Faculty Profiles - MATSUYAMA Shuichi

Papers - MATSUYAMA Shuichi

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Effects of intrauterine extracellular vesicle microRNAs on embryonic gene expression in low-fertility cows. International journal

Rei Ichikawa, Koji Kimura, Sho Nakamura, Satoshi Ohkura, Shuichi Matsuyama

FASEB journal : official publication of the Federation of American Societies for Experimental Biology Vol. 38 ( 20 ) page： e70116 2024.10

Language：English Publishing type：Research paper (scientific journal)

Embryo survival and pre-implantation development depend on uterine luminal fluid, which is believed to play a role in early embryonic death and infertility in cows. Extracellular vesicles (EVs) in the uterine luminal fluid contain microRNAs (miRNAs), crucial mediators of intercellular communication. miRNAs regulate conceptus-maternal interactions and participate in embryonic development by suppressing gene expression. Therefore, we hypothesized that miRNAs in the intrauterine EVs of low-fertility cows would hinder embryonic survival and development. EVs were collected from the bovine uterine luminal fluid of both normal- and low-fertility cows 7 days post-estrus. Small RNA-sequencing analysis of miRNAs isolated from these EVs identified eight miRNAs that were highly expressed in normal-fertility cows (normal-fertility miRNAs) and eight with elevated expression in low-fertility cows (low-fertility miRNAs). These two sets of miRNAs were transfected into hatched blastocysts via lipofection. RNA-seq following lipofection with low-fertility miRNAs identified 424 differentially expressed genes (DEGs) relative to the control; in contrast, following lipofection with normal-fertility miRNAs, seven DEGs were identified. Pathway analysis of the DEGs identified following lipofection with low-fertility miRNAs revealed substantial enrichment of mitogen-activated protein kinase (MAPK) signaling. Expression of activator protein 1 (AP1) and interferon-tau (IFNT) mRNA was significantly lower in the low-fertility miRNA transfection group than in the control. IFNT is essential for maternal pregnancy recognition. Therefore, miRNAs in intrauterine EVs from low-fertility cows at 7 days post-estrus may inhibit embryo development and suppress IFNT expression by altering MAPK signaling.

DOI： 10.1096/fj.202401728R

Deterioration of mitochondrial biogenesis and degradation in the endometrium is a cause of subfertility in cows. Reviewed International journal

Shuichi Matsuyama, Sho Nakamura, Shiori Minabe, Miki Sakatani, Naoki Takenouchi, Takuya Sasaki, Yuki Inoue, Hisataka Iwata, Koji Kimura

Molecular reproduction and development 2023.1

Authorship：Last author,　Corresponding author Language：English Publishing type：Research paper (scientific journal)

To investigate possible causes of reproductive failure, we conducted global endometrial gene expression analyses in fertile and subfertile cows. Ingenuity pathway analysis showed that RICTOR and SIRT3 are significant upstream regulators for highly expressed genes in fertile cows, and are predicted to be activated upstream regulators of normal mitochondrial respiration. Canonical pathway analysis revealed that these highly expressed genes are involved in the activation of mitochondrial oxidative phosphorylation. Therefore, in subfertile cows, the inactivation of RICTOR and SIRT3 may correlate with decreased capacity of mitochondrial respiration. Furthermore, the expression levels of most mitochondrial DNA genes and nuclear genes encoding mitochondrial proteins were higher in subfertile cows. The mitochondrial DNA copy number was significantly higher in the endometrium of subfertile cows, whereas the ATP content did not differ between fertile and subfertile cows. Quantitative reverse transcription-PCR analysis demonstrated that the expression of PGC1a, TFAM, MFN1, FIS1, and BCL2L13 were significantly lower in subfertile cows. In addition, transmission electron microscopy images showed mitochondrial swelling in the endometrial cells of the subfertile cow. These results suggest that poor-quality mitochondria accumulate in the endometrium owing to a reduced capacity for mitochondrial biogenesis, fusion, fission, and degradation in subfertile cows, and may contribute to infertility.

DOI： 10.1002/mrd.23670

Impacts of trehalose supplementation on ruminal microbiota and productivity of Japanese Black heifers under heat-stressed conditions.

