研究者詳細 - 木村　康二

The effects on the endocrine system under hepatotoxicity induction by phenobarbital and di(2-ethylhexyl)phthalate in intact juvenile male rats. 査読

Yamaguchi T, Maeda M, Ogata K, Abe J, Utsumi T, Kimura K

The Journal of toxicological sciences 44 ( 7 ) 459 - 469 2019年

Involvement of interferon-tau in the induction of apoptotic, pyroptotic, and autophagic cell death-related signaling pathways in the bovine uterine endometrium during early pregnancy. 査読

Toshiyuki Suzuki, Ryosuke Sakumoto, Ken-Go Hayashi, Takatoshi Ogiso, Hiroki Kunii, Takahiro Shirozu, Sung-Woo Kim, Hanako Bai, Manabu Kawahara, Koji Kimura, Masashi Takahashi

The Journal of reproduction and development 64 ( 6 ) 495 - 502 2018年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

Interferon-tau (IFNT), a type I interferon (IFN), is known as pregnancy recognition signaling molecule secreted from the ruminant conceptus during the preimplantation period. Type I IFNs, such as IFN-alpha and IFN-beta, are known to activate cell-death pathways as well as induce apoptosis. In cows, induction of apoptosis with DNA fragmentation is induced by IFNT in cultured bovine endometrial epithelial cells. However, the status of cell-death pathways in the bovine endometrium during the preimplantation period still remains unclear. In the present study, we investigated the different cell-death pathways, including apoptosis, pyroptosis, and autophagy, in uterine tissue obtained from pregnant cows and in vitro cultured endometrial epithelial cells with IFNT stimulation. The expression of CASP7, 8, and FADD (apoptosis-related genes) was significantly higher in pregnant day 18 uterine tissue in comparison to non-pregnant day 18 tissue. The expression of CASP4, 11, and NLRP3 (pyroptosis-related genes) was significantly higher in the pregnant uterus in comparison to non-pregnant uterus. In contrast, autophagy-related genes were not affected by pregnancy. We also investigated the effect of IFNT on the expression of cell-death pathway-related genes, as well as DNA fragmentation in cultured endometrial epithelial cells. Similar to its effects in pregnant uterine tissue, IFNT affected the increase of apoptosis-related (CASP8) and pyroptosis-related genes (CASP11), but did not affect autophagy-related gene expression. IFNT also increased γH2AX-positive cells, which is a marker of DNA fragmentation. These results suggest that apoptosis- and pyroptosis-related genes are induced by IFNT in the pregnant bovine endometrial epithelial cells.

DOI： 10.1262/jrd.2018-063

Propionate and butyrate induce gene expression of monocarboxylate transporter 4 and cluster of differentiation 147 in cultured rumen epithelial cells derived from preweaning dairy calves. 査読国際誌

Nakamura S, Haga S, Kimura K, Matsuyama S

Journal of animal science 96 ( 11 ) 4902 - 4911 2018年9月

子宮内膜における免疫系は暑熱ストレスにより変化する

酒井駿介, 畑生俊光, 山本ゆき, 木村康二

The Journal of Reproduction and Development 64 ( Suppl. ) j83 - j83 2018年9月

記述言語：日本語出版者・発行元：(公社)日本繁殖生物学会

【目的】分娩後ほぼすべてのウシは子宮腔への細菌侵入により子宮内膜炎を発症する。これに対し，子宮内膜細胞はIL-6およびIL-8を分泌し，主要感染部位である子宮内膜上皮層へマクロファージ（MΦ）および好中球を誘導することで細菌除去し，子宮内膜炎を治癒する。しかし，夏季に子宮内膜炎を発症した場合，症状が長期化すると報告されており，夏季不妊の一因として考えられる。本研究では，夏季の子宮内膜炎の長期化はHeat Stress（HS）による子宮内膜の免疫機能低下によると仮定し，冬季および夏季の子宮内膜組織におけるMΦおよび好中球の局在解析と，培養子宮内膜細胞を用いた，リポ多糖（LPS）刺激によるIL-6およびIL-8産生へのHSの影響を検討した。【方法】冬季および夏季に採取した子宮内膜においてMΦおよび好中球の局在を免疫組織化学およびギムザ染色により検討した。また，培養子宮内膜上皮および間質細胞にLPS添加後，38.5°Cまたは40.5°Cで培養し，IL-6およびIL-8遺伝子発現および上清中の濃度を定量的RT-PCR法もしくはEIAによりそれぞれ測定した。【結果】MΦは冬季の子宮内膜において上皮層付近に局在するが，夏季には子宮内膜全域に散在した。一方，好中球の局在は冬季と夏季で変化がなかった。また，38.5°Cで培養した上皮細胞においてIL-6およびIL-8遺伝子発現および分泌量はLPSにより有意に増加した。このIL-6増加はHSにより有意に抑制されたが，IL-8増加にHSは影響しなかった。一方，38.5°Cで培養した間質細胞においてIL-6およびIL-8遺伝子発現および産生量はLPS添加により有意に増加し，この増加はHSによって増強された。以上より，HSは子宮内膜細胞による好中球の誘引に影響しない一方，上皮細胞のIL-6産生を抑制し，間質細胞のIL-6産生を刺激することで上皮層へのMΦ誘導を抑制する可能性が考えられた。これにより夏季の子宮内膜炎症状を長期化させ，夏季不妊の一因となることが示唆された。

