The Journal of Reproduction and Development最新文献_第2页

Expression and localization of tight junction-related proteins in adult rat pituitary stem/progenitor cell niches 成年大鼠垂体干/祖细胞龛中紧密连接相关蛋白的表达和定位

The Journal of Reproduction and Development Pub Date : 2022-04-12 DOI: 10.1262/jrd.2021-150

S. Yoshida, Hideaki Yurino, Masaaki Kobayashi, Naoto Nishimura, K. Yano, K. Fujiwara, S. Hashimoto, T. Kato, Y. Kato

{"title":"Expression and localization of tight junction-related proteins in adult rat pituitary stem/progenitor cell niches","authors":"S. Yoshida, Hideaki Yurino, Masaaki Kobayashi, Naoto Nishimura, K. Yano, K. Fujiwara, S. Hashimoto, T. Kato, Y. Kato","doi":"10.1262/jrd.2021-150","DOIUrl":"https://doi.org/10.1262/jrd.2021-150","url":null,"abstract":"Pituitary endocrine cells are supplied by Sox2-expressing stem/progenitor cells in the anterior lobe of the adult pituitary gland. These SOX2-positive cells are maintained in two types of microenvironments (niches): the marginal cell layer (MCL)-niche and the parenchymal-niche. Recently, we isolated dense SOX2-positive cell clusters from the parenchymal-niche by taking advantage of their resistance to protease treatment as parenchymal stem/progenitor cell (PS)-clusters. In the present study, by analyzing these isolated PS-clusters, we attempted to identify novel structural characteristics of pituitary stem/progenitor cell niches. Quantitative real-time PCR showed that tight junction-related genes were distinctly expressed in the isolated PS-clusters. Immunocytostaining showed that the tight junction molecules, ZO-1 and occludin, were localized in the apical membrane facing the pseudo-follicle-like structure of the isolated PS-clusters regardless of the expression of S100β, which distinguishes the sub-population of SOX2-positive cells. Furthermore, immunohistochemistry of the pituitary glands of adult rats clearly demonstrated that ZO-1 and occludin were densely present in the parenchymal-niche encircling the pseudo-follicle, while they were observed in the apical membrane in the MCL-niche facing the residual lumen. Collectively, these tight junction-related proteins might be involved in the architecture and maintenance of the plasticity of pituitary stem/progenitor cell niches.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"67 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123624838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Insights into the NAD+ biosynthesis pathways involved during meiotic maturation and spindle formation in porcine oocytes 猪卵母细胞减数分裂成熟和纺锤体形成过程中NAD+生物合成途径的研究

The Journal of Reproduction and Development Pub Date : 2022-03-28 DOI: 10.1262/jrd.2021-130

C. Pollard, Ashleigh Younan, A. Swegen, Z. Gibb, C. Grupen

{"title":"Insights into the NAD+ biosynthesis pathways involved during meiotic maturation and spindle formation in porcine oocytes","authors":"C. Pollard, Ashleigh Younan, A. Swegen, Z. Gibb, C. Grupen","doi":"10.1262/jrd.2021-130","DOIUrl":"https://doi.org/10.1262/jrd.2021-130","url":null,"abstract":"Treatments that elevate NAD+ levels have been found to improve oocyte quality in mice, cattle, and pigs, suggesting that NAD+ is vital during oocyte maturation. This study aimed to examine the influence of different NAD+ biosynthetic pathways on oocyte quality by inhibiting key enzymes. Porcine oocytes from small antral follicles were matured for 44 h in a defined maturation system supplemented with 2-hydroxynicotinic acid [2-HNA, nicotinic acid phosphoribosyltransferase (NAPRT) inhibitor], FK866 [nicotinamide phosphoribosyltransferase (NAMPT) inhibitor], or gallotannin [nicotinamide mononucleotide adenylyltransferase (NMNAT) inhibitor] and their respective NAD+ pathway modulators (nicotinic acid, nicotinamide, and nicotinamide mononucleotide, respectively). Cumulus expansion was assessed after 22 h of maturation. At 44 h, maturation rates were determined and mature oocytes were fixed and stained to assess spindle formation. Each enzyme inhibitor reduced oocyte maturation rate and adversely affected spindle formation, indicating that NAD+ is required for meiotic spindle assembly. Furthermore, NAMPT and NMNAT inhibition reduced cumulus expansion, whereas NAPRT inhibition affected chromosomal segregation. Treating oocytes with gallotannin and nicotinamide mononucleotide together showed improvements in spindle width, while treating oocytes with 2-HNA and nicotinic acid combined showed an improvement in both spindle length and width. These results indicate that the salvage pathway plays a vital role in promoting oocyte meiotic progression, while the Preiss-Handler pathway is essential for spindle assembly.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"13 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122823816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Central somatostatin-somatostatin receptor 2 signaling mediates lactational suppression of luteinizing hormone release via the inhibition of glutamatergic interneurons during late lactation in rats 中央生长抑素-生长抑素受体2信号通过抑制大鼠泌乳后期谷氨酸能中间神经元介导促黄体生成素释放的泌乳抑制