Morita Y, Mukaiyama A, Inoue S, Mukai K, Matsuyama S, Ohkura S

Animal bioscience 2025.1

Morphological observation and detail molecular characterization of Fischoederius elongatus (Digenea: Gastrothylacidae) from the rumen of domestic cattle in Cambodia. Invited Reviewed International coauthorship

Chinda, W., Bengthay, T., Suriyasathaporn, W., Morita, Y., Vutha, P., Ohkura, S., Matsuyama, S., Nakamura, S., Hayashi, K.

Journal of Parasitology 2025

Carnosine supplementation in cryopreservation solution improved frozen-thawed bovine embryo viability

ISHII Toshimichi, MORI-KOBAYASHI Kento, NAKAMURA Sho, OHKURA Satoshi, MATSUYAMA Shuichi

Journal of Reproduction and Development Vol. 70 ( 5 ) page： 279 - 285 2024.10

Language：English Publisher：The Society for Reproduction and Development

<p>Cryopreservation adversely affects embryo quality and viability <i>in vitro</i>.<i></i>We investigated the effects of cryopreservation solutions supplemented with the antioxidant carnosine on frozen-thawed bovine embryo viability. Bovine blastocysts were produced <i>in vitro</i> and cryopreserved using slow freezing. The rates of re-expanded and hatched blastocysts in the 50 μg/ml carnosine-supplemented group at 4, 24, and 48 h after thawing were higher than those in the control (P<i></i>< 0.05) group. In frozen-thawed embryos, cryopreservation solution supplemented with carnosine (50 μg/ml) significantly reduced reactive oxygen species (ROS) production<i></i>(P < 0.05), decreased TUNEL-positive apoptotic cells (P<i></i>< 0.05), and increased the mRNA expression of<i> BCL2 </i>(P<i></i>< 0.05), an apoptosis suppressor gene. The expression of translocase of outer mitochondrial membrane 20 (TOMM20), which is involved in protein mitochondrial transport, in the carnosine (50 μg/ml)-treated embryos was significantly higher than that in the control group (P < 0.05). ATP production in frozen-thawed embryos in the 50 μg/ml carnosine-supplemented group was significantly higher than that in the control group (P<i></i>< 0.05), however no significant difference in the total number of cells per embryo among the groups was observed. These results suggest that supplementing the cryopreservation solution with carnosine can improve the viability of frozen-thawed bovine embryos by reducing oxidative damage.</p>

DOI： 10.1262/jrd.2023-071

Capturing temperature changes on the ocular surface along with estrus and ovulation using infrared thermography in Japanese Black cows

Riho OZAKI, Seiji INOUE, Yuki YOROZUI, Rei ICHIKAWA, Naoki YAMADA, Seiya HIGASHI, Shuichi MATSUYAMA, Hiroko TSUKAMURA, Satoshi OHKURA, Yoshihisa UENOYAMA, Yasuhiro MORITA

Journal of Reproduction and Development Vol. 70 ( 1 ) page： 49 - 54 2024.1

Language：English Publishing type：Research paper (scientific journal) Publisher：Japanese Society of Animal Reproduction

<p>Pre-ovulatory follicles are cooler than the neighboring reproductive organs in cows. Thus, measuring the temperature of reproductive organs could be a useful method for predicting estrus and ovulation in cows, and the establishment of a non-invasive technique is required. In this study, we used infrared thermography (IRT) to measure ocular surface temperature as a potential surrogate for reproductive organ temperature. Five Japanese Black cows with synchronized estrus were subjected to temperature measurements in five regions of the ocular surface, including the nasal conjunctiva, nasal limbus, center cornea, temporal limbus, and temporal conjunctiva, twice a day (0800 h and 1600 h) during the experimental period. The temperatures in the five regions significantly declined in cows from estrus to ovulation. To the best of our knowledge, this study is the first to use IRT to show a temperature decrease in the ocular surface along with estrus to ovulation in Japanese Black cows.</p>

DOI： 10.1262/jrd.2022-116

Effect of a neurokinin 3 receptor-selective agonist administration on the embryos recovered from superovulated cows. International journal

Shuichi Matsuyama, Sho Nakamura, Shiori Minabe, Satoshi Ohkura, Koji Kimura

Biology of reproduction Vol. 108 ( 6 ) page： 936 - 944 2023.6

Language：English Publishing type：Research paper (scientific journal)