DOI： 10.14882/jrds.111.0_or2-13

CiNii Article

Heat stress affects prostaglandin synthesis in bovine endometrial cells. 査読

Shunsuke Sakai, Natsumi Hagihara, Mariko Kuse, Koji Kimura, Kiyoshi Okuda

The Journal of reproduction and development 64 ( 4 ) 311 - 317 2018年8月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Heat stress (HS) negatively affects reproduction in cattle; however, its effect on endocrine function in bovine endometrial cells remains unclear. In this study, we examined the effects of HS on the production of prostaglandin (PG) E2 and PGF2α in the cultured bovine endometrial epithelial and stromal cells separately. To evaluate the effect of HS on endocrine function, the cells were cultured at 38.5°C (control) or 40.5°C (HS). After treatment, PGE2 and PGF2α levels were measured via enzyme immunoassay (EIA) and mRNA expressions of enzymes involved in PG synthesis were examined via quantitative reverse transcription polymerase chain reaction (RT-PCR). HS did not influence the production of PGE2 or PGF2α in the epithelial cells; however, HS significantly enhanced the production of both PGE2 and PGF2α in the stromal cells (P < 0.05). In addition, HS significantly increased phospholipase A2 (PLA2), cyclooxygenase 2 (COX2), prostaglandin F synthase (PGFS), prostaglandin E synthase (PGES), and carbonyl reductase 1 (CBR1) mRNA expression in the stromal cells (P < 0.05). The overall results suggest that HS induces mRNA expression of enzymes involved in PG synthesis, resulting in the upregulation of PGE2 and PGF2α production in the stromal cells, but not in the epithelial cells. The HS-induced increase of PGE2 and PGF2α secretion in bovine endometrial stromal cells may disrupt the normal estrous cycle and cause infertility in cows during summer.

DOI： 10.1262/jrd.2018-051

ウシ黄体における骨形成蛋白質結合蛋白質の役割

西村亮, 長谷川啓喜, 山下真路, 伊藤典彦, 岡本芳晴, 木村康二, 奥田潔

日本獣医学会学術集会講演要旨集 161回 404 - 404 2018年8月

Activation of lysosomal cathepsins in pregnant bovine leukocytes. 査読国際誌

Md Abdus Shabur Talukder, Ahmed Zaky Balboula, Takahiro Shirozu, Sung Woo Kim, Hiroki Kunii, Toshiyuki Suzuki, Tsukino Ito, Koji Kimura, Masashi Takahashi

Reproduction (Cambridge, England) 155 ( 6 ) 515 - 528 2018年6月

記述言語：英語掲載種別：研究論文（学術雑誌）

In ruminants, interferon-tau (IFNT)-mediated expression of interferon-stimulated genes in peripheral blood leukocytes (PBLs) can indicate pregnancy. Recently, type 1 IFN-mediated activation of lysosomes and lysosomal cathepsins (CTSs) was observed in immune cells. This study investigated the status of lysosomal CTSs and lysosomes in PBLs collected from pregnant (P) and non-pregnant (NP) dairy cows, and conducted in vitro IFNT stimulation of NP blood leukocytes. Blood samples were collected 0, 7, 14 and 18 days post-artificial insemination, and the peripheral blood mononuclear cells (PBMCs) and polymorphonuclear granulocytes (PMNs) separated. The fluorescent activity of CTSB and CTSK in PMNs significantly increased with the progress of pregnancy, especially on day 18. In vitro supplementation of IFNT significantly increased the activities of CTSB and CTSK in NP PBMCs and PMNs. CTSB expression was significantly higher in PBMCs and PMNs collected from P day-18 cows than from NP cows, whereas there was no difference in CTSK expression. IFNT increased CTSB expression but did not affect CTSK expression. Immunodetection showed an increase of CTSB in P day-18 PBMCs and PMNs. In vitro stimulation of IFNT increased CTSB in NP PBMCs and PMNs. Lysosomal acidification showed a significant increase in P day-18 PBMCs and PMNs. IFNT also stimulated lysosomal acidification. Expressions of lysosome-associated membrane protein (LAMP) 1 and LAMP2 were significantly higher in P day-18 PBMCs and PMNs. The results suggest that pregnancy-specific activation of lysosomal functions by CTS activation in blood leukocytes is highly associated with IFNT during maternal and fetal recognition of pregnancy.

