Papers - IWATA Hisataka
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A polysaccharide gel made of gellan gum improves oocyte maturation and embryonic development in pigs Reviewed International journal
Hara S, Shirasuna K, Iwata H.
J Reprod Dev. 70 ( 5 ) 303 - 308 2024.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：J-STAGE
Gellan gum (GG) is a soft, tractable, and natural polysaccharide substrate. Using the gel as substrate of oocyte in vitro maturation improves embryonic development via its antioxidant property.
DOI： 10.1262/jrd.2024-017
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Resveratrol intake by males increased the mitochondrial DNA copy number and telomere length of blastocysts derived from aged mice. Reviewed International journal
Teramoto N, Okada Y, Aburada N, Hayashi M, Ito J, Shirasuna K, Iwata H
J Reprod Dev. 70 ( 4 ) 247 - 252 2024.08
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
Telomere length of embryos shortens as male mice age. Present study shows that resveratrol intake ameliorates the age associated shorten in telomere length and decrease in mitochondrial DNA copy number in embryos through activation of SIRT1.
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Acetic acid affects porcine oocyte metabolism and improves oocyte developmental ability. Reviewed International journal
Inoue, Y., Hayashi, M., Shirasuna, K., & Iwata, H.
Theriogenology 224 174 - 182 2024.08
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier
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Telomere length determines the mitochondrial copy number in blastocyst-stage embryos. Reviewed International journal
Inoue, Y., Aoki, S., Ito, J., Hara, S., Shirasuna, K., & Iwata, H.
Mitochondrion 77 101877 2024.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier
We showed that Telomere length determines mitochondrial DNA copy number in embryos. Furthermore pathways associated with telomere length is predicted by RNAseq.
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Effect of low ethanol concentration in maturation medium on developmental ability, mitochondria, and gene expression profile in mouse oocytes. Reviewed International journal
Kizasu, S., Sato, T., Inoue, Y., Tasaki, H., Shirasuna, K., Okiishi, Y., & Iwata, H.
Reproductive biology 24 ( 2 ) 100854 2024.06
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier
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miRNAs in Follicular and Oviductal Fluids Support Global DNA Demethylation in Early-Stage Embryos Invited Reviewed International journal
Aoki, S., Inoue, Y., Hamazaki, M., Hara, S., Noguchi, T., Shirasuna, K., & Iwata, H.
International journal of molecular sciences 25 ( 11 ) 5872 2024.05
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：MDPI
miRNAs found in both oviductal fluid and follicular fluid induce demethylation of embryos until zygotic genome activation and improves embryonic development resulting in low methylated blastocysts.
DOI： https://doi.org/10.3390/ijms25115872
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microRNAs associated with the quality of follicular fluids affect oocyte and early embryonic development Reviewed International journal
Aoki S, Inoue Y, Hara S, Itou J, Shirasuna K, Iwata H.
Reproductive Medicine and Biology 23 ( 1 ) e12559 2024.01
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Reprod Med Biol.
miRNAs in the follicular fluid associated with oocytes developmental competence are defined. Supplementation of either oocyte maturation medium or embryo culture medium with specific miRNA improve embryonic development.
DOI： 10.1002/rmb2.12559.
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Effect of paternal aging and vitrification on mitochondrial DNA copy number and telomere length of mouse blastocysts International journal
ABURADA Nao, ITO Jun, INOUE Yuki, YAMAMOTO Taiyo, HAYASHI Masamune, TERAMOTO Noko, OKADA Yuri, KOSHIISHI Yuichi, SHIRASUNA Koumei, IWATA Hisataka
Journal of Reproduction and Development advpub ( 0 ) 65 - 71 2024
Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
<p>In this study, we examined the effects of paternal aging on the mitochondrial DNA copy number (mt-cn), telomere length (TL), and gene expression in mouse embryos. The effects of vitrification on the mt-cn and TL of the embryos derived from young and aged male parents (YF and AF, respectively) were examined. C57BL/6N male mice were used for embryo production at 13–23 and 50–55 weeks of age. Two-cell stage embryos were collected from the oviducts of superovulated female mice (8–15 weeks old) and cultured for 24 h until the 8-cell stage, followed by embryo vitrification. Fresh and vitrified-warmed embryos were incubated for 2 days until the blastocyst stage, and mt-cn and TL were investigated. The cell-free mitochondrial DNA copy number (cf-mt-cn) in the spent culture medium (SCM) of the embryos was then investigated. RNA sequencing of blastocysts revealed that metabolic pathways, including oxidative phosphorylation and mTOR pathways, were enriched in differentially expressed genes. The mt-cn and TL of AF-derived blastocysts were lower and shorter, respectively, than those of YF-derived blastocysts. Paternal aging did not affect the blastocyst rate after vitrification. Vitrification of the 8-cell stage embryos did not affect the mt-cn of the blastocysts. However, it increased the cf-mt-cn (cell-free mt-cn) in the SCM of both YF- and AF-derived embryos. Vitrification did not affect the TL of either YF- or AF-derived embryos. Thus, paternal aging affected the mt-cn and TL of the embryos, but vitrification did not affect these parameters in either age groups.</p>
DOI： 10.1262/jrd.2023-079
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A polysaccharide gel made of gellan gum improves oocyte maturation and embryonic development in pigs Reviewed International journal
HARA Shunsuke, SHIRASUNA Koumei, IWATA Hisataka
Journal of Reproduction and Development advpub ( 0 ) 2024
Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
<p>Gellan gum (GG) is a soft, tractable, and natural polysaccharide substrate used for cell incubation. In this study, we examined the effects of GG on porcine oocyte maturation. Cumulus cells and oocyte complexes (COCs) were collected from slaughterhouse-derived porcine ovaries and cultured on plastic plates containing 0.05% or 0.1% GG gels. The 0.1% GG gel improved the maturation rate and quality of blastocysts, as determined by the total cell number and the rate of abnormally condensed nuclei. GG gels have antioxidant abilities and oocytes cultured on GG gels (0.05% and 0.1%) have reduced reactive oxygen species (ROS) content. Furthermore, GG gels (0.05% and 0.1%) increased F-actin formation, whereas treatment of oocytes with H<sub>2</sub>O<sub>2</sub> reduced F-actin levels. GG gels increased the ATP content in oocytes but did not affect the mitochondrial DNA copy number or mitochondrial membrane potential. In addition, the medium cultured on 0.05% GG increased the glucose consumption of COCs. In conclusion, GG gel reduced ROS content, increased energy content, and improved subsequent embryonic development in pigs.</p>
DOI： 10.1262/jrd.2024-017
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Resveratrol intake by males increased the mitochondrial DNA copy number and telomere length of blastocysts derived from aged mice Reviewed International coauthorship International journal
TERAMOTO Noko, OKADA Yuri, ABURADA Nao, HAYASHI Masamune, ITO Jun, SHIRASUNA Komei, IWATA Hisataka
Journal of Reproduction and Development 70 ( 4 ) 247 - 253 2024
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
<p> The present study examined whether male resveratrol intake affected mitochondrial DNA copy number (mt-cn) and telomere length (TL) in blastocysts fathered by young and aged male mice. C57BL/6N male mice supplied with water or water containing 0.1 mM resveratrol were used for embryo production at 14–23 and 48–58 weeks of age. Two-cell-stage embryos were collected from the oviducts of superovulated female mice (8–15 weeks old) and cultured for 3 days until the blastocyst stage. Mt-cn and TL levels were measured by real-time polymerase chain reaction. Resveratrol intake did not affect body weight or water consumption. Resveratrol intake increased the expression levels of SIRT1 in the liver, the antioxidative ability of serum, and extended TL in the heart, whereas there was no significant difference in mt-cn in the heart or TL in sperm. The rate of blastocyst development was significantly lower in aged male mice than in younger mice, and resveratrol intake increased the total number of blastocysts derived from both young and aged males. Resveratrol intake did not affect mt-cn or TL in blastomeres of blastocyst-stage embryos derived from young mice, but significantly increased both mt-cn and TL in blastomeres of blastocysts derived from aged fathers. In conclusion, resveratrol intake increased mt-cn and TL levels in blastocysts derived from aged male mice.</p>
DOI： 10.1262/jrd.2024-043
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Age-related advanced glycation end-product accumulation impairs mitochondrial regulation after vitrification Reviewed International journal
Ito J, Iwata H.
Biology of Reproduction 109 ( 3 ) 271 - 281 2023.09
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Oxford Academic
Maternal age associated accumulation of advanced glycation end-products in oocytes hampers mitochondrial recovery following vitrification and activation of SIRT1 ameliorates the aged associated changes.
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ffect of a low ethanol concentration during in vitro maturation of bovine oocytes and subsequent embryo development. Reviewed International journal
Sato T, Hamazaki M, Inoue Y, Aoki S, Koshiishi Y, Shirasuna K, Iwata H.
Theriogenology 208 158 - 164 2023.09
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier
Low ethanol affects expression of genes regarding mitochondria in oocytes and alter mitochondrial function and quantity and histone modification in bovine embryos.