Superovulation (SOV) treatment of cows results in unovulated follicles and inconsistent quality of the recovered embryos. It has been demonstrated that luteinizing hormone (LH) secretion is suppressed during SOV treatment of cows, which may cause insufficient follicle development and variation in the development of recovered embryos and unovulated follicles. Pulsatile gonadotropin-releasing hormone/LH secretion is controlled by the activity of kisspeptin, neurokinin B, and dynorphin (KNDy) neurons in the arcuate nucleus in many mammals. Since neurokinin B promotes the activity of KNDy neurons, we hypothesized that senktide, a neurokinin B receptor agonist, has potential as a therapeutic drug to improve the ovulation rate and quality of recovered embryos in SOV-treated cows via stimulation of LH secretion. Senktide was administered intravenously (30 or 300 nmol/min) for 2 h, beginning from 72 h after the start of SOV treatment. LH secretion was examined before and after administration, and embryos were collected 7 d after estrus. Senktide administration increased LH secretion in SOV-treated cows. The ratios of code 1, code 1 and 2, and blastocyst stage embryos to recovered embryos were increased by senktide (300 nmol/min) administration. Moreover, the mRNA levels of MTCO1, COX7C, and MTATP6 were upregulated in recovered embryos of senktide (300 nmol/min)-administered animals. These results indicate that the administration of senktide to SOV-treated cows enhances LH secretion and upregulates the expression of genes involved in mitochondrial metabolism in embryos, thereby improving embryo development and embryo quality.

DOI： 10.1093/biolre/ioad039

A Survey of Genome-Wide Genetic Characterizations of Crossbred Dairy Cattle in Local Farms in Cambodia Reviewed

Mam Somony, Tep Bengthay, Rin Soriya, Uenoyama Yoshihisa, Matsuyama Shuichi, Ohkura Satoshi, Murase Tetsuma, Nunome Mitsuo, Morita Yasuhiro

ANIMALS Vol. 12 ( 16 ) 2022.8

Language：Japanese Publisher：Animals

To improve the dairy sector in Cambodia in the future, we aimed to reveal the genetic variation and the milk production in Cambodian crossbred dairy cattle. We calculated the percent (%) milk fat content and the average milk yield per cow (L/day) for two farms (Farm R and M) based on the farmers’ records and interviews. The crossbred cows originated from Cambodian local farmers and Thailand breeders in Farm R, whereas the crossbred cows originated in Thailand breeders in Farm M. Then, we performed genetic characterization for 75 individuals from the two farms and an individual Japanese pure Holstein-Friesian cow based on 133,705 single nucleotide polymorphisms (SNPs) obtained by the GRAS-Di method. The milk fat contents in the bulk milk in the dry season and the average milk yield per cow on Farm R were 3.77 ± 0.98% and 7.81 ± 2.66 L/day, respectively, and were higher than those on Farm M (3.35 ± 0.54% and 6.5–7.5 L/day). Cattle originating in Cambodia in Farm R possessed a unique genetic character different from cattle from Thailand in Farm M. The present study suggests that the differences in milk fat content between the two farms might be explained by the genetic differences in crossbred cows.

DOI： 10.3390/ani12162072

Gene-expression profile and postpartum transition of bovine endometrial side population cells† Reviewed

Ryoki Tatebayashi, Sho Nakamura, Shiori Minabe, Tadashi Furusawa, Ryoya Abe, Miki Kajisa, Yasuhiro Morita, Satoshi Ohkura, Koji Kimura, Shuichi Matsuyama

Biology of Reproduction Vol. 104 ( 4 ) page： 850 - 860 2021.4

Authorship：Last author,　Corresponding author Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Oxford University Press (OUP)