DOI： 10.1530/REP-18-0078

Down-regulation of transcription factor OVOL2 contributes to epithelial-mesenchymal transition in a noninvasive type of trophoblast implantation to the maternal endometrium. 査読国際誌

Rulan Bai, Kazuya Kusama, Keigo Nakamura, Toshihiro Sakurai, Koji Kimura, Atsushi Ideta, Yoshito Aoyagi, Kazuhiko Imakawa

FASEB journal : official publication of the Federation of American Societies for Experimental Biology 32 ( 6 ) 3371 - 3384 2018年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：FASEB

Embryo implantation into the uterine endometrium is required for pregnancy establishment in most mammals. By using global expression analysis, we investigated the molecules that are related to epithelial-mesenchymal transition (EMT) in noninvasive bovine trophoblasts and found that the transcription factor, ovo-like zinc finger 2 ( OVOL2), which is essential for mesenchymal-epithelial transition in various cancers, was down-regulated after trophoblast attachment to the endometrial epithelium in utero. In cultured bovine trophoblast cells, OVOL2 down-regulation occurred only when cells were allowed to attach to bovine endometrial epithelial cells via the TEAD3/YAP signaling pathway. This resulted in the up-regulation of the EMT-associated transcription factors, ZEB1 and SNAI2, and the mesenchymal cell markers, N-cadherin ( CDH2) and vimentin ( VIM), whereas epithelial cell marker, E-cadherin ( CDH1), was down-regulated. In contrast, OVOL2 overexpression in bovine trophoblast cells exhibited a decrease in ZEB1 transcripts and an increase in E-cadherin. These observations revealed that ovo-like protein (OVOL)2 down-regulation occurred concurrently with conceptus implantation into the uterine endometrium via the YAP/TEAD3 signaling pathway, and suggest that the down-regulation of OVOL2 expression contributes to the up-regulation of EMT-related transcription factor expression, which enables EMT progression in the noninvasive bovine trophectoderm postimplantation.-Bai, R., Kusama, K., Nakamura, K., Sakurai, T., Kimura, K., Ideta, A., Aoyagi, Y., Imakawa, K. Down-regulation of transcription factor OVOL2 contributes to epithelial-mesenchymal transition in a noninvasive type of trophoblast implantation to the maternal endometrium.

DOI： 10.1096/fj.201701131RR

Scopus

Exchange protein directly activated by cAMP (EPAC) promotes transcriptional activation of the decidual prolactin gene via CCAAT/enhancer-binding protein in human endometrial stromal cells. 査読

Bai R, Kusama K, Nakamura K, Sakurai T, Kimura K, Ideta A, Aoyagi Y, Imakawa K

FASEB JOURNAL 32 ( 6 ) 3371 - 3384 2018年6月

Publisher Correction: Correction of a Disease Mutation using CRISPR/Cas9-assisted Genome Editing in Japanese Black Cattle. 査読

Ikeda M, Matsuyama S, Akagi S, Ohkoshi K, Nakamura S, Minabe S, Kimura K, Hosoe M

Scientific reports 8 ( 1 ) 1470 2018年1月

ウシ子宮内膜におけるミトコンドリア機能の亢進は受胎率を飛躍的に向上させる

松山秀一, 中村翔, 美辺詩織, 岩田尚孝, 木村康二

日本胚移植学雑誌 40 ( 1 ) 24 - 24 2018年1月

Acute stimulation of a smooth muscle constrictor by oestradiol-17β via GPER1 in bovine oviducts 査読

Nishie T, Kobayashi Y, Kimura K, Okuda K

Reproduction in Domestic Animals 53 ( 2 ) 326 - 332 2018年

Correction of a Disease Mutation using CRISPR/Cas9-assisted Genome Editing in Japanese Black Cattle 査読

