The Journal of Reproduction and Development最新文献_第7页

γ-Aminobutyric acid suppresses enhancement of hamster sperm hyperactivation by 5-hydroxytryptamine γ-氨基丁酸抑制5-羟色胺增强仓鼠精子过度活化

The Journal of Reproduction and Development Pub Date : 2016-10-24 DOI: 10.1262/jrd.2016-091

M. Fujinoki, Gen L. Takei

引用次数: 11

Involvement of DNA methylation in regulating rat Prop1 gene expression during pituitary organogenesis DNA甲基化参与大鼠垂体器官发生过程中Prop1基因表达的调控

The Journal of Reproduction and Development Pub Date : 2016-10-21 DOI: 10.1262/jrd.2016-102

H. Nishihara, S. Yoshida, Naoko Kanno, Naoto Nishimura, Hiroki Ueharu, J. Ohgane, T. Kato, Y. Kato

{"title":"Involvement of DNA methylation in regulating rat Prop1 gene expression during pituitary organogenesis","authors":"H. Nishihara, S. Yoshida, Naoko Kanno, Naoto Nishimura, Hiroki Ueharu, J. Ohgane, T. Kato, Y. Kato","doi":"10.1262/jrd.2016-102","DOIUrl":"https://doi.org/10.1262/jrd.2016-102","url":null,"abstract":"PROP1 is a pituitary specific transcription factor that plays a crucial role in pituitary organogenesis. The Prop1 shows varied expression patterns that promptly emerge and then fade during the early embryonic period. However, the regulatory mechanisms governing Prop1 expression remain unclear. Here, we investigated whether Prop1 was under epigenetic regulation by DNA methylation. Bisulfite sequencing was performed on DNA obtained from the pituitary glands and livers of rats on embryonic days (E) 13.5 and E14.5, and postnatal days (P) 4 and P30. The methylation of CpG sites in seven regions from 3-kb upstream of the Prop1 transcription start site through to its second intron were examined. Certain differences in CpG-methylation levels were observed in Region-1 (–2772 b to –2355 b), Region-4 (–198 b to +286 b), Region-5 (+671 b to +990 b), and Region-6 (+1113 b to +1273 b) based on comparisons between pituitary and liver DNA on E13.5. DNA methylation in pituitary glands on E14.5, P4, and P30 was generally similar to that observed in in the pituitary gland on E13.5, whereas the anterior and intermediate lobes of the pituitary gland on P4 and P30 showed only small differences. These results indicate that Prop1 is under regulation by CpG methylation during the early period of pituitary primordium development around E13.5.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130052473","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: 1

Efficient mutagenesis by CRISPR/Cas system during meiotic maturation of porcine oocytes 猪卵母细胞减数分裂成熟过程中CRISPR/Cas系统的高效诱变

The Journal of Reproduction and Development Pub Date : 2016-10-21 DOI: 10.1262/jrd.2016-094

Asuka Onuma, W. Fujii, K. Sugiura, K. Naito

{"title":"Efficient mutagenesis by CRISPR/Cas system during meiotic maturation of porcine oocytes","authors":"Asuka Onuma, W. Fujii, K. Sugiura, K. Naito","doi":"10.1262/jrd.2016-094","DOIUrl":"https://doi.org/10.1262/jrd.2016-094","url":null,"abstract":"Genome editing using the CRISPR/Cas system can induce mutations with high efficiency, and allows easier production of genome-modified animals than that offered by the conventional method where embryonic stem cells are used. However, studies using CRISPR/Cas systems have been mostly limited to proliferating somatic cells and pronuclear-stage fertilized eggs. In contrast, the efficiency of a CRISPR/Cas system in immature and maturing oocytes progressing through meiosis has not yet been assessed. In the present study, we evaluated the genome-modification efficiency of the CRISPR/Cas system during meiotic maturation of porcine oocytes. Additionally, the localization of the Cas9 protein in immature oocytes was analyzed in relation to nuclear transport and mutation induction. The results showed that CRISPR/Cas induced mutation with high efficiency even in maturing oocytes with condensed chromosomes, whereas mutations were not induced in GV-stage oocytes. The localization analysis of enhanced green fluorescent protein (EGFP)-tagged Cas9 (Cas9-EGFP) revealed that the nuclei contained lesser Cas9 than the cytoplasm in immature oocytes. Treatment with leptomycin B, a nuclear export inhibitor, increased the amount of nuclear Cas9 and enabled mutation induction in GV oocytes. Our results suggest that CRISPR/Cas systems can be applied to oocytes during meiotic maturation and be implemented in novel applications targeting female genomes.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"120 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116617996","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: 9