The Journal of Reproduction and Development Pub Date : 2022-03-06 DOI: 10.1262/jrd.2022-009

A. Sugimoto, Hitomi Tsuchida, Mayuko Nagae, N. Inoue, Y. Uenoyama, H. Tsukamura

{"title":"Central somatostatin-somatostatin receptor 2 signaling mediates lactational suppression of luteinizing hormone release via the inhibition of glutamatergic interneurons during late lactation in rats","authors":"A. Sugimoto, Hitomi Tsuchida, Mayuko Nagae, N. Inoue, Y. Uenoyama, H. Tsukamura","doi":"10.1262/jrd.2022-009","DOIUrl":"https://doi.org/10.1262/jrd.2022-009","url":null,"abstract":"Reproductive function is suppressed during lactation owing to the suckling-induced suppression of the kisspeptin gene (Kiss1) expression in the arcuate nucleus (ARC) and subsequent suppression of luteinizing hormone (LH) release. Our previous study revealed that somatostatin (SST) neurons mediate suckling-induced suppression of LH release via SST receptor 2 (SSTR2) in ovariectomized lactating rats during early lactation. This study examined whether central SST-SSTR2 signaling mediates the inhibition of ARC Kiss1 expression and LH release in lactating rats during late lactation and whether the inhibition of glutamatergic neurons, stimulators of LH release, is involved in the suppression of LH release mediated by central SST-SSTR2 signaling in lactating rats. A central injection of the SSTR2 antagonist CYN154806 (CYN) significantly increased ARC Kiss1 expression in lactating rats on day 16 of lactation. Dual in situ hybridization revealed that few ARC Kiss1-positive cells co-expressed Sstr2, and some of the ARC Slc17a6 (a glutamatergic neuronal marker)-positive cells co-expressed Sstr2. Furthermore, almost all ARC Kiss1-positive cells co-expressed Grin1, a subunit of N-methyl-D-aspartate (NMDA) receptors. The numbers of Slc17a6/Sstr2 double-labeled and Slc17a6 single-labeled cells were significantly lower in lactating dams than in non-lactating rats whose pups had been removed after parturition. A central injection of an NMDA antagonist reversed the CYN-induced increase in LH release in lactating rats. Overall, these results suggest that central SST-SSTR2 signaling, at least partly, mediates the suppression of ARC Kiss1 expression and LH release by inhibiting ARC glutamatergic interneurons in lactating rats.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"331 5","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"120840138","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Involvement of Ca2+-ATPase in suppressing the appearance of bovine helically motile spermatozoa with intense force prior to cryopreservation 参与Ca2+- atp酶在抑制牛螺旋运动精子的外观与强力之前冷冻保存

The Journal of Reproduction and Development Pub Date : 2022-03-03 DOI: 10.1262/jrd.2021-143