Mitsumi Ikeda, Shuichi Matsuyama, Satoshi Akagi, Katsuhiro Ohkoshi, Sho Nakamura, Shiori Minabe, Koji Kimura, Misa Hosoe

SCIENTIFIC REPORTS 7 ( 1 ) 17827 2017年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：NATURE PUBLISHING GROUP

Isoleucyl-tRNA synthetase (IARS) syndrome is a recessive disease of Japanese Black cattle caused by a single nucleotide substitution. To repair the mutated IARS gene, we designed clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) to create a double-strand break near the mutation site. CRISPR/Cas9 and donor DNA that contained a synonymous codon for the correct amino acid and an Aequorea coerulescens Green Fluorescent Protein (AcGFP) cassette with a piggyBac transposase recognition site at both ends were introduced into bovine fetal fibroblast (BFF) cells isolated from a homozygous mutant calf. Recombinant cells were enriched on the basis of expression of AcGFP, and two cell lines that contained the repaired allele were subcloned. We generated somatic cell nuclear transfer (SCNT) embryos from the repaired cells and transferred 22 blastocysts to recipient cows. In total, five viable fetuses were retrieved at Days 34 and 36. PiggyBac transposase mRNA was introduced into BFF cells isolated from cloned foetuses and AcGFP-negative cells were used for second round of cloning. We transferred nine SCNT embryos to recipient cows and retrieved two fetuses at Day 34. Fetal genomic DNA analysis showed correct repair of the IARS mutation without any additional DNA footprint.

DOI： 10.1038/s41598-017-17968-w

Hypoxia increases glucose transporter 1 expression in bovine corpus luteum at the early luteal stage 査読

Ryo Nishimura, Hiroki Hasegawa, Masamichi Yamashita, Norihiko Ito, Yoshiharu Okamoto, Takashi Takeuchi, Tomoaki Kubo, Kosuke Iga, Koji Kimura, Mitsugu Hishinuma, Kiyoshi Okuda

JOURNAL OF VETERINARY MEDICAL SCIENCE 79 ( 11 ) 1878 - 1883 2017年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：JAPAN SOC VET SCI

A major role of the corpus luteum (CL) is to produce progesterone (P4). The CL has immature vasculature shortly after ovulation, suggesting it exists under hypoxic conditions. Hypoxia-inducible factor-1 (HIF1) induces the expression of glucose transporter 1 (GLUT1). To clarify the physiological roles of GLUT1 in bovine CL, we examined GLUT1 mRNA expression in the CL under hypoxic conditions by quantitative RT-PCR. We also measured the effects of glucose (0-25 mM) and GLUT1 inhibitors (cytochalasin B, STF-31) on P4 production in bovine luteal cells. GLUT1 mRNA expression in bovine CL was higher at the early luteal stage compared to the other later stages. Hypoxia (3% O-2) increased GLUT1 mRNA expression in early luteal cells, but not in mid luteal cells. Glucose (0-25 mM) increased P4 production in early luteal cells, but not in mid luteal cells. Both GLUT1 inhibitors decreased P4 production in early and mid luteal cells. Overall, the results suggest that GLUT1 (possibly induced by hypoxic conditions in the early CL) plays a role in the establishment and development of bovine CL, especially in supporting luteal P4 synthesis at the early luteal stage.

DOI： 10.1292/jvms.17-0284

Interferon Tau Regulates Cytokine Production and Cellular Function in Human Trophoblast Cell Line 査読

Nao Tanikawa, Kotomi Seno, Ryouka Kawahara-Miki, Koji Kimura, Shuichi Matsuyama, Hisataka Iwata, Takehito Kuwayama, Koumei Shirasuna

JOURNAL OF INTERFERON AND CYTOKINE RESEARCH 37 ( 10 ) 456 - 466 2017年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：MARY ANN LIEBERT, INC

