuhiko Yoshida, Masateru Nagae, Yuji Arai, Mitsuhiro Kawata, Toshikazu Kubo

JOURNAL OF CELLULAR PHYSIOLOGY   215 ( 3 )   837 - 845   2008.6

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Circulation-derived cells play a crucial role in the healing processes of tissue. In early phases of tendon healing processes, circulation-derived cells temporarily exist in the wounded area to initiate the healing process and decrease in number with time. We assumed that a delay of time-dependent decrease in circulation-derived cells could improve the healing of tendons. In this study, we injected platelet-rich plasma (PRP) containing various kinds of growth factors into the wounded area of the patellar tendon, and compared the effects on activation of circulation-derived cells and enhancement of tendon healing with a control group (no PRP injection). To follow the circulation-derived cells, we used a green fluorescent protein (GFP) chimeric rat expressing GFP in the circulating cells and bone marrow cells. In the PRP group, the numbers of GFP-positive cells and heat-shock protein (HSP47; collagen-specific molecular chaperone)-positive cells were significantly higher than in the control group at 3 and 7 days after injury. At the same time, the immunoreactivity for types I and III collagen was higher in the PRP group than in the control group at early phase of tendon healing. These findings suggest that locally injected PRP is useful as an activator of circulation-derived cells for enhancement of the initial tendon healing process.

DOI： 10.1002/jcp.21368

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Effects of single-prolonged stress on neurons and their afferent inputs in the amygdala

H. Cui, H. Sakamoto, S. Higashi, M. Kawata

NEUROSCIENCE   152 ( 3 )   703 - 712   2008.3

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The amygdala modulates memory consolidation with the storage of emotionally relevant information and plays a critical role in fear and anxiety. We examined changes in neuronal morphology and neurotransmitter content in the amygdala of rats exposed to a single prolonged stress (SPS) as a putative animal model for human post-traumatic stress disorder (PTSD). Rats were perfused 7 days after SPS, and intracellular injections of Lucifer Yellow were administered to neurons of the basolateral (BLA) and central amygdala (CeA) to analyze morphological changes at the cellular level. A significant increase of dendritic arborization in BLA pyramidal neurons was observed, but there was no effect on CeA neurons. Neuropeptide Y (NPY) was abundant in BLA under normal conditions. The local concentration and number of immunoreactive fibers of NPY in the BLA of SPS-exposed rats were increased compared with the control. No differences were observed in this regard in the CeA. Double immunostaining by fluorescence and electron microscopy revealed that NPY immunoreactive terminals were closely associated with calcium/calmodulin H-dependent protein kinase (CaMKII: a marker for pyramidal neurons)-positive neurons in the BLA, which were immunopositive to glucocorticoid receptor (GR) and mineralocorticoid receptor (MR). SPS had no significant effect on the expression of CaMKII and MR/GR expression in the BLA. Based on these findings, we suggest that changes in the morphology of pyramidal neurons in the BLA by SPS could be mediated through the enhancement of NPY functions, and this structural plasticity in the amygdala provides a cellular and molecular basis to understand for affective disorders. (C) 2008 IBRO. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.neuroscience.2007.12.028

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Behavior of host and graft cells in the early remodeling process of rotator cuff defects in a transgenic animal model

Yoshio Iwata, Toru Morihara, Hisakazu Tachiiri, Yoshiteru Kajikawa, Atsuhiko Yoshida, Yuji Arai, Daisaku Tokunaga, Hirotaka Sakamoto, Ken-ichi Matsuda, Masao Kurokawa, Mitsuhiro Kawata, Toshikazu Kubo

JOURNAL OF SHOULDER AND ELBOW SURGERY   17 ( 1 )   101S - 107S   2008.1

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Language：English   Publisher：MOSBY-ELSEVIER  

Autologous tissue graft is one of the treatment options for a large rotator cuff defect. To develop appropriate strategies for enhanced solid graft integration at the bone-tendon interface and tendon-tendon interface, clarifying the fate of the graft and host cells that contribute to repair and remodeling is necessary. We have developed a new grafting model using green fluorescent protein-transgenic rats and wild-type rats to simulate autologous transplantation for examining the behavior of the host and graft cells in the remodeling process after tendon grafting. We found that the host cells commenced proliferation in the graft at 1 day after grafting. The host cells infiltrated into the graft from the subacromial synovium, proximal tendon, and bone-tendon insertion. The number of graft-derived cells decreased with time. Our result clearly demonstrated that host cells, rather than graft cells, were essential for rotator cuff remodeling after tendon grafting for rotator cuff defect.