<title>Abstract</title>
The mechanism of bovine endometrial regeneration after parturition remains unclear. Here, we hypothesized that bovine endometrial stem/progenitor cells participate in the postpartum regeneration of the endometrium. Flow cytometry analysis identified the presence of side population (SP) cells among endometrial stromal cells. Endometrial SP cells were shown to differentiate into osteoblasts and adipocytes. RNA-seq data showed that the gene expression pattern was different between bovine endometrial SP cells and main population cells. Gene Set Enrichment Analysis identified the enrichment of stemness genes in SP cells. Significantly (false discovery rate &lt; 0.01) upregulated genes in SP cells contained several stem cell marker genes. Gene Ontology (GO) analysis of the upregulated genes in SP cells showed enrichment of terms related to RNA metabolic process and transcription. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of upregulated genes in SP cells revealed enrichment of signaling pathways associated with maintenance and differentiation of stem/progenitor cells. The terms involved in TCA cycles were enriched in GO and KEGG pathway analysis of downregulated genes in SP cells. These results support the assumption that bovine endometrial SP cells exhibit characteristics of somatic stem/progenitor cells. The ratio of SP cells to endometrial cells was lowest on days 9–11 after parturition, which gradually increased thereafter. SP cells were shown to differentiate into epithelial cells. Collectively, these results suggest that bovine endometrial SP cells were temporarily reduced immediately after calving possibly due to their differentiation to provide new endometrial cells.

DOI： 10.1093/biolre/ioab004

Neurokinin 3 receptor–selective agonist, senktide, decreases core temperature in Japanese Black cattle Reviewed

S. Nakamura, M. Miwa, Y. Morita, S. Ohkura, T. Yamamura, Y. Wakabayashi, S. Matsuyama

Domestic Animal Endocrinology Vol. 74 page： 106522 - 106522 2021.1

Seasonal changes in the reproductive performance in local cows receiving artificial insemination in the Pursat province of Cambodia. Reviewed International journal

Bengthay Tep, Yasuhiro Morita, Shuichi Matsuyama, Satoshi Ohkura, Naoko Inoue, Hiroko Tsukamura, Yoshihisa Uenoyama, Vutha Pheng

Asian-Australasian journal of animal sciences Vol. 33 ( 12 ) page： 1922 - 1929 2020.12

Language：Japanese Publishing type：Research paper (scientific journal)

Objective: The present study aimed to survey seasonal changes in reproductive performance in local cows receiving artificial insemination (AI) in the Pursat province of Cambodia, a tropical country, to investigate if ambient conditions affect the reproductive performance of cows as to better understand the major problems regarding cattle production. Methods: The number of cows receiving AI, resultant number of calving and calving rate were analyzed for those receiving the first AI from 2016 to 2017. The year was divided into three seasons: cool/dry (from November to February), hot/dry (from March to June), and wet (from July to October), based on the maximal temperature and rainfall in Pursat, to analyze the relationship between ambient conditions and the reproductive performance of cows. Body condition scores (BCS) and feeding schemes were also analyzed in these seasons. Results: The number of cows receiving AI was significantly higher in the cool/dry season than the wet season. The number of calving and calving rate were significantly higher in cows receiving AI in the cool/dry season compared with the hot/dry and wet seasons. The cows showed higher BCSs in the cool/dry season compared to the hot/dry and wet seasons probably due to the seasonal changes in the feeding schemes: These cows grazed on wild grasses in the cool/dry season but fed with a limited amount of grasses and straw in the hot/dry and wet seasons. Conclusion: The present study suggests that the low number of cows receiving AI, low number of calving and low calving rate could be mainly due to poor body condition as a result of the poor feeding schemes during the hot/dry and wet seasons. The improvement of body condition by the refinement of feeding schemes may contribute to an increase in the reproductive performance in cows during the hot/dry and wet seasons in Cambodia.

DOI： 10.5713/ajas.19.0893

Facilitatory and inhibitory role of central amylin administration in the regulation of the gonadotropin-releasing hormone pulse generator activity in goats. Reviewed International journal

Yuri Kitagawa, Takuya Sasaki, Reika Suzumura, Ai Morishima, Ryoki Tatebayashi, Assadullah, Nahoko Ieda, Yasuhiro Morita, Shuichi Matsuyama, Naoko Inoue, Yoshihisa Uenoyama, Hiroko Tsukamura, Satoshi Ohkura

Neuroscience letters Vol. 736 page： 135276 - 135276 2020.9

Language：Japanese Publishing type：Research paper (scientific journal)

Pulsatile gonadotropin-releasing hormone (GnRH) secretion is essential for regulating reproductive functions in mammals. GnRH pulses are governed by a neural mechanism that is termed the GnRH pulse generator. In the present study, we investigated the role of central calcitonin receptor (CTR) signaling in the regulation of the GnRH pulse generator activity in ovariectomized goats by administering amylin, an endogenous ligand for CTR, into the lateral ventricle. GnRH pulse generator activity was measured using multiple unit activity (MUA) recordings in the mediobasal hypothalamus. We analyzed changes in the interval of characteristic increases in MUA (MUA volleys). The MUA volley interval shortened immediately after amylin administration, followed by prolonged intervals. Double in situ hybridization for KISS1 (kisspeptin gene) and CALCR (CTR gene) revealed that low expression levels of CALCR were found in the arcuate kisspeptin neurons, which is suggested as the main population of neurons, involved in GnRH pulse generator activity. These results suggest that central amylin-CTR signaling has a biphasic role in the regulation of GnRH pulse generator activity by acting on cells other than the arcuate kisspeptin neurons in goats.