Type I interferons (IFN), including IFN-beta (IFNB), activate multiple STAT signaling to drive various biological responses. Another type I IFN, IFN-tau (IFNT), secreted by ruminant embryonic trophoblast cells, has multiple functions with low cytotoxicity. Here, we examined the effects of IFNT on human trophoblast cell functions. First, we performed next-generation sequencing and demonstrated that IFNT-dependent changes in the human Sw.71 trophoblast cell line are partly mediated by proinflammatory as well as IFN signaling. Next, we validated candidate genes, and data confirmed that IFNT stimulated interleukin-6 (IL-6) and IL-8 mRNA expression and secretion. However, human IFNB did not affect IL-6 and IL-8 mRNA expression and secretion. IFNT-induced cytokine secretion was dependent on STAT3 signaling, but not STAT1 signaling. In addition, treatment with IFNT, IL-6, or IL-8 increased cell proliferation, and IFNT also stimulated cell migration in human trophoblast cells. Although IFNT did not affect superoxide dismutase (SOD) 1 mRNA expression, it clearly increased mitochondrial SOD2 mRNA expression, resulting in the acceleration of SOD activity. We demonstrated that in addition to IFN signaling, IFNT also regulated inflammation-related signaling as well as cell proliferation, migration, and redox signaling in human trophoblast cells.

DOI： 10.1089/jir.2017.0057

Production of calves by the transfer of cryopreserved bovine elongating conceptuses and possible application for preimplantation genomic selection 査読

Takashi Fujii, Hiroki Hirayama, Akira Naito, Masashi Kashima, Hitomi Sakai, Shigeo Fukuda, Hitomi Yoshino, Satoru Moriyasu, Soichi Kageyama, Yoshikazu Sugimoto, Shuichi Matsuyama, Hiroyuki Hayakawa, Koji Kimura

JOURNAL OF REPRODUCTION AND DEVELOPMENT 63 ( 5 ) 497 - 504 2017年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SOCIETY REPRODUCTION & DEVELOPMENT-SRD

Preimplantation genomic selection based on single nucleotide polymorphism (SNP) genotypes is expected to accelerate genetic improvement in cattle. However, genome-wide genotyping at the early embryonic stage has several limitations, such as the technical difficulty of embryonic biopsy and low accuracy of genotyping resulting from a limited number of biopsied cells. After hatching from the zona pellucida, the morphology of the bovine embryo changes from spherical to filamentous, in a process known as elongation. The bovine nonsurgical elongating conceptus transfer technique was recently developed and applied for sexing without requiring specialized skills for biopsy. In order to develop a bovine preimplantation genomic selection system combined with the elongating conceptus transfer technique, we examined the accuracy of genotyping by SNP chip analysis using the DNA from elongating conceptuses (Experiment 1) and optimal cryopreservation methods for elongating conceptuses (Experiment 2). In Experiment 1, the call rates of SNP chip analysis following whole genome amplification in biopsied cells from two elongating conceptuses were 95.14% and 99.32%, which were sufficient for estimating genomic breeding value. In Experiment 2, the rates of dead cells in elongating conceptuses cryopreserved by slow freezing were comparable to those in fresh elongating conceptuses. In addition, we obtained healthy calves by the transfer of elongating conceptuses cryopreserved by slow freezing. Our findings indicate that the elongating conceptus transfer technology enables preimplantation genomic selection in cattle based on SNP chip analysis. Further studies on the optimization of cryopreservation methods for elongating conceptuses are required for practical application of the selection system.

DOI： 10.1262/jrd.2017-025

Age-associated mRNA expression changes in bovine endometrial cells in vitro. 査読国際誌

Nao Tanikawa, Ayaka Ohtsu, Ryouka Kawahara-Miki, Koji Kimura, Shuichi Matsuyama, Hisataka Iwata, Takehito Kuwayama, Koumei Shirasuna

Reproductive biology and endocrinology : RB&E 15 ( 1 ) 63 - 63 2017年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：BIOMED CENTRAL LTD

BACKGROUND: Endometrial cells secrete various cytokines and the dysfunction of endometrial cells may directly lead to infertility. Interferon tau (IFNT) secreted by trophoblast cells, a well-known pregnancy recognition signal in ruminants, acts on the uterus to prepare for pregnancy. Aging causes cellular and organ dysfunction, and advanced maternal age is associated with reduced fertility. However, few studies have investigated age-dependent changes in the uterus. METHODS: Using next generation sequencing and real-time PCR, we examined mRNA expression in bovine endometrial cells in vitro obtained from young (mean 45.2 months) and aged (mean 173.5 months) animals and the effects of IFNT depending on the age. RESULTS: We showed that inflammation-related (predicted molecules are IL1A, C1Qs, DDX58, NFKB, and CCL5) and interferon-signaling (predicted molecules are IRFs, IFITs, STATs, and IFNs) pathways were activated in endometrial cells obtained from aged compared to young cows. Also, the activation of "DNA damage checkpoint regulation" and the inhibition of "mitotic mechanisms" in endometrial cells obtained from aged cows were evident. Moreover, we showed lower cell viability levels in endometrial cells obtained from aged compared to young cows. Although treatment with IFNT upregulated various types of interferon stimulated genes both in endometrial cells obtained from young and aged cows, the rate of increase by IFNT stimulus was obviously lower in endometrial cells obtained from aged compared to young cows. CONCLUSIONS: Endometrial cells obtained from aged cows exhibited higher levels of inflammatory- and IFN-signaling, and dysfunction of cell division compared with young cows. In addition, a high basal level of IFN-related genes in endometrial cells of aged cows is suggested a concept of "inflammaging".