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Effect of a low ethanol concentration during in vitro maturation of bovine oocytes and subsequent embryo development. Reviewed International journal
208 158 - 164 2023.07
Language：English Publishing type：Research paper (scientific journal)
低濃度のエタノールを体外成熟培地に添加するとウシ胚のミトコンドリアが影響を受け、卵子の質が変わる。マウスとは異なり胚盤胞への発生得率が向上する。
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Age-related advanced glycation end-product accumulation impairs mitochondrial regulation after vitrification Reviewed International journal
Ito J, Iwata H.
Biology of Reproduction 2023.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
Age-associated accumulation of advanced glycation end-products in embryos hampers proper activation of mitochondrial quality control which in turn reduces mitochondrial quality in vitrified-warmed bovine embryos.
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Beneficial Effect of Polysaccharide Gel Made of Xanthan Gum and Locust Bean Gum on Bovine Oocyte Invited Reviewed International journal
Hara S, Inoue Y, Aoki S, Tanaka K, Shirasuna K, Iwata H.
International Journal of Molecular Science 24 ( 4 ) 3508 2023.02
Language：English
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Paternal aging impacts mitochondrial DNA content and telomere length in mouse embryos. Reviewed International journal
Ito J, Kageyama M, Hara S, Sato T, Shirasuna K, Iwata H.
Mitochondrion. 68 105 - 113 2023.01
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal)
Mitochondrial DNA (mtDNA) copy number and telomere length (TL) in blastocysts derived from the same male mice at young (10-19-week-old) and aged (40-49-week-old) time points and mtDNA and TL in the hearts of offspring derived from young and aged male mice were examined. Paternal aging correlated with reduced mtDNA and TL in blastocysts. mtDNA and TL were significantly correlated, which was also observed in bovine blastocysts. Moreover, mtDNA in the heart of offspring was reduced in male mice with paternal aging. In conclusion, paternal aging affects embryonic mtDNA and TL, potentially impacting their offspring.
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Age-associated changes in miRNA profile of bovine follicular fluid. Reviewed International journal
Nagata, S., Inoue, Y., Sato, T., Tanaka, K., Shinozawa, A., Shirasuna, K., & Iwata, H.
Reproduction 164 ( 5 ) 195 - 206 2022.10
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal)
We examined age-associated changes in miRNA profiles in the follicular fluid (FF) of cows. The role of miR-19b, which is abundant in the FF of young cows, in in vitro growth of early antral follicles (EAFs)-derived oocytes was assessed. FF was collected from the antral follicles of young (20-40 months) and aged (>120 months) cows. The miRNA profiles were similar between the FF of both age groups, whereas the abundance of some miRNAs differed between these samples. The miRNA profiles in granulosa cells (GCs) and the spent culture medium of oocyte-GC complexes (OGCs) derived from EAFs were distinct. Some miRNA groups overlapped among the GCs, culture media, and FFs. miR-19b was highly abundant in the FF of young cows, GCs, and culture medium. The supplementation of OGC culture medium with miR-19b increased the diameter, acetylation levels, and fertilisation ability of the oocytes. To assess whether miR-19b was functional in the GCs, a dual-luciferase assay, suppression of target protein, and RNA-sequencing of the GCs followed by functional annotation of the differentially expressed genes (DEGs) were conducted. Functional annotation of the DEGs suggested that miR-19b influences genes associated with FoxO signalling, endocytosis, and NR3C1 in GCs. These results suggest that in FFs, ageing affects the abundance of miRNAs that have important roles in oocyte development.
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Predicting of molecules mediating an interaction between bovine embryos and uterine epithelial cells. Reviewed International journal
Noguchi T, Hayashi T, Inoue Y, Hara S, Shirasuna K, Iwata H.
The Journal of reproduction and development 68 ( 5 ) 318 - 323 2022.10
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal)
Embryo-maternal reproductive tract interactions are pivotal for successful pregnancy. The present study predicted the molecules modulating embryo-uterine communication by comparing two sets of differentially expressed genes (DEGs): DEGs in uterine epithelial cells (UECs) collected from the uterus with and without blastocysts and DEGs between blastocysts developed in vivo and in vitro. Cows were subjected to super ovulation (SOV), followed by insemination or non-insemination at estrus (SOV + AI and SOV cows). Seven days after estrus, the uterus was flushed to collect UECs, and the presence of blastocysts in the uterus was confirmed. UECs were subjected to RNA-Sequencing (RNA-Seq) to identify DEGs. Publicly available RNA-Seq data of in vivo and in vitro developed bovine blastocysts were used to determine DEGs. Then, using ingenuity pathway analysis, activated- and inhibited-upstream regulators (USRs) for UECs in blastocysts were compared with those for blastocysts developed in vivo. RNA-Seq of UECs revealed that the DEGs were associated with immune response and cell adhesion pathways. The activated and inhibited USRs of UECs derived from SOV+ AI cows overlapped with the activated and inhibited USRs of blastocysts developed in vivo. Overlapping activated USRs include leukemia inhibitory factor, interleukin 6, fibroblast growth factor-2, transforming growth factor beta-1, and epidermal growth factor. In conclusion, the present study predicted the molecules that potentially mediate communication between the developing embryo and the uterus in vivo and prepare the uterus for pregnancy.
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miR-17-5p in bovine oviductal fluid affects embryo development. Reviewed International journal
Aoki S, Inoue Y, Shinozawa A, Tanaka K, Shirasuna K, Iwata H.
Mol Cell Endocrinol. 551 111651 2022.01
Authorship：Last author, Corresponding author Language：English Publishing type：Research paper (scientific journal)
This study identified microRNAs (miRNAs) in bovine oviductal fluids (OFs) and examined the effect of miR-17-5p in OFs on embryonic development to the blastocyst stage. Small RNA-seq of extracellular vesicles of OFs revealed 242 miRNAs. Additionally, analyzing expressions of randomly selected OF-miRNAs with RT-qPCR in the culture medium of oviductal epithelial cells indicated that the abundance of miRNAs in OFs increased during the luteal phase. miR-17-5p mimic-treated eight-cell-stage zona pellucida-free embryos showed improved embryonic development to the blastocyst stage. The effect of the miR-17-5p mimic was confirmed using a dual-luciferase assay and immunostaining. In addition, RNA-seq of the miR-17-5p mimic- or control-treated embryos revealed differentially expressed genes (DEGs), suggesting possible pathways that overlapped with the in silico-predicted pathways for miR-17-5p targeting genes. Furthermore, ingenuity pathway analysis of DEG predicted miR-17 to be a significant upstream regulator. Our results suggest that miR-17-5p in OFs regulates embryonic development in bovines.
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Predicting of molecules mediating an interaction between bovine embryos and uterine epithelial cells
NOGUCHI Tatsuo, HAYASHI Takeshi, INOUE Yuki, HARA Shunsuke, SHIRASUNA Koumei, IWATA Hisataka
Journal of Reproduction and Development 68 ( 5 ) 318 - 323 2022
Language：English Publisher：The Society for Reproduction and Development
<p> Embryo-maternal reproductive tract interactions are pivotal for successful pregnancy. The present study predicted the molecules modulating embryo-uterine communication by comparing two sets of differentially expressed genes (DEGs): DEGs in uterine epithelial cells (UECs) collected from the uterus with and without blastocysts and DEGs between blastocysts developed <i>in vivo</i> and <i>in vitro</i>. Cows were subjected to super ovulation (SOV), followed by insemination or non-insemination at estrus (SOV + AI and SOV cows). Seven days after estrus, the uterus was flushed to collect UECs, and the presence of blastocysts in the uterus was confirmed. UECs were subjected to RNA-Sequencing (RNA-Seq) to identify DEGs. Publicly available RNA-Seq data of <i>in vivo</i> and <i>in vitro</i> developed bovine blastocysts were used to determine DEGs. Then, using ingenuity pathway analysis, activated- and inhibited-upstream regulators (USRs) for UECs in blastocysts were compared with those for blastocysts developed <i>in vivo</i>. RNA-Seq of UECs revealed that the DEGs were associated with immune response and cell adhesion pathways. The activated and inhibited USRs of UECs derived from SOV+ AI cows overlapped with the activated and inhibited USRs of blastocysts developed <i>in vivo</i>. Overlapping activated USRs include leukemia inhibitory factor, interleukin 6, fibroblast growth factor-2, transforming growth factor beta-1, and epidermal growth factor. In conclusion, the present study predicted the molecules that potentially mediate communication between the developing embryo and the uterus <i>in vivo</i> and prepare the uterus for pregnancy.</p>
DOI： 10.1262/jrd.2022-046
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Effects of a gel culture system made of polysaccharides (xanthan gum and locust bean gum) on in vitro bovine oocyte development and gene expression of the granulosa cells. Reviewed
Sugimoto A, Inoue Y, Tanaka K, Sinozawa A, Shirasuna K, Iwata H
Molecular Reproduction and Development 88 ( 7 ) 516 - 524 2021.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1002/mrd.23518.
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Resveratrol enhanced mitochondrial recovery from cryopreservation-induced damages in oocytes and embryos Invited Reviewed
Hisataka Iwata
Reprod Med Biol 20 ( 4 ) 419 - 426 2021.06
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1002/rmb2.12401.
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Effect of maternal aging and vitrification on mitochondrial DNA copy number in embryos and spent culture medium. Reviewed
Aoki S, Ito J, Hara S, Shirasuna K, Iwata H.