DOI： 10.1016/j.neulet.2020.135276

Peripheral administration of SB223412, a selective neurokinin-3 receptor antagonist, suppresses pulsatile luteinizing hormone secretion by acting on the gonadotropin-releasing hormone pulse generator in estrogen-treated ovariectomized female goats.

Takuya Sasaki, Tomoya Sonoda, Ryoki Tatebayashi, Yuri Kitagawa, Shinya Oishi, Koki Yamamoto, Nobutaka Fujii, Naoko Inoue, Yoshihisa Uenoyama, Hiroko Tsukamura, Kei-Ichiro Maeda, Fuko Matsuda, Yasuhiro Morita, Shuichi Matsuyama, Satoshi Ohkura

The Journal of reproduction and development Vol. 66 ( 4 ) page： 351 - 357 2020.8

Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development

Accumulating evidence suggests that KNDy neurons located in the hypothalamic arcuate nucleus (ARC), which are reported to express kisspeptin, neurokinin B, and dynorphin A, are indispensable for the gonadotropin-releasing hormone (GnRH) pulse generation that results in rhythmic GnRH secretion. The aims of the present study were to investigate the effects of peripheral administration of the neurokinin 3 receptor (NK3R/TACR3, a receptor for neurokinin B) antagonist, SB223412, on GnRH pulse-generating activity and pulsatile luteinizing hormone (LH) secretion in ovariectomized Shiba goats treated with luteal phase levels of estrogen. The NK3R antagonist was infused intravenously for 4 h {0.16 or 1.6 mg/(kg body weight [BW]·4 h)} during which multiple unit activity (MUA) in the ARC was recorded, an electrophysiological technique commonly employed to monitor GnRH pulse generator activity. In a separate experiment, the NK3R antagonist (40 or 200 mg/[kg BW·day]) was administered orally for 7 days to determine whether the NK3R antagonist could modulate pulsatile LH secretion when administered via the oral route. Intravenous infusion of the NK3R antagonist significantly increased the interval of episodic bursts of MUA compared with that of the controls. Oral administration of the antagonist for 7 days also significantly prolonged the interpulse interval of LH pulses. The results of this study demonstrate that peripheral administration of an NK3R antagonist suppresses pulsatile LH secretion by acting on the GnRH pulse generator, suggesting that NK3R antagonist administration could be used to modulate reproductive functions in ruminants.

DOI： 10.1262/jrd.2019-145

Establishment of immortalised cell lines derived from female Shiba goat KNDy and GnRH neurones Reviewed

Yuta Suetomi, Ryoki Tatebayashi, Shuhei Sonoda, Arisa Munetomo, Shuichi Matsuyama, Naoko Inoue, Yoshihisa Uenoyama, Yukari Takeuchi, Hiroko Tsukamura, Satoshi Ohkura, Fuko Matsuda