Duritahala, M. Sakase, H. Harayama

{"title":"Involvement of Ca2+-ATPase in suppressing the appearance of bovine helically motile spermatozoa with intense force prior to cryopreservation","authors":"Duritahala, M. Sakase, H. Harayama","doi":"10.1262/jrd.2021-143","DOIUrl":"https://doi.org/10.1262/jrd.2021-143","url":null,"abstract":"In cattle, cryopreserved spermatozoa are generally used for artificial insemination (AI). Many of these specimens exhibit helical movement, although the molecular mechanisms underlying this phenomenon remain unclear. This study aimed to characterize helically motile spermatozoa, investigate the involvement of Ca2+-ATPase in suppressing the appearance of these spermatozoa prior to cryopreservation, and examine the potential of helical movement as an index of sperm quality. In the cryopreserved semen, approximately 50% of spermatozoa were helically motile, whereas approximately 25% were planarly motile. The helically motile samples swam significantly faster than those with planar movement, in both non-viscous medium and viscous medium containing polyvinylpyrrolidone. In contrast, in non-cryopreserved semen, planarly motile spermatozoa outnumbered those that were helically motile. Fluorescence microscopy with Fluo-3/AM and propidium iodide showed that flagellar [Ca2+]i was significantly higher in cryopreserved live spermatozoa than in non-cryopreserved live ones. The percentage of non-cryopreserved helically motile spermatozoa was approximately 25% after washing, and this increased significantly to approximately 50% after treatment with an inhibitor of sarcoplasmic reticulum Ca2+-ATPases (SERCAs), “thapsigargin.” Immunostaining showed the presence of SERCAs in sperm necks. Additionally, the percentages of cryopreserved helically motile spermatozoa showed large inter-bull differences and a significantly positive correlation with post-AI conception rates, indicating that helical movement has the potential to serve as a predictor of the fertilizing ability of these spermatozoa. These results suggest that SERCAs in the neck suppress the cytoplasmic Ca2+-dependent appearance of helically motile spermatozoa with intense force in semen prior to cryopreservation.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"12 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114289425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Effects of hypercaloric diet-induced hyperinsulinemia and hyperlipidemia on the ovarian follicular development in mice 高热量饮食诱导的高胰岛素血症和高脂血症对小鼠卵巢卵泡发育的影响

The Journal of Reproduction and Development Pub Date : 2022-03-02 DOI: 10.1262/jrd.2021-132

Qiao-li Zhang, Yan Wang, Jian-Sheng Liu, Yanxiao Du

{"title":"Effects of hypercaloric diet-induced hyperinsulinemia and hyperlipidemia on the ovarian follicular development in mice","authors":"Qiao-li Zhang, Yan Wang, Jian-Sheng Liu, Yanxiao Du","doi":"10.1262/jrd.2021-132","DOIUrl":"https://doi.org/10.1262/jrd.2021-132","url":null,"abstract":"Long-term hypercaloric diets may adversely affect the development of ovarian follicles. We investigated the effects of high sugar (HS), high fat low sugar (HFLS), and high fat normal sugar (HFNS) diets on the ovarian follicle development in mice fed with these diets as compared to those fed with normal diet (control) for 180 days. Body weight, gonadal fat, glucose, lipid, insulin, estrous cycle, sex hormones and ovarian tissues were examined, and metabolism-related protein expression in the ovaries was evaluated by immunoblotting. The mice fed with hypercaloric diets showed hyperinsulinemia and hyperlipidemia, and exhibited heavier body and gonadal fat weights, longer estrous cycles, and fewer preantral and antral follicles than mice fed with normal diet. The sex hormone levels in the blood were similar to those in controls, except for significantly elevated estradiol levels in the HS diet group. The AMPKα phosphorylation was reduced, while AKT phosphorylation and caspase-3 levels were increased in the ovarian tissues of mice in all three hypercaloric diet groups than those in control. Taken together, the results suggest hyperinsulinemia and hyperlipidemia as possible mechanisms that impair the development of ovarian follicles in response to long-term exposure to unhealthy hypercaloric diets.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"30 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-03-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134635927","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Leptin receptor expression and its change in association with the normalization of EGF profile after seminal plasma treatment in repeat breeder dairy cows 重复种奶牛精浆处理后瘦素受体表达及其变化与EGF水平正常化的关系

The Journal of Reproduction and Development Pub Date : 2022-02-28 DOI: 10.1262/jrd.2021-142