DOI： 10.1016/j.jse.2007.07.008

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Mode of action and functional significance of estrogen-inducing dendritic growth, spinogenesis, and synaptogenesis in the developing purkinje cell

Katsunori Sasahara, Hanako Shikimi, Shogo Haraguchi, Hirotaka Sakamoto, Shin-ichiro Honda, Nobuhiro Harada, Kazuyoshi Tsutsui

JOURNAL OF NEUROSCIENCE   27 ( 28 )   7408 - 7417   2007.7

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Neurosteroids are synthesized de novo from cholesterol in the brain. To understand neurosteroid action in the brain, data on the regio- and temporal-specific synthesis of neurosteroids are needed. Recently, we identified the Purkinje cell as an active neurosteroidogenic cell. In rodents, this neuron actively produces several neurosteroids including estradiol during neonatal life, when cerebellar neuronal circuit formation occurs. Estradiol may be involved in cerebellar neuronal circuit formation through promoting neuronal growth and neuronal synaptic contact, because the Purkinje cell expresses estrogen receptor-beta (ER beta). To test this hypothesis, in this study we examined the effects of estradiol on dendritic growth, spinogenesis, and synaptogenesis in the Purkinje cell using neonatal wild-type (WT) mice or cytochrome P450 aromatase knock-out (ArKO) mice. Administration of estradiol to neonatal WT or ArKO mice increased dendritic growth, spinogenesis, and synaptogenesis in the Purkinje cell. In contrast, WT mice treated with tamoxifen, an ER antagonist, or ArKO mice exhibited decreased Purkinje dendritic growth, spinogenesis, and synaptogenesis at the same neonatal period. To elucidate the mode of action of estradiol, we further examined the expression of brain-derived neurotrophic factor ( BDNF) in response to estrogen actions in the neonate. Estrogen administration to neonatal WT or ArKO mice increased the BDNF level in the cerebellum, whereas tamoxifen decreased the BDNF level in WT mice similar to ArKO mice. BDNF administration to tamoxifen-treated WT mice increased Purkinje dendritic growth. These results indicate that estradiol induces dendritic growth, spinogenesis, and synaptogenesis in the developing Purkinje cell via BDNF action during neonatal life.

DOI： 10.1523/JNEUROSCI.0710-07.2007

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GFP chimeric models exhibited a biphasic pattern of mesenchymal cell invasion in tendon healing

Yoshiteru Kajikawa, Toru Morihara, Nobuyoshi Watanabe, Hirotaka Sakamoto, Ken-Ichi Matsuda, Masashi Kobayash, Yasushi Oshima, Atsuhiko Yoshida, Mitsuhiro Kawata, Toshikazu Kubo

JOURNAL OF CELLULAR PHYSIOLOGY   210 ( 3 )   684 - 691   2007.3

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Language：English   Publisher：WILEY-LISS  

The healing of an injured musculoskeletal system requires an influx of mesenchymal cells that can differentiate into osteoblasts, fibroblasts, chondroblasts, and skeletal myoblasts. However, whether these mesenchymal cells arise from the circulation (bone marrow) or the injured tissues themselves has been controversial. To reveal the spatiotemporal characteristics of the reparative mesenchymal cells, we investigated the healing process after patellar tendon injury using two types of green fluorescent protein (GFP) chimeric rats; one expressing GFP in the circulating cells, and the other expressing it in the patellar tendon. We analyzed the behavior of GFP-positive cells after experimental tendon injury in both chimeric rats to clarify the origin of reparative cells. At 24 h after the injury, the wound contained circulation-derived cells but not tendon-derived cells. Tendon-derived cells first appeared in the wounded area at 3 days after the injury, and had significantly increased in number with time and had maintained a high level of proliferative activity until 7 days after the injury, whereas the circulation-derived cells had decreased in number and had been replaced by the tendon-derived cells. These findings suggest that circulation-derived and tendon-derived cells contribute to the healing of tendons in different periods as part of a biphasic process.