JOURNAL OF NEUROENDOCRINOLOGY Vol. 32 ( 6 ) page： e12857 2020.6

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：WILEY

Kisspeptin plays a critical role in governing gonadotrophin-releasing hormone (GnRH)/gonadotrophin secretion and subsequent reproductive function in mammals. The hypothalamic arcuate nucleus (ARC) kisspeptin neurones, which co-express neurokinin B (NKB) and dynorphin A (Dyn) and are referred to as KNDy neurones, are considered to be involved in GnRH generation. The present study aimed to establish cell lines derived from goat KNDy and GnRH neurones. Primary-cultured cells of female Shiba goat foetal hypothalamic ARC and preoptic area (POA) tissues were immortalised with the infection of lentivirus containing the simian virus 40 large T-antigen gene. Clones of the immortalised cells were selected by the gene expression of a neuronal marker, and then the neurone-derived cell clones were further selected by the gene expression of KNDy or GnRH neurone markers. As a result, we obtained a KNDy neurone cell line (GA28) from the ARC, as well as two GnRH neurone cell lines (GP11 and GP31) from the POA. Immunocytochemistry revealed the expression of kisspeptin, NKB and Dyn in GA28 cells, as well as GnRH in GP11 and GP31 cells. GnRH secretion from GP11 and GP31 cells into the media was confirmed by an enzyme immunoassay. Moreover, kisspeptin challenge increased intracellular Ca2+ levels in subsets of both GP11 and GP31 cells. Kisspeptin mRNA expression in GA28 cells, which expressed the oestrogen receptor alpha gene, was significantly reduced by 17 beta-oestradiol treatment. Furthermore, the transcriptional core promoter and repressive regions of the goat NKB gene were detected using GA28 cells. In conclusion, we have established goat KNDy and GnRH neurone cell lines that could be used to analyse molecular and cellular mechanisms regulating KNDy and GnRH neurones in vitro, facilitating the clarification of reproductive neuroendocrine mechanisms in ruminants.

DOI： 10.1111/jne.12857

Colocalization of GPR120 and anterior pituitary hormone-producing cells in female Japanese Black cattle.

Sho Nakamura, Kohei Noda, Masafumi Miwa, Shiori Minabe, Teruki Hagiwara, Akira Hirasawa, Shuichi Matsuyama, Ryutaro Moriyama

The Journal of reproduction and development Vol. 66 ( 2 ) page： 135 - 141 2020.4

Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development

Negative energy balance in domestic animals suppresses their reproductive function. These animals commonly use long-chain fatty acids (LCFAs) from adipocytes as an energy source under states of malnutrition. The G-protein coupled receptor, GPR120, is a specific receptor for LCFAs, but its role in reproductive function remains unknown in domestic animals. The purpose of this study was to examine whether GPR120 is involved in the reproductive system of cattle. GPR120 mRNA expression was evaluated in brain, pituitary, and ovarian tissue samples by RT-PCR. GPR120 gene expression was detected with high intensity only in the anterior pituitary sample, and GPR120-immunoreactive cells were found in the anterior pituitary gland. Double immunohistochemistry of GPR120 in the anterior pituitary hormone-producing cells, such as gonadotropes, thyrotropes, lactotropes, somatotropes, and corticotropes, was performed to clarify the distribution of GPR120 in the anterior pituitary gland of ovariectomized heifers. Luteinizing hormone β subunit (LHβ)- and follicle-stimulating hormone β subunit (FSHβ)-immunoreactive cells demonstrated GPR120 immunoreactivity at 80.7% and 85.9%, respectively. Thyrotropes, lactotropes, somatotropes, and corticotropes coexpressed GPR120 at 21.1%, 5.4%, 13.6%, and 14.5%, respectively. In conclusion, the present study suggests that GPR120 in the anterior pituitary gland might mediate LCFA signaling to regulate gonadotrope functions, such as hormone secretion or production, in cattle.

DOI： 10.1262/jrd.2019-111

Establishment of long-term chronic recording technique of in vivo ovarian parenchymal temperature in Japanese Black cows. Reviewed

Yasuhiro Morita, Riho Ozaki, Akihisa Mukaiyama, Takuya Sasaki, Ryoki Tatebayashi, Ai Morishima, Yuri Kitagawa, Reika Suzumura, Ryoya Abe, Hiroko Tsukamura, Shuichi Matsuyama, Satoshi Ohkura

The Journal of reproduction and development Vol. 66 ( 3 ) page： 271 - 275 2020

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development

The reproductive performance of cattle can be suppressed by heat stress. Reproductive organ temperature, especially ovarian temperature, may affect follicle development and ovulation. The establishment of a technique for long-term measurement of ovarian temperature could prove useful in understanding the mechanisms underlying the temperature-dependent changes in follicular development and subsequent ovulation in cows. Here we report a novel method facilitating long-term and continuous recording of ovarian parenchymal temperature in cows. The method revealed that the ovarian temperature in the luteal phase was constantly maintained lower than the vaginal temperature, and that the diurnal temperature variation in the ovary was significantly greater than that in the vagina, suggesting that the ovaries may require a lower temperature than other organs to maintain their functions. This novel method could be used for the further understanding of ovarian functions during estrous cycles in cows.