N. Ninpetch, Dagvajamts Badrakh, HAY MAR KYAW, Kohei Kawano, Y. Yanagawa, M. Nagano, S. Katagiri

{"title":"Leptin receptor expression and its change in association with the normalization of EGF profile after seminal plasma treatment in repeat breeder dairy cows","authors":"N. Ninpetch, Dagvajamts Badrakh, HAY MAR KYAW, Kohei Kawano, Y. Yanagawa, M. Nagano, S. Katagiri","doi":"10.1262/jrd.2021-142","DOIUrl":"https://doi.org/10.1262/jrd.2021-142","url":null,"abstract":"Factors associated with high milk production levels have been linked to alterations in the endometrial epidermal growth factor (EGF) profile, a cause of reduced fertility in dairy cows. Therefore, we examined the leptin system that connects nutritional status and reproduction in dairy cattle related to reduced fertility in repeat breeder cows. Plasma leptin concentrations were measured in 18 heifers, 20 high-yielding control cows, and 26 repeat breeder cows, showing an altered EGF profile. Then, all repeat breeder cows were infused with seminal plasma (SP) into the vagina at the next estrus to normalize the EGF profile, while heifers and control cows were infused with vehicle alone. All animals were examined for EGF profiles. Eighteen repeat breeder cows, nine heifers, and nine control cows were also determined for leptin receptor (Ob-R) expression levels in the estrous cycle before and after the infusion. SP normalized the EGF profile in 53.8% of the repeat breeder cows. Leptin concentrations were similar in all groups, regardless of the treatment results for the EGF profile. In contrast, Ob-R levels in repeat breeder and control cows were similar and higher than those in heifers before SP treatment. Ob-R in repeat breeders showing a normal EGF profile after treatment decreased to an intermediate level between heifers and control cows and may provide a clue to take measures against repeat breeding in dairy cows.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"10 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128647661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Polarized epithelium-sperm co-culture system reveals stimulatory factors for the secretion of mouse epididymal quiescin sulfhydryl oxidase 1 极化上皮-精子共培养系统揭示小鼠附睾奎叶素巯基氧化酶1分泌的刺激因素

The Journal of Reproduction and Development Pub Date : 2022-02-26 DOI: 10.1262/jrd.2021-128

Yu-Syuan Wei, W. Lin, Tse-En Wang, Wei-Yun Lee, Sheng-Hsiang Li, F. Lin, B. Nixon, P. Sipilä, P. Tsai

{"title":"Polarized epithelium-sperm co-culture system reveals stimulatory factors for the secretion of mouse epididymal quiescin sulfhydryl oxidase 1","authors":"Yu-Syuan Wei, W. Lin, Tse-En Wang, Wei-Yun Lee, Sheng-Hsiang Li, F. Lin, B. Nixon, P. Sipilä, P. Tsai","doi":"10.1262/jrd.2021-128","DOIUrl":"https://doi.org/10.1262/jrd.2021-128","url":null,"abstract":"Spermatozoa acquire fertilization ability through post-translational modifications. These membrane surface alterations occur in various segments of the epididymis. Quiescin sulfhydryl oxidases, which catalyze thiol-oxidation reactions, are involved in disulfide bond formation, which is essential for sperm maturation, upon transition and migration in the epididymis. Using castration and azoospermia transgenic mouse models, in the present study, we showed that quiescin sulfhydryl oxidase 1 (QSOX1) protein expression and secretion are positively correlated with the presence of testosterone and sperm cells. A two-dimensional in vitro epithelium-sperm co-culture system provided further evidence in support of the notion that both testosterone and its dominant metabolite, 5α-dihydrotestosterone, promote epididymal QSOX1 secretion. We also demonstrated that immature caput spermatozoa, but not mature cauda sperm cells, exhibited great potential to stimulate QSOX1 secretion in vitro, suggesting that sperm maturation is a key regulatory factor for mouse epididymal QSOX1 secretion. Proteomic analysis identified 582 secretory proteins from the co-culture supernatant, of which 258 were sperm-specific and 154 were of epididymal epithelium-origin. Gene Ontology analysis indicated that these secreted proteins exhibit functions known to facilitate sperm membrane organization, cellular activity, and sperm-egg recognition. Taken together, our data demonstrated that testosterone and sperm maturation status are key regulators of mouse epididymal QSOX1 protein expression and secretion.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"135 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122707865","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Effect of autophagy induction and cathepsin B inhibition on developmental competence of poor quality bovine oocytes. 诱导自噬和组织蛋白酶B抑制对劣质牛卵母细胞发育能力的影响。