An immortalized steroidogenic goat granulosa cell line as a model system to study the effect of the endoplasmic reticulum (ER)-stress response on steroidogenesis 以永生化山羊颗粒细胞系为模型系统，研究内质网应激反应对类固醇生成的影响

The Journal of Reproduction and Development Pub Date : 2016-10-15 DOI: 10.1262/jrd.2016-111

Diqi Yang, Lei Wang, P. Lin, Tingting Jiang, Nan Wang, Fan Zhao, Huatao Chen, Keqiong Tang, Dong Zhou, Aihua Wang, Yaping Jin

{"title":"An immortalized steroidogenic goat granulosa cell line as a model system to study the effect of the endoplasmic reticulum (ER)-stress response on steroidogenesis","authors":"Diqi Yang, Lei Wang, P. Lin, Tingting Jiang, Nan Wang, Fan Zhao, Huatao Chen, Keqiong Tang, Dong Zhou, Aihua Wang, Yaping Jin","doi":"10.1262/jrd.2016-111","DOIUrl":"https://doi.org/10.1262/jrd.2016-111","url":null,"abstract":"With granulosa and theca cells, the ovaries are responsible for producing oocytes and secreting sex steroids such as estrogen and progesterone. Endoplasmic reticulum stress (ERS) plays an important role in follicle atresia and embryo implantation. In this study, goat granulosa cells were isolated from medium-sized (4–6 mm) healthy follicles. Primary granulosa cells were immortalized by transfection with human telomerase reverse transcriptase (hTERT) to establish a goat granulosa cell line (hTERT-GGCs). These hTERT-GGCs expressed hTERT and had relatively long telomeres at passage 50. Furthermore, hTERT-GGCs expressed the gonadotropin receptor genes CYP11A1, StAR, and CYP19A1, which are involved in steroidogenesis. Additionally, progesterone was detectable in hTERT-GGCs. Although the proliferation potential of hTERT-GGCs significantly improved, there was no evidence to suggest that the hTERT-GGCs are tumorigenic. In addition, thapsigargin (Tg) treatment led to a significant dose-dependent decrease in progesterone concentration and steroidogenic enzyme expression. In summary, we successfully generated a stable goat granulosa cell line. We found that Tg induced ERS in hTERT-GGCs, which reduced progesterone production and steroidogenic enzyme expression. Future studies may benefit from using this cell line as a model to explore the molecular mechanisms regulating steroidogenesis and apoptosis in goat granulosa cells.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"661 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134167793","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: 19

Production of α1,3-galactosyltransferase and cytidine monophosphate-N-acetylneuraminic acid hydroxylase gene double-deficient pigs by CRISPR/Cas9 and handmade cloning 利用CRISPR/Cas9和手工克隆制备α1,3-半乳糖转移酶和胞苷单磷酸- n -乙酰神经氨酸羟化酶基因双缺陷猪

The Journal of Reproduction and Development Pub Date : 2016-10-08 DOI: 10.1262/jrd.2016-079

Hanchao Gao, Chengjiang Zhao, Xi Xiang, Yong Li, Yanli Zhao, Zesong Li, D. Pan, Yifan Dai, H. Hara, D. Cooper, Z. Cai, Lisha Mou