DOI： 10.1002/jcp.20876

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Intervertebral disc regeneration using platelet-rich plasma and biodegradable gelatin hydrogel microspheres

Masateru Nagae, Takumi Ikeda, Yasuo Mikami, Hitoshi Hase, Hitoshi Ozawa, Ken-Ichi Matsuda, Hirotaka Sakamot, Yasuhiko Tabata, Mitsuhiro Kawata, Toshikazu Kubo

TISSUE ENGINEERING   13 ( 1 )   147 - 158   2007.1

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Language：English   Publisher：MARY ANN LIEBERT INC  

This study evaluated the regenerative effects of platelet-rich plasma (PRP) for the degenerated intervertebral disc (IVD) in vivo. After induction of IVD degeneration in rabbits, we prepared PRP by centrifuging blood obtained from these rabbits. These PRP were injected into the nucleus pulposus (NP) of the degenerated IVDs after impregnation into gelatin hydrogel microspheres that can immobilize PRP growth factors physiochemically and release them in a sustained manner with the degradation of the microspheres. As controls, microspheres impregnated with phosphate-buffered saline (PBS) and PRP without microspheres were similarly injected. Histologically, notable progress in IVD degeneration with time courses was observed in the PBS control, PRP-only, and sham groups. In contrast, progress was remarkably suppressed over the 8-week period in the PRP group. Moreover, in immunohistochemistry, intense immunostaining for proteoglycan in the NP and inner layer of the annulus fibrosus was observed 8 weeks after administration of PRP-impregnated microspheres. Almost all microspheres were indistinct 8 weeks after the injection, and there were no apparent side effects in this study. Our results suggest that the combined administration of PRP and gelatin hydrogel microspheres into the IVD may be a promising therapeutic modality for IVD degeneration.

DOI： 10.1089/ten.2006.0042

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内分泌学的PTSDストレスモデルはバソプレッシン系にも影響する

吉井 崇喜, 坂本 浩隆, 川崎 展, 小澤 一史, 尾仲 達史, 上田 陽一, 福居 顯二, 河田 光博

日本内分泌学会雑誌   82 ( 2 )   368 - 368   2006.9

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Language：Japanese   Publisher：(一社)日本内分泌学会  

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The vasopressin dynamism is altered by PTSD model stress

Takanobu Yoshii, Hirotaka Sakamoto, Makoto Kawasaki, Hitoshi Ozawa, Yoichi Ueta, Kenji Fukui, Mitsuhiro Kawata

FRONTIERS IN NEUROENDOCRINOLOGY   27 ( 1 )   46 - 46   2006.5

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

DOI： 10.1016/j.yfrne.2006.03.094

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Distribution of gastrin releasing peptide in the rat lumbar spinal cord is sexually dimorphic and regulated by androgen

Hirotaka Sakamoto, Mitsuhiro Kawata

FRONTIERS IN NEUROENDOCRINOLOGY   27 ( 1 )   146 - 146   2006.5

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

DOI： 10.1016/j.yfrne.2006.03.254

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Intrauterine proximity to male fetuses affects the morphology of the sexually dimorphic nucleus of the preoptic area in the adult rat brain

M Pei, K Matsuda, H Sakamoto, M Kawata

EUROPEAN JOURNAL OF NEUROSCIENCE   23 ( 5 )   1234 - 1240   2006.3

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Language：English   Publisher：BLACKWELL PUBLISHING  

Previous studies on polytocous rodents have revealed that the fetal intrauterine position influences its later anatomy, physiology, reproductive performance and behavior. To investigate whether the position of a fetus in the uterus modifies the development of the brain, we examined whether the structure of the sexually dimorphic nucleus of the preoptic area (SDN-POA) of rat brains accorded to their intrauterine positions. Brain sections of adult rats gestated between two male fetuses (2M) and between two female fetuses (2F) in the uterus were analysed for their immunoreactivity to calbindin-D28k, which is a marker of the SDN-POA. The SDN-POA volume of the 2M adult males was greater than that of the 2F adult males, whereas the SDN-POA volume of the 2M and 2F adult females showed no significant difference. This result indicated that contiguous male fetuses have a masculinizing effect on the SDN-POA volume of the male. To further examine whether the increment of SDN-POA volume in adulthood was due to exposure to elevated steroid hormones during fetal life, concentrations of testosterone and 17 beta-estradiol in the brain were measured with 2M and 2F fetuses during gestation, respectively. On gestation day 21, the concentrations of testosterone and 17 beta-estradiol in the brain were significantly higher in the 2M male rats as compared with the 2F male rats. The results suggested that there was a relationship between the fetal intrauterine position, hormone transfer from adjacent fetuses and the SDN-POA volume in adult rat brains.