Reproductive Biology 21 ( 2 ) 100506 2021.06
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Mitochondrial reactive oxygen species regulate mitochondrial biogenesis in porcine embryos Reviewed
Kageyama M, Ito J, Shirasuna K, Kuwayama T, Iwata H
Journal of Reproduction and Development 67 ( 2 ) 141 - 147 2021.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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凍結に由来するミトコンドリア障害からの胚の回復 Invited Reviewed
伊藤 洵、 岩田 尚孝
Journal of mammalian Ova Research 38 ( 1 ) 25 - 29 2021.04
Authorship：Lead author Language：Japanese Publishing type：Research paper (scientific journal)
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Effects of a gel culture system made of polysaccharides on in vitro bovine oocyte development
SUGIMOTO Ayaka, INOUE Yuuki, TANAKA Keisuke, IWATA Hisataka
The Journal of Reproduction and Development Supplement 114 ( 0 ) OR-5 - OR-5 2021
Language：Japanese Publisher：The Society for Reproduction and Development
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Substrate made of polysaccharides improves bovine oocyte maturation and subsequent embryo development
HARA Shunsuke, INOUE Yuki, TANAKA Keisuke, NOGUCHI Tatsuo, SIRASUNA Koumei, IWATA Hisataka
The Journal of Reproduction and Development Supplement 114 ( 0 ) OR-6 - OR-6 2021
Language：Japanese Publisher：The Society for Reproduction and Development
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Age associated change of miRNA in bovine follicular fluids and effect of these miRNAs on growth of oocytes derived from early antral follicles.
NAGATA Shuuta, YAMAGUCHI Hitoki, INOUE Yuki, TANAKA Keisuke, SHIRASUNA Koumei, IWATA Hisataka
The Journal of Reproduction and Development Supplement 114 ( 0 ) OR-3 - OR-3 2021
Language：Japanese Publisher：The Society for Reproduction and Development
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miRNA in bovine oviductal fluid affect embryo development
AOKI Sogo, INOUE Yuki, ISHIDA Daiki, TANAKA Keisuke, NOGUCHI Tatsuo, SHIRASUNA Koumei, IWATA Hisataka
The Journal of Reproduction and Development Supplement 114 ( 0 ) AW-7 - AW-7 2021
Language：Japanese Publisher：The Society for Reproduction and Development
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Age associated dynamics of mitochondrial gene expression in oocyte, embryo, and granulosa cells determined by RNA-seq
INOUE Yuki, TANAKA Keisuke, IWATA Hisataka
The Journal of Reproduction and Development Supplement 114 ( 0 ) P-6 - P-6 2021
Language：Japanese Publisher：The Society for Reproduction and Development
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Mitochondrial reactive oxygen species regulate mitochondrial biogenesis in porcine embryos International journal
KAGEYAMA Mio, ITO Jun, SHIRASUNA Koumei, KUWAYAMA Takehito, IWATA Hisataka
Journal of Reproduction and Development 67 ( 2 ) 141 - 147 2021
Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
<p> The number of mitochondria in blastocysts is a potential marker of embryo quality. However, the molecular mechanisms governing the mitochondrial number in embryos are unclear. This study was conducted to investigate the effect of reduced mitochondrial reactive oxygen species (ROS) levels on mitochondrial biogenesis in porcine embryos. Oocytes were collected from gilt ovaries and activated to generate over 4 cell-stage embryos at day 2 after activation. These embryos were cultured in media containing either 0.1 μM MitoTEMPOL (MitoT), 0.5 μM Mitoquinol (MitoQ), or vehicle (ethanol) for 5 days to determine the rate of development to the blastocyst stage. The mitochondrial number in blastocysts was evaluated by real-time polymerase chain reaction (PCR). Five days after activation, the embryos (early morula stage) were subjected to immunostaining to determine the expression levels of NRF2 in the nucleus. In addition, the expression levels of <i>PGC1α</i> and <i>TFAM</i> in the embryos were examined by reverse transcription PCR. One day of incubation with the antioxidants reduced the ROS content in the embryos but did not affect the rate of development to the blastocyst stage. Blastocysts developed in medium containing MitoT had lower mitochondrial DNA copy numbers and ATP content, whereas MitoQ showed similar but insignificantly trends. Treatment of embryos with either MitoT or MitoQ decreased the expression levels of NRF2 in the nucleus and levels of <i>PGC1α</i> and <i>TFAM</i>. These findings indicate that reductions in mitochondrial ROS levels are associated with low mitochondrial biogenesis in embryos.</p>
DOI： 10.1262/jrd.2020-111
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HIRATA Yoshiki, KATSUKURA Yusuke, HENMI Yuka, OZAWA Ren, SHIMAZAKI Sayaka, KUROSAWA Akira, TORII Yasushi, TAKAHASHI Hironori, IWATA Hisataka, KUWAYAMA Takehito, SHIRASUNA Koumei
Journal of Reproduction and Development 67 ( 4 ) 257 - 264 2021
Language：English Publisher：The Society for Reproduction and Development
<p> Advanced maternal age is a risk factor for female infertility, and placental dysfunction is considered one of the causes of pregnancy complications. We investigated the effects of advanced maternal aging on pregnancy outcomes and placental senescence. Female pregnant mice were separated into three groups: young (3 months old), middle (8–9 months old), and aged (11–13 months old). Although the body weights of young and middle dams gradually increased during pregnancy, the body weight of aged dams only increased slightly. The placental weight and resorption rate were significantly higher, and live fetal weights were reduced in a maternal age-dependent manner. Although mRNA expression of senescence regulatory factors (p16 and p21) increased in the spleen of aged dams, mRNA expression of p16 did not change and that of p21 was reduced in the placenta of aged dams. Using a cytokine array of proteins extracted from placental tissues, the expression of various types of senescence-associated secretory phenotype (SASP) factors was decreased in aged dams compared with young and middle dams. The aged maternal placenta showed reduced immune cell accumulation compared with the young placenta. Our present results suggest that models using pregnant mice older than 8 months are more suitable for verifying older human pregnancies. These findings suggest that general cellular senescence programs may not be included in the placenta and that placental functions, including SASP production and immune cell accumulation, gradually decrease in a maternal age-dependent manner, resulting in a higher rate of pregnancy complications.</p>
DOI： 10.1262/jrd.2021-034
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Comparative analysis of cell-free DNA content in culture medium and mitochondrial DNA copy number in porcine parthenogenetically activated embryos. Reviewed
Kobayashi M, Ito J, Shirasuna K, Kuwayama T, Iwata H.
Journal of Reproduction and Development 66 ( 6 ) 539 - 546 2020.12
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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ウシの分娩前後における血漿中ミトコンドリア由来細胞外DNA量の変化と血液成分値との関係 Reviewed
野口 龍生, 相澤 拓朗, 岩田 尚孝
日本畜産学会報 91 ( 3 ) 227 - 232 2020.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Prediction of major microRNAs in follicular fluid regulating porcine oocyte development Reviewed
Yuki Inoue, Yasuhisa Munakata, Akihisa Shinozawa, Ryoka Kawahara-Miki, Koumei Shirasuna, Hisataka Iwata
Journal of Assisted Reproduction and Genetics 37 ( 10 ) 2569 - 2579 2020.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Xanthan gum and locust bean gum substrate improves bovine embryo developmen Reviewed
Hara S, Aoki S, Nagata M, Shirasuna K, Noguchi T, Iwata H.
Reproduction in Domestic Animals. 55 ( 9 ) 1124 - 1131 2020.09
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1111/rda.13750.
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Cell free mitochondrial DNA in plasma and physical condition of cows
NOGUCHI Tatsuo, AIZAWA Takuro, IWATA Hisataka
Nihon Chikusan Gakkaiho 91 ( 3 ) 227 - 232 2020.08
Language：Japanese Publisher：Japanese Society of Animal Science
<p>Blood contains cell free DNA (cfDNA), and cell free mitochondrial DNA(cf-mtDNA) is able to be easily detected by real time PCR. The present study investigated the relationship between the amount of cf-mtDNA in plasma and physiological conditions of cows. In addition, relationship between preg-nancy following artificial insemination and concentration of cf-mtDNA in the corresponding plasma. Significantly positive relationship was found between concentration of LDH and cf-mtDNA, and negative relationship was found between concentration of triglyceride and cf-mtDNA. There was no significant difference in the amount of cf-mtDNA between pregnant or non-pregnant cows following artificial insemination.</p>
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Abundance of cell-free mitochondrial DNA in spent culture medium associated with morphokinetics and blastocyst collapse of expanded blastocysts. Reviewed
Kobayashi M, Kobayashi J, Shirasuna K, Iwata H
Reproductive Medicine and Biology 19 ( 4 ) 404 - 414 2020.08
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1002/rmb2.12344
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Comparison of Gene Expression and Mitochondria Number Between Bovine Blastocysts Obtained in Vitro and in Vivo Reviewed
Tatsuo Noguchi, Takuro Aizawa, Yasuhisa Munakata, Hisataka Iwata
Journal of Reproduction and Development 66 ( 1 ) 35 - 39 2020.02
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Resveratrol treatment increases mitochondrial biogenesis and improves viability of porcine germinal-vesicle stage vitrified-warmed oocytes Reviewed
Ito J, Shirasuna K, Kuwayama T, Iwata H.