{"title":"Production of α1,3-galactosyltransferase and cytidine monophosphate-N-acetylneuraminic acid hydroxylase gene double-deficient pigs by CRISPR/Cas9 and handmade cloning","authors":"Hanchao Gao, Chengjiang Zhao, Xi Xiang, Yong Li, Yanli Zhao, Zesong Li, D. Pan, Yifan Dai, H. Hara, D. Cooper, Z. Cai, Lisha Mou","doi":"10.1262/jrd.2016-079","DOIUrl":"https://doi.org/10.1262/jrd.2016-079","url":null,"abstract":"Gene-knockout pigs hold great promise as a solution to the shortage of organs from donor animals for xenotransplantation. Several groups have generated gene-knockout pigs via clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) and somatic cell nuclear transfer (SCNT). Herein, we adopted a simple and micromanipulator-free method, handmade cloning (HMC) instead of SCNT, to generate double gene-knockout pigs. First, we applied the CRISPR/Cas9 system to target α1,3-galactosyltransferase (GGTA1) and cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) genes simultaneously in porcine fetal fibroblast cells (PFFs), which were derived from wild-type Chinese domestic miniature Wuzhishan pigs. Cell colonies were obtained by screening and were identified by Surveyor assay and sequencing. Next, we chose the GGTA1/CMAH double-knockout (DKO) cells for HMC to produce piglets. As a result, we obtained 11 live bi-allelic GGTA1/CMAH DKO piglets with the identical phenotype. Compared to cells from GGTA1-knockout pigs, human antibody binding and antibody-mediated complement-dependent cytotoxicity were significantly reduced in cells from GGTA1/CMAH DKO pigs, which demonstrated that our pigs would exhibit reduced humoral rejection in xenotransplantation. These data suggested that the combination of CRISPR/Cas9 and HMC technology provided an efficient and new strategy for producing pigs with multiple genetic modifications.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"81 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134343289","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: 41

Efficient pig ICSI using Percoll-selected spermatozoa; evidence for the essential role of phospholipase C-ζ in ICSI success 利用percoll选择精子的高效猪ICSI磷脂酶C-ζ在ICSI成功中的重要作用的证据

The Journal of Reproduction and Development Pub Date : 2016-09-30 DOI: 10.1262/jrd.2016-103

M. Nakai, Shun'ichi Suzuki, J. Ito, D. Fuchimoto, S. Sembon, J. Noguchi, A. Onishi, N. Kashiwazaki, K. Kikuchi

{"title":"Efficient pig ICSI using Percoll-selected spermatozoa; evidence for the essential role of phospholipase C-ζ in ICSI success","authors":"M. Nakai, Shun'ichi Suzuki, J. Ito, D. Fuchimoto, S. Sembon, J. Noguchi, A. Onishi, N. Kashiwazaki, K. Kikuchi","doi":"10.1262/jrd.2016-103","DOIUrl":"https://doi.org/10.1262/jrd.2016-103","url":null,"abstract":"In pigs, the damaged sperm membrane leads to leakage of phospholipase C-ζ (PLCζ), which has been identified as a sperm factor, and a reduction of oocyte-activating ability. In this study, we investigated whether sperm selected by Percoll gradient centrifugation (Percoll) have sufficient PLCζ, and whether the efficiency of fertilization and blastocyst formation after intracytoplasmic sperm injection (ICSI) using Percoll-selected sperm can be improved. Percoll-selected sperm (Percoll group) or sperm without Percoll selection (Control group) were used. A proportion of the oocytes injected with control sperm were subjected to electrical stimulation at 1 h after ICSI (Cont + ES group). It was found that the Percoll group showed a large amount of PLCζ in comparison with the Control group. Furthermore, application of Percoll-selected sperm for ICSI increased the efficiency of fertilization and embryo development. Thus, these results indicate the Percoll-selected sperm have sufficient PLCζ and high oocyte-activating ability after ICSI in pigs.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"11 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132088910","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: 7