DOI： 10.1111/j.1460-9568.2006.04661.x

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Effects of single-prolonged stress on dendritic arborization of neurons in the central and basolateral amygdala

Honghai Cui, Hirotaka Sakamoto, Mitsuhiro Kawata

NEUROSCIENCE RESEARCH   55   S180 - S180   2006

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多血小板血漿含浸ゼラチンハイドロゲル粒子の椎間板内投与による椎間板再生効果の組織学的検討 Reviewed

長江将輝, 池田巧, 三上靖夫, 長谷斉, 小澤一史, 松田賢一, 坂本浩隆, 田畑泰彦, 久保俊一, 河田光博

第111 回日本解剖学会総会・全国学術集会（2006.3.29－31. 相模原）   2006

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PTSD model stress changes the vasopressin dynamism

Yoshii Takanobu, Hirotaka Sakamoto, Makoto Kawasaki, Hitoshi Ozawa, Yoichi Ueta, Kenji Fukui, Mitsuhiro Kawata

NEUROSCIENCE RESEARCH   55   S91 - S91   2006

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発達期の小脳プルキンエ細胞における25-Dxの発現 ― プルキンエ細胞が合成するプロゲステロンの作用機構 ―

筒井和義ら, 番目

生体の科学   56:296–302   2006

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DOI： 10.11477/mf.2425100403

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Dendritic growth, spinogenesis and synaptogenesis in response to neurosteroids in the developing Purkinje cell

Kazuyoshi Tsutsui, Hirotaka Sakamoto, Katsunori Sasahara, Hanako Shikimi, Kazuyoshi Ukena, Mitsuhiro Kawata

NEUROSCIENCE RESEARCH   55   S27 - S27   2006

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IIA-21 GFP骨髄キメラを用いた腱修復における未分化間葉系細胞の起源の解析(技術,一般演題発表,第46回日本組織細胞化学会総会・学術集会)

梶川 佳照, 坂本 浩隆, 松田 賢一, 渡邉 信佳, 大島 康史, 吉田 敦彦, 久保 俊一, 河田 光博

日本組織細胞化学会総会プログラムおよび抄録集   ( 46 )   90 - 90   2005

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Expression and localization of 25-Dx, a membrane-associated putative progesterone-binding protein, in the developing Purkinje cell

H Sakamoto, K Ukena, H Takemori, M Okamoto, M Kawata, K Tsutsui

NEUROSCIENCE   126 ( 2 )   325 - 334   2004

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Neurosteroids are synthesized de novo in the brain and the cerebellar Purkinje cell is a major site for neurosteroid formation. We have demonstrated that the rat Purkinje cell actively produces progesterone de novo from cholesterol only during neonatal life and progesterone promotes dendritic growth, spinogenesis and synaptogenesis via its nuclear receptor in this neuron. On the other hand, 25-Dx, a putative membrane progesterone receptor, has been identified in the rat liver. In this study, we therefore investigated the expression and localization of 25-Dx in the Purkinje cell to understand the mode of progesterone actions in this neuron. Reverse transcription-PCR and Western immunoblot analyses revealed the expressions of 25-Dx mRNA and 25-Dx-like protein in the rat cerebellum, which increased during neonatal life. By immunocytochemistry, the expression of 25-Dx-like protein was localized in the Purkinje cell and external granule cell layer. At the ultrastructural level, we further found that 25-Dx-like immunoreactivity was associated with membrane structures of the endoplasmic reticulum and Golgi apparatus in the Purkinje cell. These results indicate that the Purkinje cell expresses the putative membrane progesterone receptor, 25-Dx during neonatal life. Progesterone may promote dendritic growth, spinogenesis and synaptogenesis via 25-Dx as well as its nuclear receptor in the Purkinje cell in the neonate. (C) 2004 IBRO. Published by Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.neuroscience.2004.04.003

