Papers - IWATA Hisataka
-
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
-
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.
-
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
-
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.
-
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
-
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
-
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.
-
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
-
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
-
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
-
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.
-
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.
-
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。
-
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.
-
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
-
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.
-
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.
-
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.
-
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.
-
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