Lack of calcium oscillation causes failure of oocyte activation after intracytoplasmic sperm injection in pigs 在猪卵浆内单精子注射后，缺乏钙振荡导致卵母细胞活化失败

The Journal of Reproduction and Development Pub Date : 2016-09-30 DOI: 10.1262/jrd.2016-113

M. Nakai, J. Ito, Shun'ichi Suzuki, D. Fuchimoto, S. Sembon, Misae Suzuki, J. Noguchi, H. Kaneko, A. Onishi, N. Kashiwazaki, K. Kikuchi

{"title":"Lack of calcium oscillation causes failure of oocyte activation after intracytoplasmic sperm injection in pigs","authors":"M. Nakai, J. Ito, Shun'ichi Suzuki, D. Fuchimoto, S. Sembon, Misae Suzuki, J. Noguchi, H. Kaneko, A. Onishi, N. Kashiwazaki, K. Kikuchi","doi":"10.1262/jrd.2016-113","DOIUrl":"https://doi.org/10.1262/jrd.2016-113","url":null,"abstract":"In pigs, the efficiency of embryo production after intracytoplasmic sperm injection (ICSI) is still low because of frequent failure of normal fertilization, which involves formation of two polar bodies and two pronuclei. To clarify the reasons for this, we hypothesized that ICSI does not properly trigger sperm-induced fertilization events, especially intracellular Ca2+ signaling, also known as Ca2+ oscillation. We also suspected that the use of in vitro-matured oocytes might negatively affect fertilization events and embryonic development of sperm-injected oocytes. Therefore, we compared the patterns of Ca2+ oscillation, the efficiency of oocyte activation and normal fertilization, and embryo development to the blastocyst stage among in vivo- or in vitro-matured oocytes after ICSI or in vitro fertilization (IVF). Unexpectedly, we found that the pattern of Ca2+ oscillation, such as the frequency and amplitude of Ca2+ rises, in oocytes after ICSI was similar to that in oocytes after IVF, irrespective of the oocyte source. However, half of the oocytes failed to become activated after ICSI and showed no Ca2+ oscillation. Moreover, the embryonic development of normal fertilized oocytes was reduced when in vitro-matured oocytes were used, irrespective of the fertilization method employed. These findings suggest that low embryo production efficiency after ICSI is attributable mainly to poor developmental ability of in vitro-matured oocytes and a lack of Ca2+ oscillation, rather than the pattern of oscillation.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"30 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124895946","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: 8

Meiotic cohesin subunits RAD21L and REC8 are positioned at distinct regions between lateral elements and transverse filaments in the synaptonemal complex of mouse spermatocytes 小鼠精母细胞突触复合体中减数分裂内聚蛋白亚基RAD21L和REC8分别位于外侧元件和横向细丝之间的不同区域

The Journal of Reproduction and Development Pub Date : 2016-09-26 DOI: 10.1262/jrd.2016-127