DOI： 10.1186/s12958-017-0284-z

Adrenomedullin regulates the speed of oviductal fluid flow in cattle 査読

Yuka Yoshimoto, Takumi Nishie, Sayaka Ito, Yoshihiko Kobayashi, Yuki Yamamoto, Kiyoshi Okuda, Koji Kimura

MOLECULAR REPRODUCTION AND DEVELOPMENT 84 ( 8 ) 712 - 718 2017年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：WILEY

Unidirectional flow of oviductal fluid from the ovarian to uterine side of the ampulla plays a significant role in successful pregnancy, and is produced by ciliary beating. Various systems regulate ciliary beating, such as paracrine, autocrine, and endocrine. We hypothesized that Adrenomedullin (ADM)-a peptide hormone that acts via its receptors, which are complexes of Calcitonin receptor-like receptor (CRLR) and Receptor activity-modifying protein (RAMP) 2 or 3-promotes oviductal fluid flow in the ampulla of bovine oviducts. First, we examined the expression of ADM, CRLR, RAMP2, and RAMP3 mRNAs in isolated epithelial cells throughout the estrous cycle, and the localization of ADM receptor protein constituents in the ampulla. RAMP2 expression was significantly higher in the follicular phase. Furthermore, RAMP2 protein was detected only in ciliated cells, whereas CRLR and RAMP3 were detected in all epithelial cells. The effects of ADM and an ADM antagonist on fluid-flow speed were examined using microbeads in ampullary tissue. ADM antagonist decreased bead transport speed, and this decrease was reversed by ADM. In addition, ADM recovered the bead transport speed that decreased in the absence of calcium. Overall, our results suggest that ADM contributes to the regulation of oviductal fluid flow in ampulla.

DOI： 10.1002/mrd.22852

Estrous cycle stage-dependent manner of type I interferon-stimulated genes induction in the bovine endometrium. 査読

Takahiro Shirozu, Hiroki Iwano, Takatoshi Ogiso, Toshiyuki Suzuki, Ahmed Z Balboula, Hanako Bai, Manabu Kawahara, Koji Kimura, Hitomi Takahashi, Bai Rulan, Sung-Woo Kim, Yojiro Yanagawa, Masashi Nagano, Kazuhiko Imakawa, Masashi Takahashi

The Journal of reproduction and development 63 ( 3 ) 211 - 220 2017年6月

記述言語：英語掲載種別：研究論文（学術雑誌）

Interferon tau (IFN-τ) is a ruminant-specific type I IFN secreted by a conceptus before its attachment to the uterus. IFN-τ induces the expression of IFN-stimulated genes (ISGs) via the type I IFN receptor (IFNAR), which is composed of IFNAR1 and IFNAR2 subunits in the endometrium. However, expression patterns of IFNARs during the estrous cycle have not been reported. We hypothesized that the response to a type I IFN changes along with IFNARs and the IFN-regulatory factors (IRFs) driving transcription of IFN signal-related genes and modulating a type I IFN signal during the estrous cycle. We investigated the estrous cycle stage-dependent type I IFN induction of ISGs and expression patterns of IFN signal-related genes in bovine endometrial tissues. Endometrial tissue pieces collected from bovine uteri at each estrous stage (early, mid, and late) were cultured with or without recombinant bovine IFN-α or concentrated pregnant uterine flushing (PUF) on day 18 after confirming the presence of a conceptus. IFN-α and PUF each significantly increased the expression of ISGs in endometrial tissues. The induction levels of the typical ISGs (MX1-a and ISG15) were significantly higher at the mid stage and correlated with high expression of IRFs at the mid stage. The immunostaining of IFNARs showed strong fluorescence intensities in luminal and glandular epithelia at the early and mid stages. Collectively, these results suggest that the endometrium exhibits estrous cycle stage-dependent responsiveness to type I IFN that may be associated with the expression of IFNARs and IRFs for pregnancy recognition.

DOI： 10.1262/jrd.2016-176