Cryobiology 93 37 - 43 2020
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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SANO Michiya, SHIMAZAKI Sayaka, KANEKO Yasuaki, KARASAWA Tadayoshi, TAKAHASHI Masafumi, OHKUCHI Akihide, TAKAHASHI Hironori, KUROSAWA Akira, TORII Yasushi, IWATA Hisataka, KUWAYAMA Takehito, SHIRASUNA Koumei
Journal of Reproduction and Development 66 ( 3 ) 241 - 248 2020
Language：English Publisher：The Society for Reproduction and Development
<p> Maternal obesity is one of the major risk factors for pregnancy complications and is associated with low-grade chronic systemic inflammation due to higher levels of pro-inflammatory cytokines such as interleukin (IL)-1β. Pregnant women with obesity have abnormal lipid profiles, characterized by higher levels of free fatty acids, especially palmitic acid (PA). Previously, we reported that PA stimulated IL-1β secretion via activation of NLRP3 inflammasome in human placental cells. These observations led us to hypothesize that higher levels of PA induce NLRP3 inflammasome activation and placental inflammation, resulting in pregnancy complications. However, the effects of PA on NLRP3 inflammasome during pregnancy <i>in vivo</i> remain unclear. Therefore, PA solutions were administered intravenously into pregnant mice on day 12 of gestation. Maternal body weight was significantly decreased and absorption rates were significantly higher in PA-injected mice. The administration of PA significantly increased IL-1β protein and the mRNA expression of NLRP3 inflammasome components (<i>NLRP3</i>, <i>ASC</i>, and caspase-1) within the placenta. In murine placental cell culture, PA significantly stimulated IL-1β secretion, and this secretion was suppressed by a specific NLRP3 inhibitor (MCC950). Simultaneously, the number of macrophages/monocytes and neutrophils, together with the mRNA expression of these chemokines increased significantly in the placentas of PA-treated mice. Treatment with PA induced ASC assembling and IL-1β secretion in macrophages, and this PA-induced IL-1β secretion was significantly suppressed in NLRP3-knockdown macrophages. These results indicate that transient higher levels of PA exposure in pregnant mice activates NLRP3 inflammasome and induces placental inflammation, resulting in the incidence of absorption.</p>
DOI： 10.1262/jrd.2020-007
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Effect of aging on mitochondria and metabolism of bovine granulosa cells International journal
NAGATA Shuta, TATEMATSU Kaoru, KANSAKU Kazuki, INOUE Yuki, KOBAYASHI Mitsuru, SHIRASUNA Koumei, IWATA Hisataka
Journal of Reproduction and Development 66 ( 6 ) 547 - 554 2020
Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
<p> This study investigated the effect of aging on mitochondria in granulosa cells (GCs) collected from the antral follicles of young and aged cows (25–50 months and over 140 months in age, respectively). When GCs were cultured under 20% O<sub>2</sub> for 4 days, mitochondrial DNA copy number (Mt-number), determined by real-time PCR, increased throughout the culture period, and the extent of increase was greater in the GCs of young cows than in those of old cows. In a second experiment, GCs were cultured under 20% O<sub>2</sub> for 24 h. Protein levels of TOMM20 and TFAM in GCs were lower in aged cows than in young cows, and the amount of reactive oxygen species and the mitochondrial membrane potential were higher, whereas ATP content and proliferation activity were lower, respectively. Glucose consumption and lactate production were higher in the GCs of aged cows than in those of young cows. When GCs were cultured under 5% or 20% O<sub>2</sub> for 24 h, low O<sub>2</sub> decreased ATP content and increased glucose consumption in GCs of both age groups compared with high O<sub>2</sub>; however, low O<sub>2</sub> decreased the Mt-number only in the GCs of young cows. In conclusion, we show that aging affects mitochondrial quantity, function, and response to differential O<sub>2</sub> tensions in GCs.</p>
DOI： 10.1262/jrd.2020-071
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Comparison of gene expression and mitochondria number between bovine blastocysts obtained <i>in vitro</i> and <i>in vivo</i> International journal
NOGUCHI Tatsuo, AIZAWA Takuro, MUNAKATA Yasuhisa, IWATA Hisataka
Journal of Reproduction and Development 66 ( 1 ) 35 - 39 2020
Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
<p> Embryo transfer uses embryos developed <i>in vivo</i> or <i>in vitro</i> for cattle production, however there is a difference in the quality of the embryos obtained by the two methods. This study addresses the differences in gene expression between blastocysts developed <i>in vitro</i> and <i>in vivo</i>. <i>In vivo</i> blastocysts were flushed from the uteri of super-ovulated cows and blastocysts developed <i>in vitro</i> were derived from <i>in vitro</i> matured and fertilized embryos. The same batch of frozen bull sperm was used for insemination and <i>in vitro</i> fertilization. Blastocysts were then subjected to RNA sequencing. Differentially expressed genes upregulated in <i>in vitro</i> blastocysts were annotated to focal adhesion, extracellular matrix (ECM)-receptor interaction, and PI3K-Akt signaling and the genes that were upregulated in <i>in vivo</i> blastocysts were annotated to oxidation-reduction processes, mitochondrion organization, and mitochondrial translation. Although the total cell number of the two types of blastocysts was similar, the mitochondrial quantity (determined by mitochondrial DNA copy numbers and expression levels of TOMM20), and ATP content in the blastocysts were lower in <i>in vivo</i> blastocysts compared with those developed <i>in vitro</i>. In conclusion, RNAseq revealed differential molecular backgrounds between <i>in vitro</i> and <i>in vivo</i> developed blastocysts and mitochondrial number and function are responsible for these differences.</p>
DOI： 10.1262/jrd.2019-100
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Comparative analysis of cell-free DNA content in culture medium and mitochondrial DNA copy number in porcine parthenogenetically activated embryos International journal
KOBAYASHI Mitsuru, ITO Jun, SHIRASUNA Koumei, KUWAYAMA Takehito, IWATA Hisataka
Journal of Reproduction and Development 66 ( 6 ) 539 - 546 2020
Language：English Publishing type：Research paper (scientific journal) Publisher：The Society for Reproduction and Development
<p> We examined the effect of ploidy on mitochondrial DNA (mtDNA) copy number in embryos and the amount of cell-free mitochondrial and nucleic DNA content (cf-mtDNA and cf-nDNA) in spent culture medium (SCM). Oocytes collected from the ovaries were matured, activated, incubated in medium containing cycloheximide (CHX) or CHX and cytochalasin B (CB) for 4.5 h to produce haploid or diploid embryos (H-group and D-group embryos). These embryos were cultured for 7 days, and the blastocysts and SCM were examined. The amount of mtDNA and nDNA was determined by real-time PCR. The rate of development to the blastocyst stage was higher for the D-group than for the H-group. Moreover, D-group blastocysts had less mtDNA compared to the H-group blastocysts. After activation, the mitochondrial content was constant before the blastocyst stage in D-group embryos, but increased earlier in H-group embryos. The amount of cf-mtDNA in the SCM of D-group blastocysts was greater than that of H-group blastocysts. However, when the cf-mtDNA in the SCM of 2 cell-stage embryos (day 2 post-activation) was examined, the amount of cf-mtDNA was greater in the H-group than in the D-group embryos. When D-group embryos were cultured for 7 days, a significant correlation was observed between the total cell number of blastocysts and cf-nDNA content in the SCM. Hence, although careful consideration is needed regarding the time point for evaluating mtDNA content in the embryos and SCM, this study demonstrates that mtDNA in the embryos and SCM was affected by the ploidy of the embryos.</p>
DOI： 10.1262/jrd.2020-097
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Cell-free DNA Content in Follicular Fluid: A Marker for the Developmental Ability of Porcine Oocytes Reviewed
Kana Ichikawa, Hidenori Shibahara, Komei Shirasuna, Takehito Kuwayama, Hisataka Iwata
Reproductive Medicine and Biology 19 ( 1 ) 95 - 103 2019.12
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1002/rmb2.12309
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Xanthan Gum and Locust Bean Gum Gel Supports in Vitro Development of Porcine Oocytes Derived From Early Antral Follicles Reviewed
Yasuhisa Munakata, Ayaka Sugimoto, Koumei Shirasuna, Takehito Kuwayama, Hisataka Iwata
Journal of Reproduction and Development 65 ( 6 ) 551 - 554 2019.12
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1262/jrd.2019-074
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Mitochondrial Cell-free DNA secreted from porcine granulosa cells Reviewed
K Kazuki, Y Munakata, K Shirasuna, T Kuwayama, H Iwata
Zygote 27 ( 5 ) 272 - 278 2019.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Modifying the medium volume and gel substrate in in vitro culture conditions improves growth of porcine oocytes derived from early antral follicles. Reviewed
Shibahara H, Munakata Y, Ishiguro A, Koumei S, T Kuwayama, H Iwata
Journal of Reproduction and Development 65 ( 4 ) 375 - 379 2019.08
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Effects of resveratrol treatment on mitochondria and subsequent embryonic development of bovine blastocysts cryopreserved by slow freezing Reviewed
T Hayashi, K Kansaku, T Abe, S Ueda and H Iwata
Animal science Journal 90 ( 7 ) 849 - 856 2019.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1111/asj.13219.