M. Rong, A. Matsuda, Y. Hiraoka, Jibak Lee

{"title":"Meiotic cohesin subunits RAD21L and REC8 are positioned at distinct regions between lateral elements and transverse filaments in the synaptonemal complex of mouse spermatocytes","authors":"M. Rong, A. Matsuda, Y. Hiraoka, Jibak Lee","doi":"10.1262/jrd.2016-127","DOIUrl":"https://doi.org/10.1262/jrd.2016-127","url":null,"abstract":"Cohesins containing a meiosis-specific α-kleisin subunit, RAD21L or REC8, play roles in diverse aspects of meiotic chromosome dynamics including formation of axial elements (AEs), assembly of the synaptonemal complex (SC), recombination of homologous chromosomes (homologs), and cohesion of sister chromatids. However, the exact functions of individual α-kleisins remain to be elucidated. Here, we examined the localization of RAD21L and REC8 within the SC by super-resolution microscopy, 3D-SIM. We found that both RAD21L and REC8 were localized at the connection sites between lateral elements (LEs) and transverse filaments (TFs) of pachynema with RAD21L locating interior to REC8 sites. RAD21L and REC8 were not symmetrical in terms of synaptic homologs, suggesting that the arrangement of different cohesins is not strictly fixed along all chromosome axes. Intriguingly, some RAD21L signals, but not REC8 signals, were observed between unsynapsed regions of AEs of zygonema as if they formed a bridge between homologs. Furthermore, the signals of recombination intermediates overlapped with those of RAD21L to a greater degree than with those of REC8. These results highlight the different properties of two meiotic α-kleisins, and strongly support the previous proposition that RAD21L is an atypical cohesin that establishes the association between homologs rather than sister chromatids.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"26 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125186461","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: 23

Adenylation by testis-specific cytoplasmic poly(A) polymerase, PAPOLB/TPAP, is essential for spermatogenesis 睾丸特异性细胞质聚(A)聚合酶(PAPOLB/TPAP)的腺苷化对精子发生至关重要

The Journal of Reproduction and Development Pub Date : 2016-09-18 DOI: 10.1262/jrd.2016-116

S. Kashiwabara, Satsuki Tsuruta, Keitaro Okada, Yutaro Yamaoka, T. Baba

引用次数: 11

Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms 密度梯度离心与游动法相结合，可有效减少形态异常精子

The Journal of Reproduction and Development Pub Date : 2016-09-11 DOI: 10.1262/jrd.2016-112

M. Yamanaka, K. Tomita, S. Hashimoto, Hiroshi Matsumoto, M. Satoh, H. Kato, Y. Hosoi, M. Inoue, Y. Nakaoka, Y. Morimoto

{"title":"Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms","authors":"M. Yamanaka, K. Tomita, S. Hashimoto, Hiroshi Matsumoto, M. Satoh, H. Kato, Y. Hosoi, M. Inoue, Y. Nakaoka, Y. Morimoto","doi":"10.1262/jrd.2016-112","DOIUrl":"https://doi.org/10.1262/jrd.2016-112","url":null,"abstract":"Density gradient centrifugation (DGC) and swim-up techniques have been reported for semen preparation in assisted reproductive techniques in humans. We investigated whether semen preparation using a combination of DGC and swim-up techniques could effectively decrease morphologically abnormal human sperms at the ultrastructural level. Semen samples were obtained from 16 infertile males and fractionated by swim-up following DGC. Ultrastructural abnormalities of sperms obtained from original semen, lower layer of swim-up following DGC, and upper layer of swim-up following DGC were analyzed by transmission electron microscopy. The correlation among ultrastructural head abnormality in sperms from the upper layer of swim-up, fertilization in in vitro fertilization, and pregnancy after embryo transfer was also investigated. Furthermore, sperms with DNA fragmentation in the samples processed via a combination of DGC and swim-up was assessed in a sperm chromatin structure assay. Ultrastructural abnormalities in sperm heads and tails in the upper layer after swim-up following DGC was the lowest among the three groups. Sperms with nuclear vacuoles were the most difficult to eliminate using a combination of DGC and swim-up in all types of head abnormalities. A negative correlation was confirmed between the fertilization rates of intracytoplasmic sperm injection and head abnormality of sperms obtained from the upper layer of the swim-up following DGC. Sperms with DNA fragmentation were effectively decreased using the combination of two techniques. In conclusion, the combination of DGC and swim-up effectively decreased the number of sperms with ultrastructural abnormalities both in the head and in the tail. However, sperms with ultrastructural abnormalities that cannot be completely decreased using a combination of DGC and swim-up may impair fertilization in some cases of intracytoplasmic sperm injection.","PeriodicalId":416064,"journal":{"name":"The Journal of Reproduction and Development","volume":"4 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132264415","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: 29