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ANALYSIS OF THE ROLE OF ESTROGEN IN THE GROWTH OF CEREBELLAR PURKINJE CELLS USING AROMATASE-DEFICIENT MICE

SHIKIMI Hanako, SAKAMOTO Hirotaka, UKENA Kazuyoshi, HARADA Nobuhiro, TSUTSUI Kazuyoshi

( 18 )   15 - 15   2003.8

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Neonatal expression of progesterone receptor isoforms in the cerebellar Purkinje cell in rats

H Sakamoto, H Shikimi, K Ukena, K Tsutsui

NEUROSCIENCE LETTERS   343 ( 3 )   163 - 166   2003.6

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The cerebellar Purkinje cell (PC) is a typical site for neurosteroid formation. Progesterone synthesized de novo in developing PCs participates in the promotion of dendritic growth, spinogenesis and synaptogenesis in this neuron and such organizing actions may contribute to the formation of the cerebellar neuronal circuit during rat neonatal life. Progesterone receptors (PR) occur as two isoforms (PR-A and PR-B) derived from a single gene. To clarify the mode of organizing actions of progesterone, therefore, we examined the expression of these PR isoforms in the rat cerebellum during development. Western immunoblot analysis revealed that both PR isoforms were expressed highly in the cerebellum during neonatal life and the expression decreased thereafter. PR-like immunoreactivity was localized primarily in PCs in the neonatal cerebellum. Thus, progesterone may act directly on PCs via PR isoforms to promote its dendritic growth, spinogenesis and synaptogenesis. (C) 2003 Elsevier Science Ireland Ltd. All rights reserved.

DOI： 10.1016/S0304-9340(03)000362-8

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エストロゲンが誘導する小脳プルキンエ細胞の発達:アロマターゼノックアウトによる解析

SHOKUMI HANAKO, SAKAMOTO HIROTAKA, UKIANA KAZUYOSHI, HARADA NOBUHIRO, TSUTSUI KAZUYOSHI

日本比較内分泌学会大会及びシンポジウムプログラム・講演要旨   28th   25   2003

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ENDCRINE DISRUPTING CHEMICALS PROMOTE PURKINJE DENDRITIC GROWTH DURING CEREBELLAR CORTICAL FORMATION(Endocrinology,Abstracts of papers presented at the 74^<th> Annual Meeting of the Zoological Society of Japan) :

Shikimi Hanako, Sakamoto Hirotaka, Mezaki Yukio, Ukena Kazuyoshi, Tsutsui Kazuyoshi

Zoological science   20 ( 12 )   1601 - 1601   2003

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Language：English   Publisher：Zoological Society of Japan  

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EXPRESSION AND LOCALIZATION OF THE PUTATIVE MEMBRANE PROGESTERONE RECEPTOR 25-DX IN THE DEVELOPING PURKINJE CELL(Endocrinology,Abstracts of papers presented at the 74^<th> Annual Meeting of the Zoological Society of Japan) :

Tsutsui Kazuyoshi, Sakamoto Hirotaka, Shikimi Hanako, Ukena Kazuyoshi, Takemori Hiroshi, Okamoto Mitsuhiro, Kawata Mitsuhiro

Zoological science   20 ( 12 )   1606 - 1606   2003

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Dendritic spine formation in response to progesterone synthesized de novo in the developing Purkinje cell in rats

Hirotaka Sakamoto, Kazuyoshi Ukena, Kazuyoshi Tsutsui

Neuroscience Letters   322 ( 2 )   111 - 115   2002.4

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The cerebellar Purkinje cell (PC) is a typical site for neurosteroid formation. We have demonstrated that this neuron possesses intranuclear receptor for progesterone and actively synthesizes progesterone de novo from cholesterol only during rat neonatal life, when the formation of the cerebellar cortex occurs dramatically. In this study, we therefore analyzed the effect of progesterone on dendritic spine formation of the PC. In vitro studies using cerebellar slice cultures from newborn rats showed that progesterone increases the density of PC dendritic spines in a dose-dependent manner. This effect was blocked by the progesterone receptor antagonist, RU486. Furthermore, trilostane, a specific inhibitor of progesterone synthesis, inhibited the increase of spine density. These results suggest that progesterone can promote dendritic spine formation, and endogenous progesterone synthesized de novo in the developing PC may induce such an effect. © 2002 Elsevier Science Ireland Ltd. All rights reserved.