IF 1.8

The Journal of Reproduction and Development Pub Date : 2020-02-14 Epub Date: 2019-12-25 DOI: 10.1262/jrd.2019-123

Jianye Li, Ahmed Zaky Balboula, Mansour Aboelenain, Takashi Fujii, Satoru Moriyasu, Hanako Bai, Manabu Kawahara, Masashi Takahashi

{"title":"Effect of autophagy induction and cathepsin B inhibition on developmental competence of poor quality bovine oocytes.","authors":"Jianye Li, Ahmed Zaky Balboula, Mansour Aboelenain, Takashi Fujii, Satoru Moriyasu, Hanako Bai, Manabu Kawahara, Masashi Takahashi","doi":"10.1262/jrd.2019-123","DOIUrl":"https://doi.org/10.1262/jrd.2019-123","url":null,"abstract":"The present study investigated the effect of autophagy induction and cathepsin B (CTSB) inhibition on developmental competence of poor quality oocytes. Bovine cumulus oocyte complexes (COCs) were classified as good or poor according to their morphology. Autophagy activity was detected in good and poor germinal vesicle (GV) oocytes. Then E-64, a CTSB inhibitor, rapamycin (Rapa), an autophagy inducer, and combined administration was achieved during invitro maturation (IVM) of poor quality COCs followed by detection of autophagy activity. In the next experiment, E-64, Rapa, and E64 + Rapa, were added during IVM to good and poor quality COCs followed by invitro fertilization and culture for 8 days to investigate whether inhibition of CTSB and/or induction of autophagy improve embryonic development and quality. Autophagy activity was significantly lower in poor quality GV oocytes than in good quality ones. E-64, Rapa and E-64 + Rapa treatment during IVM significantly increased autophagy activity in poor quality oocytes. Addition of Rapa in good quality COCs did not increase the blastocyst rate, whereas E-64 increased the blastocyst rate and total cell number (TCN) with decreasing TUNEL-positive cells. In contrast, Rapa treatment in poor quality COCs significantly increased the blastocyst rate and TCN with decreasing TUNEL-positive cells. These results indicate oocyte quality has different responses to intracellular autophagy induction and CTSB activity control by potential autophagy and catabolic status, however, synergetic effect of autophagy induction and CTSB inhibition can increase developmental competence of both good and poor quality COCs, especially rescue effect in poor quality COCs.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":" ","pages":"83-91"},"PeriodicalIF":1.8,"publicationDate":"2020-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1262/jrd.2019-123","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37489901","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 12

Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA. 利用马PLCζ mRNA从失活精子中产生小鼠后代。

IF 1.8

The Journal of Reproduction and Development Pub Date : 2020-02-14 Epub Date: 2019-12-18 DOI: 10.1262/jrd.2019-043

Yunosuke Yamamoto, Naoki Hirose, Satoshi Kamimura, Sayaka Wakayama, Junya Ito, Masatoshi Ooga, Teruhiko Wakayama