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Cell-free DNA in medium is associated with the maturation ability of in vitro cultured oocytes Reviewed
Munakata Y, Shirasuna K, Kuwayama T, Iwata H.
journal of Reproduction and Development 65 ( 2 ) 171 - 175 2019.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Follicular factors determining the developmental competence of porcine oocyte Reviewed
H Shibahara, A Ishiguro, K Shirasuna, T Kuwayama, H Iwata
Reproductive Medicine and Biology 18 ( 3 ) 256 - 262 2019.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1002/rmb2.12269.
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Resveratrol enhances the clearance of mitochondrial damage by vitrification and Reviewed
Hara T, Kin A, Aoki S, Nakamura S, Shirasuna K, Kuwayama T, Iwata H.
PLoS One. 13 ( 10 ) e0204571 2018.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Follicular factors determining granulosa cell number and Reviewed
Munakata Y, Ueda M, Kawahara-Miki R, Kansaku K, Itami N, Shirasuna K, Kuwayama
J Assist Reprod Genet 35 ( 10 ) 1809 - 1819 2018.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Palmitic acid induces ceramide accumulation, mitochondrial protein hyperacetylation, and mitochondrial dysfunction in porcine oocytes. Reviewed
Itami N, Shirasuna K, Kuwayama T, Iwata H.
Biology of Reproduction 98 ( 5 ) 644 - 633 2018.05
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Short-term heat stress induces mitochondrial degradation and biogenesis and enhances mitochondrial quality in Reviewed
Itami N, Shirasuna K, Kuwayama T, Iwata H.
Journal of Thermal Biology 74 256 - 263 2018.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Influence of resveratrol pretreatment on thawed bovine embryo quality and mitochondrial DNA copy number. Reviewed
Hayashi T, Ueda S, Mori M, Baba T, Abe T, Iwata H.
Theriogenology 106 271 - 278 2018.01
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Non-essential fatty acid-associated ability of follicular fluid to support porcine oocyte maturation and development Reviewed
Ogawa K, Itami N, Ueda M, Kansaku K, Shirasuna K, Kuwayama T, Iwata H.
Reproductive Medicine andBiology 17 ( 2 ) 155 - 163 2018.01
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1002/rmb2.12084.
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Mitochondrial dysfunction in cumulus-oocyte complexes increases cell-free mitochondrial DNA. Reviewed
Kansaku K, Munakata Y, Itami N, Shirasuna K, Kuwayama T, Iwata H.
Journal of Reproduction and Development 64 ( 3 ) 261 - 266 2018
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1262/jrd.2018-012
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Maternal aging affects oocyte resilience to carbonyl cyanide-m-chlorophenylhydrazone -induced mitochondrial dysfunction in cows. Reviewed
Kansaku K, Takeo S, Itami N, Kin A, Shirasuna K, Kuwayama T, Iwata H.
PLos One 12 ( 11 ) e0188099 2017.11
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Differential effects of mitochondrial inhibitors on porcine granulosa cells and oocytes Reviewed
Kansaku K, Itami N, Kawahara-Miki R, Shirasuna K, Kuwayama T, Iwata H
Theriogenology 103 98 - 103 2017.11
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Modification of mitochondrial function, cytoplasmic lipid ontent and cryosensitivity of bovine embryos by resveratrol Reviewed
Abe T, Kawahara-Miki R, Hara T, Noguchi T, Hayashi T, Shirasuna K, Kuwayama T, Iwata H.
Journal of Reproduction and Development 63 ( 5 ) 455 - 461 2017.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Age-associated changes in granulosa cells and follicular fluid in cows. Invited Reviewed
Hisataka Iwata
Journal of Reproduction and Development 63 ( 4 ) 339 - 345 2017.08
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
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Age-associated deterioration in follicular fluid induces a decline in bovine oocyte quality Reviewed
Takeo S, Kimura K, Shirasuna K, Kuwayama T, Iwata H
Reproduction Fertility and Development 29 ( 4 ) 759 - 767 2017.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Promotion of glucose utilization by insulin enhances granulosa cell proliferation and developmental competence of porcine oocyte grown in vitro Reviewed
Itami N, Munakata Y, Shirasuna K, Kuwayama T, Iwata H.
Zygote 25 ( 1 ) 65 - 74 2017.02
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Polyacrylamide gel as a culture substrate improves in vitro oocyte growth from porcine early Reviewed
Munakata Y, Kawahara-Miki R, Shirasuna K, Kuwayama T, Iwata H.
Molecular Reproduction and Development 84 ( 1 ) 44 - 54 2017.01
Language：English Publishing type：Research paper (scientific journal)
DOI： 10.1002/mrd.22758.
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Low oxygen Reviewed
Shiratsuki S, Hara T, Munakata Y, Shirasuna K, Kuwayama T, Iwata H.
Molecular and Cellular Endocrinology 437 75 - 85 2016.12
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
顆粒層細胞が卵胞峰内で増殖するメカニズムはHIF1からVEGF,AKT-mTORの経路を介していることを明らかにした。
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Addition of granulosa cell mass to the culture medium of oocytes derived from early antral follicles increases oocyte growth, ATP content, and acetylation of H4K12. Reviewed
Sugiyama M, Sumiya M, Shirasuna K, Kuwayama T, Iwata H.
Zygote 24 ( 6 ) 848 - 856 2016.12
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Relationship between the number of cells surrounding oocytes and energy Reviewed
Munakata Y, Ichinose T, Ogawa K, Itami N, Tasaki H, Shirasuna K, Kuwayama T,Iwata H
Theriogenology 89 ( 7 ) 1789 - 1798 2016.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
卵子を取り巻く細胞の数が卵子の脂質含量、ATPそしてアセチル化を決定していることを明らかにした。
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Gene expression of granulosa cells and oocytes in various stages of follicle and in vitro grown oocytes and granulosa cell complexes Reviewed
Munakata Y, R Kawahara-Miki, S Shiratuski, H Tasaki, N Itami, K Shirasuna, T Kuwayama, H Iwata
Journal of Reproduction and Development 62 ( 4 ) 359 - 366 2016.08
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Age-associated events in bovine oocytes and possible countermeasures Invited Reviewed
Hisataka Iwata
Reproductive medicine and Biology 15 ( 3 ) 155 - 164 2016.07
Language：English Publishing type：Research paper (scientific journal)
加齢に伴う卵子の変化とその対処方法について概説した
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Maternal aging and mitochondria in oocytes Invited Reviewed
23 ( 2 ) 141 - 146 2016.06
Language：Japanese Publishing type：Research paper (scientific journal)
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Liver condition of Holstein cows affects mitochondrial function and fertilization ability of oocytes. Reviewed
Tanaka H, Takeo S, Abe T, Kin A, Shirasuna K, Kuwayama T, Iwata H
Journal of Reproduction and Development 62 ( 3 ) 235 - 240 2016.01
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
肝臓の疾患が卵子のミトコンドリアと受精率に及ぼす影響について明らかにした
DOI： 10.1071/RD15228
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ウシ卵巣中の卵胞数と加齢の関係 Reviewed
伊丹暢彦・岩田尚孝
日本胚移植学雑誌 37 ( 3 ) 113 - 117 2015.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
加齢に伴うウシの卵子数の減少・小卵胞の質の低下について紹介した
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加齢がウシ卵子の発育能力とミトコンドリアに及ぼす影響 Invited Reviewed
白築章互・岩田尚孝
日本胚移植学雑誌 37 ( 3 ) 119 - 125 2015.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
加齢に伴うウシの卵子のミトコンドリアの質の減退とレスベラトロールを用いた対処方法について紹介した
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加齢ウシの卵胞液の性状と卵子の体外成熟 Invited Reviewed
竹尾駿 ・岩田尚孝
日本胚移植雑誌 37 ( 3 ) 127 - 131 2015.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
加齢に伴うウシの卵胞液の性状変化と卵子の質低下に及ぼす影響について紹介した
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Mitochondrial biogenesis and degradation are induced by CCCP treatment of porcine oocytes. Reviewed
Itami N, Shiratsuki S, Shirasuna K, Kuwayama T, Iwata H.
Reproduction 150 ( 2 ) 97 - 104 2015.05
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
ミトコンドリアを人為的に機能不全にすると卵子中では新たなミトコンドリアの合成と分解が起こり卵子の能力が回復することを示した。
DOI： 10.1530/REP-15-0037.