DOI： 10.1016/S0304-3940(02)00077-0

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Organizing Actions of Neurosteroids Synthesized De Novo in the Cerebellar Purkinje Neuron

SAKAMOTO Hirotaka

Memoirs of the Faculty of Integrated Arts and Sciences, Hiroshima University. IV, Science reports : studies of fundamental and environmental sciences   28   141 - 144   2002

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Language：Japanese   Publisher：Hiroshima University  

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ACTIVITY AND LOCALIZATION OF 3β-HYDROXYSTEROID DEHYDROGENASE/Δ^5-Δ^4- ISOMERASE IN THE ZEBRAFISH BRAIN

SAKAMOTO Hirotaka, UKENA Kazuyoshi, TSUTSUI Kazuyoshi

16   23 - 23   2001.12

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SEX STEROIDS SYNTHESIZED DE NOVO IN THE DEVELOPING PURKINJE NEURON PROMOTE DENDRITIC GROWTH DURING CEREBELLAR CORTICAL FORMATION

MEZAKI Yukio, SAKAMOTO Hirotaka, UKENA Kazuyoshi, TSUTSUI Kazuyoshi

16   26 - 26   2001.12

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Effects of progesterone synthesized de novo in the developing Purkinje cell on its dendritic growth and synaptogenesis

坂本浩隆他

J. Neurosci.   21:6221-6232   2001

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Activity and localization of 3β-hydroxysteroid dehydrogenase/Δ5-Δ4-isomerase in the zebrafish central nervous system

坂本浩隆他

J. Comp. Neurol.   439:291-305.   2001

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Existence of galanin in lumbosacral sympathetic ganglionic neurons that project to the quail uterine oviduct

H Sakamoto, T Ubuka, C Kohchi, D Li, K Ukena, K Tsutsui

ENDOCRINOLOGY   141 ( 12 )   4402 - 4412   2000.12

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Oviposition in birds is conducted by vigorous contractions of the uterine oviduct. We recently isolated an oviposition-inducing peptide that was identified as avian galanin from mature quail oviducts. This peptide was localized in neuronal fibers terminating in muscle layers in the uterine oviduct and evoked vigorous uterine contractions through binding to receptors located in the uterus. However, no cell bodies that express avian galanin were detected in the uterus or other oviduct regions. To understand the control mechanism of avian oviposition by galanin, me identified the neurons that synthesize galanin and project to the uterus with the combination of retrograde labeling with neurobiotin and immunocytochemistry for galanin in mature Japanese quails. Retrograde labeling with neurobiotin from the uterus revealed that lumbosacral sympathetic ganglionic neurons located in the uterine side projected their axons to the uterine muscle layer. Abundant elementary granules were observed in somata of the retrogradely labeled sympathetic ganglionic neurons, suggesting that labeled neurons may function as a neurosecretory cell. Immunocytochemical analysis with the antiserum against avian galanin showed an intense immunoreaction restricted to somata of the retrograde-labeled ganglionic neurons. Preabsorbing the antiserum with avian galanin resulted in a complete absence of the immunoreaction. Competitive enzyme-linked immunosorbent assay using antigalanin serum confirmed that avian galanin existed in the sympathetic ganglionic neurons. Expression of the avian galanin messenger RNA in the neurons was further verified by Northern blot analysis. In addition, both avian galanin and its messenger RNA in the neurons were highly expressed in mature birds, unlike in immature birds.
These results suggest that lumbosacral sympathetic ganglionic neurons innervating the uterine muscle produce avian galanin in mature birds. Because this peptide acts directly on the uterus to evoke oviposition through a mechanism of the induction of vigorous uterine contraction, galaninergic innervation of the uterine oviduct may be essential for avian oviposition.