DOI： 10.1262/jrd.2019-097

Prepartum change in ventral tail base surface temperature in beef cattle: comparison with vaginal temperature and behavior indices, and effect of ambient temperature

MIWA Masafumi, MATSUYAMA Shuichi, NAKAMURA Sho, NODA Kohei, SAKATANI Miki

Journal of Reproduction and Development Vol. 65 ( 6 ) page： 515 - 525 2019.12

Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development

<p> Prediction of parturition is essential for sustainable production in beef and dairy cattle, yet the present methods are limited by their high invasiveness and low utility. Here we compared prepartum changes in ventral tail base surface temperature (ST) with changes in vaginal temperature (VT) and behavioral indices. We analyzed 22 parturitions from 22 beef cows. Changes in daily values of ST, VT, and behavioral indices over the 7 days before parturition were investigated. Hourly values were calculated as the actual values minus the mean values for the same hour over a 3-day period, and the changes in hourly values over the 48 h before parturition were investigated. To test the effect of ambient temperature, tested cows were assigned to two season-groups based on the ambient temperature to which they were exposed (warm: n = 13; cool: n = 9), and the daily and hourly values of the indices were compared between seasons. A decrease in ST occurred approximately 30 h before parturition, which was similar to the time of the decrease in VT and earlier than the increase of behavioral indices. In addition, a unique fluctuation of ST observed in the last few hours before parturition indicates that ST could provide a sign for parturition not only in the long-term like VT, but also in the short-term like behavioral indices. Although ST was more sensitive to ambient temperature than VT or the behavioral indices, the day of parturition could be predicted from ST in both the warm and cool seasons.</p>

DOI： 10.1262/jrd.2019-087

夏季におけるウシの排卵不全の原因探索

向山晃永, 尾崎理穂, 舘林亮輝, 北川悠梨, 阿部良哉, 鈴村玲香, 松山秀一, 大蔵聡, 森田康広

動物臨床医学会年次大会プロシーディング Vol. 40回 ( 5 ) page： 20 - 21 2019.11

ウシ主席卵胞および次席卵胞の顆粒層細胞において発現する遺伝子の網羅的解析

大下雪奈, 渡辺雄貴, 春日崇, 中平陽子, 棟朝亜理紗, 木村康二, 松山秀一, 大蔵聡, 真方文絵, 松田二子

日本獣医学会学術集会講演要旨集 Vol. 162回 page： 439 - 439 2019.8

Characterisation of bovine embryos following prolonged culture in embryonic stem cell medium containing leukaemia inhibitory factor. Reviewed International journal

Misa Hosoe, Tadashi Furusawa, Ken-Go Hayashi, Toru Takahashi, Yutaka Hashiyada, Keiichiro Kizaki, Kazuyoshi Hashizume, Tomoyuki Tokunaga, Shuichi Matsuyama, Ryosuke Sakumoto

Reproduction, fertility, and development Vol. 31 ( 6 ) page： 1157 - 1165 2019.5

Language：Japanese Publishing type：Research paper (scientific journal)

In order to help elucidate the process of epiblast and trophoblast cell differentiation in bovine embryos invitro, we attempted to develop a suitable culture medium to allow extended embryo culture. Day 7 bovine blastocysts developed in conventional medium were cultured further in embryonic stem cell medium with or without leukaemia inhibitory factor (LIF) until Day 23. At Day 14, the expression of octamer-binding transcription factor 3/4 (OCT3/4) and VIMENTIN was significantly higher in embryos cultured with than without LIF, but embryonic disc formation was not observed. Although expression of SRY (sex determining region Y)-box 17 (SOX17) mRNA was significantly lower in Day 14 embryos cultured with and without LIF than in invivo embryos, hypoblast cells formed just inside the trophoblast cells of the invitro-cultured embryos. On Day 23, expression of placental lactogen (PL) and prolactin-related protein 1 (PRP1) was not affected by LIF in invitro-cultured embryos, levels of both genes were significantly lower in the invitro than invivo embryos. Similar to invivo embryos, binucleate cell clusters seen in Day 23invitro-cultured embryos were composed of PL-negative and -positive cells. These results suggest that our culture system partially reproduced the differentiation process of trophoblast cells invivo.

DOI： 10.1071/RD18343

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