{"title":"Production of mouse offspring from inactivated spermatozoa using horse PLCζ mRNA.","authors":"Yunosuke Yamamoto, Naoki Hirose, Satoshi Kamimura, Sayaka Wakayama, Junya Ito, Masatoshi Ooga, Teruhiko Wakayama","doi":"10.1262/jrd.2019-043","DOIUrl":"https://doi.org/10.1262/jrd.2019-043","url":null,"abstract":"Improving artificial oocyte activation is essential for assisted reproduction or animal biotechnology that can obtain healthy offspring with a high success rate. Here, we examined whether intracytoplasmic injection of equine sperm-specific phospholipase C zeta (ePLCζ) mRNA, the PLCζ with the strongest oocyte activation potential in mammals, could improve the mouse oocyte activation rate and subsequent embryonic development using inactivated spermatozoa. mRNA of mouse PLCζ (mPLCζ) or ePLCζ were injected into mouse oocytes to determine the optimal mRNA concentration to maximize the oocyte activation rate and developmental rate of parthenogenetic embryos in vitro. Full-term development was examined using NaOH-treated inactive spermatozoa using the optimal activation method. We found that the most optimal ePLCζ mRNA concentration was 0.1 ng/µl for mouse oocyte activation, which was ten times stronger than mPLCζ mRNA. The concentration did not affect parthenogenetic embryo development in vitro. Relatively normal blastocysts were obtained with the same developmental rate (52-53% or 48-51%, respectively) when inactive spermatozoa were injected into activated oocytes using ePLCζ or mPLCζ mRNA injection. However, the birth rate after embryo transfer was slightly but significantly decreased in oocytes activated by ePLCζ mRNA (24%) compared to mPLCζ mRNA (37%) or strontium treatment (40%) activation. These results suggest that the higher activation rate does not always correlate the higher birth rate, and some mechanisms might exist in the oocyte activation process that could affect the later developmental stages like full-term development.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":" ","pages":"67-73"},"PeriodicalIF":1.8,"publicationDate":"2020-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1262/jrd.2019-043","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37470932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

Expression of genes encoding mineralocorticoid biosynthetic enzymes and the mineralocorticoid receptor, and levels of mineralocorticoids in the bovine follicle and corpus luteum. 矿化皮质激素生物合成酶和矿化皮质激素受体编码基因的表达及矿化皮质激素在牛卵泡和黄体中的水平。

IF 1.8

The Journal of Reproduction and Development Pub Date : 2020-02-14 Epub Date: 2019-12-15 DOI: 10.1262/jrd.2019-127

Memory Mukangwa, Koki Takizawa, You Aoki, Seizo Hamano, Masafumi Tetsuka

{"title":"Expression of genes encoding mineralocorticoid biosynthetic enzymes and the mineralocorticoid receptor, and levels of mineralocorticoids in the bovine follicle and corpus luteum.","authors":"Memory Mukangwa, Koki Takizawa, You Aoki, Seizo Hamano, Masafumi Tetsuka","doi":"10.1262/jrd.2019-127","DOIUrl":"https://doi.org/10.1262/jrd.2019-127","url":null,"abstract":"Unlike sex steroids, mineralocorticoids have attracted limited attention in ovarian physiology. Recent studies on primates have indicated possible local synthesis and action of mineralocorticoids in the ovary. Here, we examined developmental changes in the levels of mineralocorticoids and expression of genes encoding their biosynthetic enzymes and receptor in the bovine ovary. The follicles and corpora lutea (CL) were collected from F1 heifers. Expression levels of 21α-hydroxylase (CYP21A2), 11β-hydroxylase-1 (CYP11B1), and the mineralocorticoid receptor (NR3C2) in granulosa cells (GC), thecal layers (TL), and CL tissues were quantified by real-time PCR, whereas mineralocorticoids in the follicular fluid were measured by enzyme immunoassay (EIA). TL and GC expressed CYP21A2 and NR3C2, whereas CYP11B1 was expressed at very low or undetectable levels. The expression levels of these genes were not significantly different among small/large and healthy/atretic follicles but were higher in TL than in GC. CYP21A2 and NR3C2 were expressed in all CL stages with higher expression observed in the mid-stage. CYP11B1 expression was only apparent in the mid-stage CL. Aldosterone was detected in all follicles, and its concentration was not significantly different among the follicular groups. In paired large-healthy/atretic follicles, the concentration of deoxycorticosterone, a precursor of aldosterone, was approximately ten-fold higher than that of aldosterone and not significantly different between healthy and atretic follicles. In conclusion, the presence of mineralocorticoids and expression of NR3C2 in the bovine follicle together with the developmental change in the expression of CYP21A2, CYP11B1, and NR3C2 in the CL suggest possible endocrine/paracrine/autocrine roles of mineralocorticoids in the bovine ovary.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":" ","pages":"75-81"},"PeriodicalIF":1.8,"publicationDate":"2020-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1262/jrd.2019-127","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37459225","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10