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Resveratrol improves the quality of pig oocytes derived from early antral follicles through SIRT1 activation Reviewed
N Itami, K Shirasuna, T Kuwayama, H Iwata
Theriogenology 83 ( 8 ) 1360 - 1367 2015.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
レスベラトロールはブタの小卵胞の発育を改善し、卵子中に蓄える脂の量を減らす。また卵子のミトコンドリアの機能を改善する
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Effect of 5-aminoimidazole-4-carboxamide-ribonucleoside on the mitochondrial function and developmental ability of bovine oocytes Reviewed
Shun Takeo, Takahito Abe, Koumei Shirasuna, Takehito Kuwayama, Hisataka Iwata
Theriogenology 84 ( 4 ) 490 - 497 2015.03
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
AICARで成熟中の卵子のAMPKを活性化するとミトコンドリアの機能がが改善しウシ卵子の能力が改善することを示した
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Effects of reaggregated of granulosa cells and oocytes derived from early antral follicles on the properties of oocytes grown in vitro Reviewed
Ayano Oi, Hidetaka Tasaki, Yasuhisa Munakata, Koumei Shirasuna, Takehito Kuwayama, and Hisataka Iwata
Journal of Reproduction and Development 61 ( 3 ) 191 - 197 2015.02
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
ブタの初期胞状卵胞を用いて細胞と顆粒層細胞をいったん剥離してもその後共培養することで卵子卵丘細胞複合体を再形成し卵子を体外発育させることができることを示した。
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Age-associated changes in bovine oocytes and granulosa cell complexes collected from early antral follicles. Reviewed
Itami N, Kawahara-Miki R, Kawana H, Endo M, Kuwayama T, Iwata H
J Assist Reprod Genet 2014.05
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
加齢ウシの初期胞状卵胞の顆粒層細胞の遺伝子発現プロファイルはすでに退行様になっているために体外発生能力が若齢のそれに比べて低いことを示した。
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培養皿の形状がウシ初期胞状卵胞由来卵子の Reviewed
川名宏典・松本美保・門司恭典・桑山岳人・岩田尚孝
日本胚移植雑誌 36 ( 2 ) 65 - 70 2014.05
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
培養皿底面の形状が球体の場合、従来の平面に比べてウシの初期胞状卵胞の体外培養成績が良好であることを示した。
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Relationship between mitochondrial DNA Copy Number and SIRT1 Expression in Porcine Oocytes. Reviewed
Sato D, Itami N, Tasaki H, Takeo S, Kuwayama T, Iwata H.
PlosOne 18 ( 9 ) e94488 - e94488 2014.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
レスベラトロールが卵子内部のミトコンドリアの更新を促し卵子の質を改善することを示した。またミトコンドリアがプロテアソームを介して除去されていることを示した。
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Resveratrol improves the mitochondrial function and fertilization outcome of bovine oocytes. Reviewed
Takeo S, Sato D, Kimura K, Monji Y, Kuwayama T, Kawahara-Miki R, Iwata H.
Journal of Reproduction and Development 60 ( 2 ) 92 - 99 2014.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
レスベラトロールがウシ卵子のミトコンドリアの品質を改善し発生率を向上させることを示した。
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Brilliant cresyl blue staining negatively affects mitochondrial functions in porcine oocytes. Reviewed
Santos EC, Sato D, Lucia T, Iwata H.
Zygote 20 1 - 8 2013.12
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
Brilliant Cresyl Blueは良好な卵子を選別するのに用いられるがこの試薬自体がミトコンドリアに酸化ストレスを与え活性酸素の産生を促し卵子の質を損なうことを示した。
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Supplementation of maturation medium with folic acid affects DNA methylation of porcine oocytes and histone acetylation of early developmental stage embryos Reviewed
D Sato, K Sakurai, Y Monji, T Kuwayama, H Iwata
Journal of Mammalian Ova reserch 30 ( 3 ) 109 - 116 2013.11
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
成熟培地への葉酸の添加がDNAのメチル化状態を変えゲノムの活性化時期のヒストンのアセチル化状態や発生率を変えることを示した
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Maternal liver damage delays meiotic resumption in bovine oocytes through impairment of signalling cascades originated from low p38MAPK activity in cumulus cells
H. Tanaka, S. Takeo, Y. Monji, T. Kuwayama, H. Iwata
Reproduction in Domestic Animals 2013.09
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
肝臓に疾患を有するウシの顆粒層細胞ではP38MAPKの活性が低くこれが減数分裂の進行を遅らせている要因であることを示した
DOI： 10.1111/rda.12235
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Liver condition affects bovine oocyte qualities by changing the characteristics of follicular fluid and plasma Reviewed
H. Tanaka1, K. Shibano2, Y. Monji1, T. Kuwayama1, H. Iwata
Reprod Domest Anim. 48 ( 4 ) 619 - 626 2013.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
肝臓の状態によって変化する血液性状に伴い卵胞駅の性状も変化することを示した また卵胞液中の増殖因子も減ることを示した
DOI： 10.1111/rda.12135.
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Effect of Maternal Age on the Ratio of Cleavage and Mitochondrial DNA Copy Number in Early Developmental Stage Bovine Embryos Reviewed
S Takeo, H Goto, T Kuwayama, Y Monji, H Iwata
Journal of Reproduction and Development. 59 ( 2 ) 174 - 179 2013.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
加齢により初期胚の分割率及びミトコンドリア数が減少することを示した。
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Age-Associated Changes in Gene Expression and Developmental Competence of Bovine Oocytes and a Possible Countermeasure Against Age-associated Events Reviewed
S. Takeo1, R. Kawahara-Miki2, H. Goto1, F. Cao3, K. Kimura, Y. Monji1, T. Kuwayama1, AND H.Iwata
Mol Reprod Dev. 80 ( 7 ) 508 - 521 2013.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
加齢に由来する卵子の異常を示し、SIRT1の活性化がこれを抑制するを示すとともに、加齢卵子の遺伝子発現を次世代シーケンサーを用いて網羅的に解析した。
DOI： 10.1002/mrd.22187.
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Estradiol has a major role in antrum formation of porcine preantral follicles cultured in vitro Reviewed
H Tasaki, H Iwata*, D Sato, Y Monji, and T Kuwayama
Theriogenology 79 ( 5 ) 809 - 814 2013.05
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
豚の初期胞状卵胞卵子ではエストラジールへの暴露が腔形成の引き金になっている可能性を示した。
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Estradiol supports in vitro development of bovine early antral follicles. Reviewed
Endo M, Kawahara-Miki R, Cao F, Kimura K, Kuwayama T, Monji Y, Iwata H
Reproduction 145 ( 1 ) 85 - 96 2013.01
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
牛の初期胞状卵胞卵子においてエストラジオールが正常な体外での卵胞発育に必要であることを遺伝子の網羅解析および培養試験において示した。
DOI： 10.1530/REP-12-0319
Other Link： http://www.reproduction-online.org/content/145/1/85.long
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Factors affecting the primary sex ratio in cows. Invited Reviewed
Hisataka Iwata
Journal of Mammalian Ova Research, 29 ( 1 ) 95 - 102 2012.10
Language：English Publishing type：Research paper (scientific journal)
人工授精や胚移植において産子の性の比率否が偏る現象とその原因を総説した
DOI： http://dx.doi.org/10.1274/jmor.29.96
Other Link： http://www.bioone.org/doi/full/10.1274/jmor.29.96
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培養条件の違いがウシ初期胚および胚盤胞期胚のヒストンアセチル化状態に及ぼす影響. Reviewed
後藤大也、岩田尚孝、橋本周、森本義晴、竹尾駿、門司恭典、桑山岳人
日本胚移植学雑誌 34 ( 3 ) 101 - 107 2012.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
胚の気相条件がヒストンのアセチル化状態に影響することを示した
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Differential attachment of bovine Y choromosome-beraing sperm to the zona pellucida. Reviewed
Y SAkaguchi, Y Sejiyama, S Kato, H Iwata, Y Monji, T Kuwayama.
Journal of mammalian ova reserch 29 ( 1 ) 55 - 59 2012.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
受精において透明帯に対する精子の接着性には、X染色体とY染色体を有する精子の間に差があることを牛の卵子と精子を用いて示した。
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Effect of maternal reproductive aging on ATP content and rate of in vitro fertilization of Japanese black cow oocytes Reviewed
Japanese Journal of Embryo Transfer 33 ( 3 ) 91 - 96 2011.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
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Effect of bovine age on the proliferative activity, global DNA methylation, relative telomere length and telomerase activity of granulosa cells Reviewed
H Goto, H Iwata, S Takeo, K Nisinosono, S Murakami, Yi Monji, T Kuwayama
Zygote 27 1 - 9 2011.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
牛では加齢に伴い顆粒層細胞の低メチル化、テロメアの短縮、テロメアーゼ活性の低下および増殖活性の低下が起こる。
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Trichostatin A-treated eight-cell bovine embryos had increased histone acetylation and gene expression, with increased cell numbers at the blastocyst stage Reviewed
Shiratsuki S ,Iwataa H*, Kimura K, Kuge T, Monji Y, Kuwayama T
Theriogenology 75 841 - 848 2011.02
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
トリコスタチンＡでウシ胚をゲノム活性化時期に処理することでヒストンのアセチル化が亢進しOct3/4 Cadherinを含む遺伝子発現が亢進し、ICMが多い胚が作成できる
DOI： 10.1016
Other Link： http://www.ncbi.nlm.nih.gov/pubmed/21144563
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The effect of maternal age on mitochondrial DNA copy number, ATP content and in vitro fertilization outcome of bovine oocytes Reviewed
Iwata H, Goto H, Tanaka H, Sakaguchi Y, Kimura K, Kuwayama T and Monji Y
Reproduction, Fertility and Development 23 424 - 432 2011.02
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
老齢のウシ由来の卵子のミトコンドリアが減少すること、異常受精が増えること ATPの生産が異常に更新することを示した
DOI： 1031-3613/11/030424
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Effect of N-Acetyl-D-Glucosamine on Bovine Sperm-Oocyte Interactions Reviewed
Yosuke Sakaguchi, Ryosuke Uzuhashi, Hisataka Iwata, Yasunori Monji and Takehito Kuwayama
Journal of Mannmalian Oova Reserch 27 ( 3 ) 130 - 135 2010.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
卵子の透明帯に接着する精子の数はを対象に、卵子の成熟に伴い透明帯の質が変化することを示した
Other Link： http://www.jstage.jst.go.jp/browse/jmor/27/3/_contents/-char/ja/
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Follicle growth and oocyte developmental competence in cows with liver damage. Reviewed
Iwata H, Tanaka H, Kanke T, Sakaguchi Y, Shibano K, Kuwayama T, Monji Y.
