Molecular Reproduction and Development最新文献

{"title":"Effects of BAMBI on luteinized follicular granulosa cell proliferation and steroid hormone production in sheep","authors":"Yaqi Zhang,&nbsp;Zeyuan Guo,&nbsp;Zhangsheng Du,&nbsp;Zhichao Yao,&nbsp;Tong Guo,&nbsp;Yin Cheng,&nbsp;Kai Wang,&nbsp;Xiaoyan Ma,&nbsp;Chunlu Chen,&nbsp;Ermias Kebreab,&nbsp;Dong Wang,&nbsp;Lihua Lyu","doi":"10.1002/mrd.23674","DOIUrl":"10.1002/mrd.23674","url":null,"abstract":"<p>Bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI) regulates mammalian ovarian follicle growth and maturation; however, its effect on luteinized granulosa cells (LGCs) in sheep ovarian follicles remains unknown. Here we explored the regulatory role of LGC functions and steroid hormone synthesis by BAMBI. Multiple sequence alignment revealed that the sheep <i>BAMBI</i> gene sequence was relatively conserved. Sheep LGCs were strongly positive for BAMBI. LGC proliferation increased when <i>BAMBI</i> was silenced and decreased when <i>BAMBI</i> was overexpressed. After <i>BAMBI</i> overexpression, the expression of <i>CASP3</i>, <i>CASP8</i>, <i>CASP9</i>, and <i>BAX</i> significantly increased, whereas that of <i>BCL2</i> and the ratio of <i>BCL2</i>/<i>BAX</i> expression decreased. The opposite was observed after <i>BAMBI</i> silencing. <i>CDKN1A, CCND1</i>, and <i>CCND2</i> were downregulated with <i>BAMBI</i> overexpression and upregulated with <i>BAMBI</i> silencing. Expression of steroid hormone-related genes (<i>CYP11A1</i>, <i>STAR</i>, and <i>3BHSD</i>), except <i>CYP19A1</i>, significantly increased after <i>BAMBI</i> overexpression. Moreover, estrogen and progesterone secretion increased after <i>BAMBI</i> overexpression and decreased after <i>BAMBI</i> interference. The effect of the exogenous addition of bone morphogenetic protein 2 (BMP2) on GCs was similar to that of <i>BAMBI</i> overexpression. In conclusion, BAMBI can regulate the proliferation and steroid hormone synthesis of sheep LGCs, and BMP2 can affect LGCs as an activator of <i>BAMBI</i>. These findings provide a basis for further research on the physiological role of BAMBI.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 3","pages":"153-165"},"PeriodicalIF":2.5,"publicationDate":"2023-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9676323","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CRISPR/Cas9 knockin methodology for the sea urchin embryo","authors":"Nathalie Oulhen,&nbsp;Shumpei Morita,&nbsp;Jacob F. Warner,&nbsp;Gary Wessel","doi":"10.1002/mrd.23672","DOIUrl":"10.1002/mrd.23672","url":null,"abstract":"CRISPR technology has revolutionized the biological research world, making animals heretofore recalcitrant to genetic manipulation, accessible to analysis of specific gene functions. Building upon the demonstration of targeted gene mutations in the sea urchin (CRISPR knockout) (Fleming et al., 2021; Lin et al., 2019; Lin & Su, 2016; Liu et al., 2019; Vyas et al., 2022), investigators may now be able to insert exogenous DNA into specific locations in the genome (CRISPR knockin). Such Cas9‐mediated knockins will reveal sites of gene expression, and function. By judicious selection of exogenously encoded tags, for example, a fluorescent reporter, an investigator may then follow specific gene activities and cell lineages throughout development in live embryos. This tag can also be used for protein pull‐down without requiring an antibody for the targeted protein. Here we describe a procedure for CRISPR‐based knock‐in DNA in the sea urchin Strongylocentrotus purpuratus. Sea urchin larvae produce echinochrome pigments that require several gene functions including the enzyme polyketide synthase 1 (PKS1) (Barsi et al., 2015; Calestani & Wessel, 2018; Calestani et al., 2003; Perillo et al., 2020; Wessel et al., 2020). Sp PKS1 expression is restricted to a small population of ∼50 cells of theVeg2 lineage of the animal (Barsi et al., 2015; Calestani et al., 2003). We realized that using PKS1 to evaluate CRISPR knockin success was highly stringent since the insertion must occur within that small lineage, and be expressed by yet a smaller population of the lineage. Mutations of the gene encoding PKS1 by CRISPR knockout resulted in albino larvae, an easy phenotype to assess with simple brightfield microscopy (Oulhen & Wessel, 2016a). A single gRNA was previously shown to mutate PKS1 by Cas9 activity, nearly 100% of the time in embryos from S. purpuratus and Hemicentrotus pulcherrimus (Liu et al., 2019; Oulhen & Wessel, 2016a; Oulhen et al., 2022). We took advantage of this highly efficient gRNA to test and to optimize Cas9‐ mediated methodology in the sea urchin S. purpuratus. We tested three different donor templates for their efficacy in selectively knocking‐in exogenous DNA encoding a fluorescent protein: plasmid DNA, linear double‐stranded DNA, single stranded DNA. The key for this test is a highly efficient gRNA against the target gene, and a DNA repair template that contains homologous regions to the target sequence for homology directed repair (Figure S1). Investigators have previously injected linear DNA into sea urchin eggs/early embryos, which results in rapid and extensive concatenation (McMahon et al., 1985) that appears to be detrimental to high‐fidelity insertion (data not shown). To counter this concern, we tested circular plasmid‐based strategies. Here, the DNA repair template targeting the cleaved genomic locus was contained within a plasmid and was accessible for insertion before or following CRISPR‐Cas9 cutting of the same flanking sequence i","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 2","pages":"69-72"},"PeriodicalIF":2.5,"publicationDate":"2023-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.23672","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9558324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Table of Contents, