Journal of Reproduction and Development最新文献

{"title":"Large cyclic AMP-dependent increases in tyrosine-phosphorylated proteins are not absolutely necessary for the induction of full-type hyperactivation and penetration into eggs in ejaculated boar spermatozoa in vitro.","authors":"Ami Ueshiba, Riko Nakao, Riho Morikawa-Tomita, Hirohisa Kyogoku, Hiroshi Harayama","doi":"10.1262/jrd.2025-092","DOIUrl":"10.1262/jrd.2025-092","url":null,"abstract":"<p><p>Although it is well-known that sperm tyrosine-phosphorylated proteins increase in a cyclic AMP (cAMP)-dependent manner during capacitation, it remains unclear whether or not this reaction is involved directly in sperm fertilization-related events. This study aimed to examine the effects of pharmacological suppression of cAMP-dependent increases in tyrosine-phosphorylated proteins on the induction of full-type hyperactivation and penetration into eggs in boar spermatozoa. Ejaculated spermatozoa were treated with a cAMP analog (cBiMPS) and subsequently used for Western blotting, flagellar beating assays, and egg penetration assays. Marked increases in tyrosine-phosphorylated proteins and full-type hyperactivation were observed in spermatozoa treated with cBiMPS in the presence of CaCl₂. These spermatozoa effectively penetrated eggs. A spleen tyrosine kinase (SYK) inhibitor OXSI2 suppressed large cBiMPS-dependent increases in sperm tyrosine-phosphorylated proteins, in contrast to the ineffectiveness of SU6656 (a SRC family kinase inhibitor) and dasatinib (a c-ABL inhibitor). However, this suppression was not associated with changes in the state of full-type hyperactivation or the outcomes of egg penetration assays conducted in insemination medium without OXSI2. Conversely, adding OXSI2 to both the cBiMPS treatment medium and the sperm insemination medium suppressed increases in sperm tyrosine-phosphorylated proteins during insemination and significantly decreased the average number of spermatozoa that penetrated eggs. These results suggest that high SYK activation and large cAMP-dependent increases in tyrosine-phosphorylated proteins are not absolutely necessary for inducing full-type hyperactivation and penetration into eggs in boar spermatozoa in vitro, but may influence sperm functions associated with the outcomes of egg penetration assays.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":"55-64"},"PeriodicalIF":2.2,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13078923/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146202041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional roles of NR4A transcription factors in GnRH regulation of gonadotropin gene expression and secretion in rat primary pituitary cells.","authors":"Ryota Terashima, Yuta Tomiyama, Shiro Kurusu, Mitsumori Kawaminami","doi":"10.1262/jrd.2025-100","DOIUrl":"10.1262/jrd.2025-100","url":null,"abstract":"<p><p>Gonadotropin-releasing hormone (GnRH) tightly regulates the synthesis and the release of gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), while the intracellular molecular mechanisms following GnRH signal for the regulation of transcription remain incompletely understood. In this study, we used primary culture of rat anterior pituitary cells to investigate the role of NR4A transcription factors (NR4A1, NR4A2, and NR4A3) in GnRH regulation of gonadotropin secretion. GnRH agonist stimulation rapidly and transiently increased Nr4a1, Nr4a2, and Nr4a3 expression within one hour, accompanied by a time-dependent increase in Fshb mRNA levels and the secretion of both FSH and LH. The knockdown of each Nr4a gene using siRNA significantly reduced Fshb expression under GnRH stimulation. Nr4a1 knockdown caused the most pronounced decrease in FSH secretion. Although Lhb and Cga mRNA levels were largely unaffected, LH secretion was consistently reduced following NR4A knockdown. These findings suggest that NR4A transcription factors act downstream of GnRH signaling to promote Fshb transcription and facilitate gonadotropin secretion, thereby modulating GnRH-dependent control of FSH and LH secretion.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":"48-54"},"PeriodicalIF":2.2,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13078925/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145985102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Controlled internal drug release (CIDR) removal before luteolysis prolongs estrus interval and increases silent heat risk in Japanese Black cows.","authors":"Takuo Hojo, Kei Horihata, Naoki Takenouchi","doi":"10.1262/jrd.2025-076","DOIUrl":"10.1262/jrd.2025-076","url":null,"abstract":"<p><p>This study aimed to improve estrus management in Japanese Black cows by investigating the effects of controlled internal drug release (CIDR) treatment on luteal function including luteolysis and estrous characteristics. Cows with normal estrous cycles were divided into three groups, and CIDR treatment was initiated on day 5 (Group 1), 10 (Group 2), or 16 (Group 3) of the estrous cycle and continued for 12 days. Plasma progesterone (P₄) levels, luteal diameter, and luteal blood flow (LBF) were monitored during treatment, and correlations among these parameters were analyzed. Additionally, the interval from CIDR removal to estrus, and the incidence of standing (ST) behavior during estrus were evaluated. All luteal parameters changed in a cycle-dependent manner, irrespective of CIDR treatment. Significant correlations were observed between P₄ levels and both luteal diameter and LBF. The interval from treatment withdrawal to estrus was significantly longer in Group 1 (4.6 ± 2.2 days) than in Groups 2 (2.1 ± 0.4 days) and 3 (2.2 ± 0.7 days). Silent heat, defined as estrus without ST behavior, occurred in 27.3% of cows in group 1 and 10.0% in Group 2, but not in Group 3. These findings suggest that initiating CIDR treatment in the early luteal stage and removing it just before or during luteolysis, causes variability in estrus timing after treatment and increases the risk of silent heat. When implementing estrus synchronization using CIDR in the field, both the timing of treatment initiation and removal should be carefully considered.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":"65-72"},"PeriodicalIF":2.2,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13078928/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146220204","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Niacin improves developmental ability by increasing glutathione levels and promoting male pronuclear formation in porcine oocytes grown in vitro.","authors":"Takayuki Yamochi, Shu Hashimoto, Yoshiharu Morimoto","doi":"10.1262/jrd.2025-140","DOIUrl":"10.1262/jrd.2025-140","url":null,"abstract":"<p><p>We examined the effect of niacin, a water-soluble vitamin essential for the metabolism of carbohydrates, lipids, and other substrates, on the developmental potential of oocytes grown in vitro, as well as the number and activity of mitochondria and reduced glutathione (GSH) content in oocytes. Porcine oocytes were cultured in medium containing 0-10 mM niacin for 12 days. Subsequently, in vitro maturation was performed, followed by in vitro fertilization using intracytoplasmic sperm injection, and the ability to develop into blastocysts was examined. Niacin treatment (10 mM) increased the mitochondrial, GSH, and glutamate-cysteine ligase catalytic subunit contents in porcine oocytes grown in vitro. Furthermore, niacin treatment enhanced male pronuclear formation and blastocyst development rates in porcine oocytes grown in vitro. Niacin increased the mitochondrial content and GSH levels in porcine oocytes and enhanced the ability of oocytes to form male pronuclei, which in turn promoted embryonic development.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":"73-79"},"PeriodicalIF":2.2,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13078926/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147276545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of paternal resveratrol intake on mitochondria and telomere length in mouse embryos and offspring.","authors":"Noko Teramoto, Nanako Kobayashi, Kokoa Mitsui, Asuka Kamio, Komei Shirasuna, Hisataka Iwata","doi":"10.1262/jrd.2026-038","DOIUrl":"https://doi.org/10.1262/jrd.2026-038","url":null,"abstract":"<p><p>Advanced paternal age affects embryonic development and offspring phenotypes. We previously demonstrated that resveratrol prevents age-associated declines in the mitochondrial DNA copy number (mt-cn) and telomere length (TL) in embryos. The present study was performed to investigate the effects of paternal resveratrol intake on the mt-cn and TL in pups and embryos produced either via in vitro fertilization (IVF) or IVF using sperm preincubated in epididymal fluid (EF). C57BL/6N male mice were administered drinking water containing either the vehicle (ethanol, 1/2500) or 0.1 mM resveratrol. When these males were mated with young ICR females, paternal resveratrol treatment did not affect the mt-cn or TL in offspring derived from young fathers (16-25 weeks of age). In contrast, in offspring of aged fathers (41-51 weeks of age), the mt-cn and TL in heart tissue were altered in a sex-dependent manner. Moreover, continuous resveratrol administration to males until an advanced paternal age resulted in sperm TL elongation. However, resveratrol treatment did not affect the TL in embryos but significantly increased the mt-cn and reduced the lipid content in blastocysts produced via IVF using oocytes from young females (4 weeks of age). RNA- sequencing revealed that resveratrol treatment affected metabolic pathways, including lipid metabolism. The preincubation of sperm from young untreated males (11 weeks of age) in EF derived from resveratrol-treated males did not affect the mt-cn or TL in blastocysts but reduced the lipid content. In conclusion, paternal resveratrol intake modulates embryonic and offspring phenotypes, potentially through alterations in sperm TL, sperm epigenetic modifications, and bioactive components in EF. Resveratrol intake may exert long-term effects on offspring.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147623345","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sex determination and genetic screening of equine embryos with whole genome amplification and real-time PCR.","authors":"Gus R McFarlane, Elizabeth Backhouse, Agustin Ruiz, Brendon A O'Rourke, Jenin V Cortez, Christopher G Grupen","doi":"10.1262/jrd.2026-014","DOIUrl":"https://doi.org/10.1262/jrd.2026-014","url":null,"abstract":"<p><p>Reliable pre-implantation sex determination and genetic screening enables informed embryo transfer decisions in equine breeding while avoiding later interventions. We developed a streamlined workflow that couples rapid whole genome amplification (WGA) with a multiplex real-time PCR targeting ETSTY5 as a Y-specific marker and UBC as an autosomal control. On purified equine DNA, sex was correctly assigned down to 10 pg gDNA and to a single fibroblast cell. Direct testing of embryo biopsies without WGA yielded inconsistent results, whereas introducing a short WGA step produced tight allelic-discrimination clusters and 100% diagnostic calls, including in cloned embryos of known sex. The same WGA product supported targeted genotyping for inherited disease screening of hyperkalemic periodic paralysis (HYPP) and hereditary equine regional dermal asthenia (HERDA) alleles. This WGA plus real-time PCR pipeline supports robust and practical embryo sexing and targeted pre-implantation genetic diagnostics within in vitro produced (IVP) equine embryo and embryo transfer workflows.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147499179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ovarian and follicular culture enables the production of fertile oocytes in C57BL/6 mice.","authors":"Kyota Yoshida, Minami Fukuzawa, Yayoi Obata","doi":"10.1262/jrd.2026-008","DOIUrl":"https://doi.org/10.1262/jrd.2026-008","url":null,"abstract":"<p><p>In vitro experiments have become a powerful alternative to in vivo approaches for analyzing gene functions and molecular mechanisms. However, successful in vitro oogenesis using the C57BL/6 strain-widely used as a standard mouse model in mammalian studies-has not yet been reported. In this study, we established a 30-day culture system combining ovarian and follicular culture, and successfully produced fully competent oocytes from neonatal C57BL/6N ovaries, which contain only non-growing oocytes prior to primordial follicle formation. A key contributing factor to this success was the supplementation of dibutyryl cyclic AMP, a membrane-permeable analogue of endogenous cyclic AMP, which promoted follicular growth. This culture system provides a valuable tool for investigating the mechanisms underlying oogenesis.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147463561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"N-myristoyltransferase 1, a key gene for protein N-myristoylation, is dispensable for fertility in male mice.","authors":"Yang Liu, Yvxia Zhang, Hailing Ding, Jie Li, Tingting Gao, Zongzhuang Wen, Yunshan Wang, Bin Wu","doi":"10.1262/jrd.2025-064","DOIUrl":"https://doi.org/10.1262/jrd.2025-064","url":null,"abstract":"<p><p>N-Myristoyltransferase 1 (NMT1), the predominant enzyme catalyzing myristoylation of proteins, is involved in various biological processes, including early embryonic development, immune responses, apoptosis, cellular homeostasis, tumorigenesis, and infection, with therapeutic potential in viral and parasitic infections as well as cancer. Despite the critical functions of NMT1, there have been no reports to date regarding its role in reproduction, especially in spermatogenesis. To investigate the function of NMT1 in this context, we utilized CRISPR/Cas9 technology to create a germ cell-specific Nmt1 knockout mice model for the first time. Surprisingly, male mice lacking NMT1 maintained fertility, exhibiting normal testicular structure and sperm morphology, with no significant differences in spermatogenic tubule structure or germ cell distribution compared to wild-type mice. Additionally, the Nmt1^f/f; Stra8-Cre male mice showed no notable defects in meiosis. These findings suggest that NMT1 is not critical for spermatogenesis or male fertility in mice. However, further studies have shown that the compensatory role of NMT2 may play an unexpected role in maintaining myristoylation levels, which provides a new perspective for understanding the role of myristoylation in spermatogenesis.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147377903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interactions among progesterone, estradiol, and γ-aminobutyric acid in rat spermatozoal hyperactivation.","authors":"Miki Muroi, Shiho Fukuda, Miki Takahashi, Tatsuya Suzuki, Masakatsu Fujinoki","doi":"10.1262/jrd.2025-106","DOIUrl":"https://doi.org/10.1262/jrd.2025-106","url":null,"abstract":"<p><p>Our previous study in hamsters indicated that progesterone (P) induces enhancement of spermatozoal hyperactivation, while 17β-estradiol (Eβ) and γ-aminobutyric acid (GABA) suppress this enhancement. Since P has also been shown to enhance hyperactivation of rat spermatozoa in another study conducted by our team, we examined the effects of Eβ and GABA on the P-enhanced hyperactivation of rat spermatozoa in the present study. Our results showed that Eβ suppressed P-enhanced hyperactivation in a dose-dependent manner through an estrogen receptor. In contrast, GABA did not affect P-enhanced hyperactivation. Instead, 5-500 fM GABA significantly enhanced spermatozoal hyperactivation, and this GABA-induced enhancement of hyperactivation was associated with a GABA_A receptor. In conclusion, P-enhanced hyperactivation of rat spermatozoa was suppressed by Eβ although it was not suppressed by GABA. Moreover, low concentrations of GABA enhanced rat spermatozoal hyperactivation.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147377967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of thiol compounds on DNA-damage detection and in vitro fertilization outcomes in bovine sperm.","authors":"Kumiko Takeda, Kazuko Ogata, Kazutsugu Matsukawa, Hiroaki Yokokawa, Yutaka Fukumoto, Kenji Morimasa, Takemasa Hidaka, Takuya Nogi, Kenichi Watabe, Kumi Iijima, Ryoichi Nakamura, Masaki Ozawa, Masahiro Kaneda","doi":"10.1262/jrd.2025-138","DOIUrl":"https://doi.org/10.1262/jrd.2025-138","url":null,"abstract":"<p><p>Reduced glutathione (GSH), a thiol compound, plays an important role in protecting sperm from excessive levels of reactive oxygen species. Our previous study demonstrated that GSH supplementation was associated with improved in vitro fertilization (IVF) outcomes. However, GSH supplementation also increased the proportion of DNA-damaged sperm detected in the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, indicating involvement of its thiol components. In this study, we investigated the effects of several thiol compounds, including l-cysteine (Cys), N-acetyl-l-cysteine (NAC), and dithiothreitol (DTT), on bovine sperm DNA integrity. The addition of Cys or NAC (0.5, 1, or 5 mM) to the medium caused an increase in DNA damage, which was observed either throughout the sperm head or specifically in the post-acrosome (PA) region. DTT treatment increased DNA damage in both the PA and acrosomal regions, and it induced progressive sperm head lysis from the acrosome to the entire head. These treatments caused sperm DNA-damage rates that exceeded the normal range, suggesting that thiol groups enhance the detection of DNA damage. The effects of these thiols on IVF outcomes were further examined. Short-term exposure to 1 mM Cys before insemination improved cleavage rates, whereas prolonged exposure at concentrations ≥ 1 mM during insemination reduced cleavage rates. NAC supplementation had no significant effects. A mild negative correlation was observed between TUNEL-positive sperm rates and both cleavage and blastocyst formation rates following short-term treatment with 0.5 mM Cys. These findings suggest that thiol supplementation, when optimally dosed and timed, may enhance the sensitivity of DNA-damage detection rather than induce additional damage.</p>","PeriodicalId":16942,"journal":{"name":"Journal of Reproduction and Development","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2026-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147355454","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}