Reproduction in Domestic Animals 45 ( 5 ) 888 - 895 2010.10
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
肝臓に疾病を有する牛の血液、卵胞液の組成の異常を示し、卵巣内の卵子の発育と胞状卵胞内の卵子の受精率、発生率に異常が起こることを示した
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Study of in vitro culture conditions for bovine oocytes from early antral follicles Reviewed
Kanke T, Iwata H, Ohota M, Monji M, Kuwayama T
Japanease journal of embryo transfer 32 ( 3 ) 103 - 112 2010.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
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Effect of Maternal Age on the Developmental Competence and Progression of Nuclear Maturation in Bovine Oocytes
Yamamoto T, Iwata H, Goto H, Shiratuki S, Tanaka H, Monji Y, Kuwayama T
Moleciular Reproduction and Development 2010.06
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
老齢のウシ由来の卵子ではギャップ結合の喪失が早く、核成熟が早く進み、異常受精が多い。またＭＰＦの活性の動態も早いがサイクリンＢやＣＤＫ1およびMAD2の発現量には差が見られなかった
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ブタ着床前期胚の体外培養
佐々木康人・岩田尚孝・門司恭典・桑山岳人
日本胚移植学雑誌 2009.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
ブタの受精後１２日目の胚の体外培養には血清だけでなくBSAとの組み合わせやアミノ酸の添加が効果的であることを示した。
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Effect of N-Acetyl-D-Glucosamine on Bovine Sperm-Oocyte Interactions
Y Sakaguchi, H Iwata, T Kuwayama, Y Monji
Journal of Reproduction and Development 2009.09
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
ウシ精子と卵子の共培養条件にNアセチルグルコサミンを添加すると接着精子が減るのに多精子が増える。この理由は受精後の透明帯硬化を阻害することによることを示した
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凍結希釈液へのイクラ由来の脂質の添加が豚精子の性状に及ぼす影響
斉藤孝太・水野宏亮・岩田尚孝・桑山岳人・門司恭典
日本胚移植学雑誌 2009.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
イクラ由来の脂質成分は豚の精液に冷却耐性を与えることが示された
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Effect of carbohydrates on the ability of bull sperm to bind to bovine oviduct epithlial cells.
Kon Y, Iwata H, Shiono H, Matubara K, Kurita A, Y Sakaguchi, Kuwayama T, Monji Y.
Reproduction in Domestic Animals 2009.06
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
卵管上皮への結合性をＮアセチルＤグルコサミンによって昂進させることが出来るが、この接着は通常の接着とことなり精子の生存時間の延長に貢献しないことを示した。
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経皮吸収エストラジオール利用が黒毛和種経産牛の過剰排卵成績に及ぼす効果
坂上信忠、秋山清、中澤慶紀、橋村慎二、濱野岳人、久下壮、岩田尚孝、門司恭典
繁殖技術研究会誌 30 ( 3 ) 2008.09
Language：Japanese Publishing type：Research paper (scientific journal)
CIDRを用いた過剰排卵処理時にETSを併用することで採卵成績が向上することを示した。
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Effects of follicular fluids on the growth of porcine preantral follicle and oocyte
T Metoki, H Iwata*, M Itoh, M Kasai, A Takajyo, A Suzuki, T Kuwayama, Y Monji
Zygote 2008.08
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
卵胞培養に大卵胞由来の卵胞液を添加することで従来の血清を用いた培養方法よりも効率よく卵胞を発育させることが出来ることを示した。
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Effect of follicular fluid collected from various diameter follicles on the progression of nuclear maturation and developmental competence of pig oocytes.
Ito, M. Iwata H, Kitagawa, Y Kon, T. Kuwayama, Y. Monji
Animal Reproduction Science 2008.07
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
卵胞培養に大卵胞由来の卵胞液を添加することで従来の血清を用いた培養方法よりも効率よく卵胞を発育させることが出来ることを示した。
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ブタにおける精子処理および供培養時間が透明帯接着精子のY染色体を有する精子比率に及ぼす影響
塩野 紘・岩田尚孝・斉藤孝太・浦川真明・栗田敦史・今 義拓・佐々木康仁・桑山岳人・門司恭典
日本胚移植学雑誌 2008.05
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
ブタの新線、凍結精子を用いて透明帯への接着性がY染色体を有した精子において高く、この優位性が前培養処理によって低下することを示した
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ブタにおける精子処理および供培養時間が透明帯接着精子のY染色体を有する精子比率に及ぼす影響
塩野 紘、岩田尚孝、斉藤孝太、浦川真明、栗田敦史、今 義拓、佐々木康仁、桑山岳人、門司恭典
繁殖技術研究会誌 2008.05
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
ブタの新鮮、凍結精子を用いて透明帯への接着性がY染色体を有した精子において高く、この優位性が前培養処理によって低下することを示した。
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形態から卵胞委縮と推定されたウシの卵胞発育状態
中村光子、栗田敦史、増田恵、新井信元、岩田尚孝、桑山岳人、門司恭典
繁殖技術研究会誌 2008.05
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
早期閉経状態にあるウシ卵巣中の卵胞発育動態について観察し、卵胞発育が著しく低下していることを示した
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Effects of modification of in vitro fertilization techniques on the sex ratio of the resultant bovine embryos. Ani Reprod Sci 2007 accept Reviewed
Iwata H, Shiono H, Kon Y, Matubara K, Kimura K, Kuwayama T, Monji Y
Animal Reproduction Science 105 ( 3-4 ) 234 - 244 2008.03
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
体外受精時の精子と卵子の受精時間が短いと得られる胚にオスが多くなること示しこれが、精子の受精能獲得のタイミングの差に起因していることを示した。
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The effect of glucosamine concentration on the development and sex ratio of bovine embryos.
Kimura K, Iwata H, Thompson JG.Ｘ
Animal Reproduction Science 2008.02
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
染色体上の遺伝子にその代謝がコードされているグルコサミンを添加した培養条件では発生してくる胚の性別がオスに傾く
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メチルアセトアミドを耐凍剤に用いたニワトリ凍結精液による人工授精後の授精率の推移
伊藤 愛弓、 園田 将晴、岩田尚孝、門司 恭典、桑山 岳人
関東畜産学会誌 2007.10
Language：Japanese Publishing type：Research paper (scientific journal)
採血をおこなうとニワトリのストレスレベルが急速に向上し血中のコルチコステロン濃度が上昇することが示された。
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雄ニホンウズラの採血に対するコルチコステロン分泌反応
北園 大志、伊藤 哲、金子 佳恵、岩田 尚孝、門司 恭典、桑山 岳人
関東畜産学会 2007.10
Language：Japanese Publishing type：Research paper (scientific journal)
メチルアセトアミドを用いた凍結精液では受精後9日後までに急速に受胎率が低下することや、融解後の運動率の低下が著しいことを示した。、
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Successful superovulation of cattle by a single administration of FSH in aluminum hydroxide gel
Kimura K, Hirako M, Iwata H, Aoki M, Kawaguchi M, Seki M.
Theriogenology 2007.09
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
アルミニウムゲルを用いたホルモン処理がウシの過排卵にも有効であること示した。
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Responsiveness of Rabbits to Superovulation Treatment by Single Injection of Follicle Stimulating Hormone mixed with Aluminum Hydroxide-gel
Shu HASHIMOTO^1) , Koji KIMURA^2) , Takashi KURAMOCHI^1) , Kazuki
Molecular Reproduction and Development 2007.07
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
ウサギに対してアルミニウムゲルをアジュバンドに用いたホルモン処理が有効であることを示した。
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牛体外受精卵の性別の偏りについて
岩田 尚孝
日本胚移植学雑誌 2007.05
Language：Japanese Publishing type：Research paper (scientific journal)
胚の性判別や性操作の現状について解説した
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The effect of glucose on the progression of the nuclear maturation of pig oocytes
Sato H a, Iwata H a, Hayashi T a, Kimura K b, Kuwayama T a and Monji Y
Animal Reproduction Science 2007.04
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
グルコースがブタ卵子の体外成熟に解糖系だけでなくペントースリン酸回路を使って核成熟を促進することを示した
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Effect of Modification of Ovary Preservation Solution by Adding Glucose on the Maturation and Development of Pig Oocytes after Prolonged Storage.