DOI： 10.1210/en.141.12.4402

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腰仙椎交感神経節ニューロンにおける産卵誘起ペプチドの発現と発達過程における変動

産賀崇由, 坂本浩隆, 河内千恵, 浮穴和義, 筒井和義

日本動物学会中国四国支部会報   ( 52 )   2000

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鳥類の腰仙椎交感神経節ニューロンにおける産卵誘起ペプチドの発現と性ステロイドによる誘導

産賀崇由, 坂本浩隆, 浮穴和義, 筒井和義

Annual Meeting of Japanese Avian Endocrinology   25th   2000

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交感神経節ニューロンにおける産卵誘起ペプチドの発現と性ステロイドによる誘導

産賀崇由, 坂本浩隆, 浮穴和義, 筒井和義

日本動物学会大会予稿集   71st   2000

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Development of the myotomal neuromuscular system in embryonic and larval angelfish, Pterophyllum scalare

H Sakamoto, M Yoshida, T Sakamoto, K Uematsu

ZOOLOGICAL SCIENCE   16 ( 5 )   775 - 784   1999.10

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To reveal the developmental sequence of the myotomal neuromuscular system in a teleost angelfish, Pterophyllum scalare, we investigated the differentiation and axonal outgrowth of the somatic spinal motoneurons as well as the differentiation of the axial muscles by means of anatomical and histochemical methods. Acetylcholinesterase histochemistry and retrograde labeling with HRP revealed two large motoneurons in each spinal hemisegment in the late embryos. To clarify the posthatching change of the motoneuron number, the number of axons in the anal-level ventral root was counted, since ChAT-immunohistochemical labeling of cholinergic spinal neurons and electron microscopic observation in the adult showed that the ventral roots around the anal level contained only somatic motor axons. We found that 15 primary motoneurons in each spinal hemisegment participated in the muscle innervation in just-hatched larvae. The motor axons rapidly increased in number beyond the adult level within three days posthatching, and then decreased to reach the adult level within a few weeks. The result suggests that competition among the motoneurons for their target muscles takes place. To reveal the temporal sequence of differentiation of the myotomal muscle fibers, in addition to electron microscopic observation of the muscle, a fluorescent mitochondrial marker dimethylaminostyrylethylpyridiniumiodine (DASPEI) was used to detect red muscle fibers. In the late embryo, immature white muscle fibers subserving the twitching movement of the animal in the egg capsule were observed. Differentiation of the red muscle was not evident until day 10. The present results show that a complete set of the axial muscle motoneurons differentiates before the differentiation of the multiple muscle fiber types in the angelfish.

DOI： 10.2108/zsj.16.775

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Naturally occurring somatic motoneuron death in a teleost angelfish, Pterophyllum scalare

H Sakamoto, M Yoshida, K Uematsu

NEUROSCIENCE LETTERS   267 ( 2 )   145 - 148   1999.5

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Language：English   Publisher：ELSEVIER SCI IRELAND LTD  

Naturally occurring somatic motoneuron death in a teleost angelfish, Pterophyllum scalare, was investigated histochemically and electron microscopically. The number of motor axons in the ventral root, which corresponds to the motoneuron number in spinal hemisegment, was rapidly increased beyond the adult value within 3 days after hatching, and then decreased to reach the adult value within a few weeks. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) histochemistry, which detects fragmented nuclear DNA characteristic to apoptotic cells, showed that the apoptotic cells are located in the motor column of the cord in the larvae at specific developmental stages. Electron microscopic observations of the spinal cells further confirmed the motoneuron apoptosis. The present data suggest that the massive death of somatic motoneurons at certain ontogenic stages which has been known to occur in higher vertebrates also takes place in fish. (C) 1999 Elsevier Science ireland Ltd. All rights reserved.

DOI： 10.1016/S0304-3940(99)00353-5

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DEVELOPMENT OF SPINAL MOTONEURONS INNERVATING THE AXIAL MUSCULATURE IN EMBRYONIC AND LARVAL ANGELFISH(Physiology)(Proceedings of the Sixty-Ninth Annual Meeting of the Zoological Society of Japan) :

Sakamoto H., Sakamoto T., Yoshida M., Uematsu K.

Zoological science   15   101 - 101   1998

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Other Link： http://id.nii.ac.jp/1141/00033857/