Sakamoto A, Iwata H, Sato H, Hayashi T, Kuwayama T, Monji Y
Journal of Reproduction and Development 2006.11
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
卵巣の保存液にグルコースを添加することで長期的な保存による卵子の活力低下を防げることを示した
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Induction of estrus by prostaglandin F2a administration in the vaginal vestijble of miniature pigs
T. Kuge, H Iwata, T Kadokawa, I Domeki, Y Shioya, T Kuwayama, Y Monji
Journal of Reproduction and Development 2006.04
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
膣前庭粘膜へのPGF2の添加が発情の同期化に有効であることhcGの併用投与が同期化に及ぼす影響について示した
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産卵、抱卵、育雛中の岐阜地鶏における血漿コルチコステロン濃度
飯田真希 山本茜、岩田尚孝、桑山 岳人、門司 恭典
関東畜産学会報 2006.04
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal)
岐阜地鶏の抱卵が鳥に与えるストレスについて検討した
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Inhibitory effect of Ca-ionophore pretreatment of porcine oocytes on polyspermic fertilization
T Hayashi, H Sato, H Iwata, T Kuwayama, and Y monji
Zygote 2006.02
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
豚卵子を成熟後カルシウイオノフォア処理することで多精子受精を防げることを示した
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The localization of LAP2beta during pronuclear formation in bovine oocytes after fertilization or activation.
Isaji M, Iwata H, Harayama H, Miyake M
Zygote 2006.02
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
体細胞でのヒストンH3のリン酸化のパターンが卵子では体細胞のそれと異なることを示した またシクロヘキシミドはLAP2の配列を抑制することを示した
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Comparison between the characteristics of follicular fluid and the developmental competence of bovine oocytes
H Iwata*,J Inoue, K Kimura, T Kuge, T Kuwayama, Y Monji
Animal Reproduction Science 2006.01
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
卵巣内の卵子の発生能力や受精能力と卵胞液中の生理性状値を比較し黄疸指数や尿素態窒素含量と大きな相関があることを示した
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Modification of ovary stock solution with magnesium and raffinose improved the developmental competence of oocytes after long preservation
H Iwata1, T Hayashi, H Sato, K Kimura, T Kuwayama, and Y Monji
Zygote 2005.11
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
卵巣の保存方法について 保存液へのラフィノースやマグネシウムの添加が保存後の牛卵巣卵子の発生能力の保持に有効であることを示した。
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ミニチュアブタの黄体開花期におけるプロスタグランジンF2α連続投与が血中性ステロイドホルモン濃度及び膣内生理諸性状に及ぼす影響
久下壮、岩田尚孝、桑山岳人、門司恭典
東京農業大学農学集報 2005.09
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (bulletin of university, research institution)
ミニチュアブタの膣前庭へのPGF2α投与によって発情の同期化が可能であることを示した
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Effects of follicle size and electrolytes and glucose in maturation medium on nuclear maturation and developmental competence of bovine oocytes
Iwata H, Hashimoto S, Ohota M, Kimura K, Shibano K and Miyake M
Reproduction 127:159-164 2004.02
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
大卵胞の卵子は小卵胞の卵子に比べて発生能力が高い。そこでそれぞれの卵胞液の組成を測定するとNa K Glucoseにおいて差があった。また通常の培地と比較しMgの濃度が高かった。また大卵胞卵子は成熟速度が速く、発生能が高い事、小卵胞卵子の成熟において高いMgやグルコース濃度が早い成熟と高い発生能の獲得を支持することを示した
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The localization of LAP2(beta) in bovine oocytes after in vitro activation and fertilization
M Isaji, H Iwata, H Harayama, M Miyake.
Zygote 12：81-93 2004.01
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
LAP2、ラミンA/CとBの局在を単為発生胚と体外受精胚を用いて調べた DNAの合成時期とLAP2の発現時期は重なっているほか、体外受精卵においても単為発生胚においても核膜に存在した。またラミンA/Cは単為発生の内細胞隗には存在したが体外受精胚には存在しなかった
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Effect of growth hormone at various periods of development on developmental competence of bovine embryo and its interaction with glucose
Iwata H, Ohota M, Hashimoto S, Kimura K, Isaji M, Miyake M.
Journal of Reproduction and Development 49 6巻 （2003) 493-499 2003.12
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
胚の体外培養を改善するため、FCSやBSAの血清蛋白を用いた培養条件でGrowth Hormon の添加とグルコースの濃度の胚発生に及ぼす影響について発生ステージごとに検討した。FCS添加条件下でGHは桑実胚以降において発生を促進しすること、またGHの効果は培養条件中のグルコースの濃度と相互効果が見られること、桑実胚以降のグルコース濃度の上昇は胚発生促進には効果がないこと等を示した。
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Free Oxygen radicals are generated at the time of aspiration of oocytes from ovaries that have been stored for a long time
Iwata H, Ohota M, Hashimoto S, Yoshiharu N.
Zygote 11；1-5 2003.02
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
卵巣の保存時間の延長に伴い卵巣卵子の活力は低下する。高温（30度以上）で保存した卵巣から卵子を回収する時、抗酸化剤を併用することで回収時の卵子の活力低下が防げることを示し、長期間保存した卵巣から卵子を回収する際に虚血再還流時と同様に活性酸素が産生していること示した
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Developmental competence of bovine oocytes arrested at germinal vesicle stage by cycloheximide under air
Hashimoto S, Kimura K, Iwata H, Takakura R
Journal of Reproduction and Development 49; 61-66 2003.02
Language：English Publishing type：Research paper (scientific journal)
シクロヘキシミドで卵子を処理し、20度で24時間GV期で保持したところ、その後卵子の発生能力は対象区のそれと変わらず、卵子の輸送方法として有効であることを示した
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Localization of Lap2B in oocytes and embryos during early development in cattle
Isaji M,Iwata H, Ohota M, Harayama H and Miyake M.
Theriogenology 第59巻 474 (abstr) 2003.01
Language：English Publishing type：Research paper (scientific journal)
LAP2βはラミンの裏打ち蛋白であるがその機能については不明な点が多い。ウシ胚においてこのLAP2βの局在と発現について観察した。LAP2βの発現や局在は体外受精卵や単為発生胚で差がなく発生が進むにつれて前核期よりも発現が強くなることを示した。
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Glucose-6-phosphate dehydrogenase activity of IVM/IVF/IVC bovine embryos is related to the sex and developmental competence under suboptimal gas condition
Iwata H, Kimura K, Hashimoto S, Oota M, Tominaga T and Minami N.
Journal of Reproduction and Development 5 447 - 453 2002.10
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
Brilliant Cresyl Blueを用いて発生各ステージの体外受精卵の還元力の違いと性比との関係について検討した。その結果桑実胚において還元力の差が雄と雌との間で異なること、8細胞期や胚盤胞期では差がないことを示した。このことより桑実期胚ではX染色体上にあるG6PDの活性に差があり雌雄判別に利用できることやX染色体の不活化が胚盤胞期胚以降で起こっていることを示唆した。
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Postnatal weights of calves derived from IVM/IVF embryos developed under various oxygen concentrations.
Iwata H, Minami N, Imai H
Reproduction Fertility and Development 12 391 - 396 2000.12
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
体外受精卵に由来する仔牛体重の大型化は広く知られている。同一の精液、同系統の母牛を用いた均一な牛群を用いて実験を行い、高酸素の気相条件dでは、低酸素条件に比べて体外受精卵由来産子の体重が大型化すること、またこの産子体重の大型化は妊娠期間の延長は伴わないことを示した。
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Allopurinol, an inhibitor of xanthine oxidase, improves the development of IVM/IVF bovine embryos (>4 cell) in vitro under certain culture condition
Iwata H, Akamatsu S, Minami N
Theriogenology 51 613 - 622 1999.04
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
ウシ胚の体外培養において胚発生に悪影響を与える活性酸素の一部はキサンチンオキシダーゼ系に由来することを示した。またこの活性酸素の産生は培養条件中のグルコースや高酸素の気相条件によって活性化することを示した。
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Comparison of weight of calves derived from embryos developed in vivo or in vitro with different culture media.
Iwata H, Akamatsu S and Minami N
Journal of Reproduction and Development 44 33 - 38 1998.10
Authorship：Lead author Language：Japanese Publishing type：Research paper (scientific journal)
ウシの産子体重が使用する系統（但馬、島根系）によって大きく異なることを示し、同一系統の卵巣、同一の精液を用い体外受精卵及び体内受精卵の産子体重と妊娠期間について検討した。そして体外受精卵由来産子の体重が妊娠期間の延長とかかわらず延長することを示した。また発生培養液（TCM-199とｍ-SOF)によって産子体重の増加の度合いに差があることを示した
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Effects of antioxidants on the development of bovine IVM/IVF embryos in various concentration of glucose.
Iwata H, Akamatsu S, Minami N, Yamada M
Theriogenology 50 365 - 375 1998.02
Authorship：Lead author Language：English Publishing type：Research paper (scientific journal)
ウシ胚の体外培養において、培養気相条件中の酸素濃度が高い場合や培地中のグルコースの濃度が高い場合に活性酸素が産生し、胚に悪影響を及ぼしていることを示した
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共培養を用いた体外成熟・体外受精由来牛受精卵の発生に及ぼすタウリンの影響
岩田尚孝・岩藤勝彦・内海恭三
日本胚移植研究会誌 1995.08
Authorship：Lead author Language：Japanese Publishing type：Research paper (scientific journal)
ウシ体外受精卵の発生培養液へのタウリンの影響を、共培養、コンディション培地を用いて検討した。タウリンの添加により共培養を用いた培養系で安定的な胚発生成績を得ることができるようになり、新鮮卵や凍結卵の移植成績も改善された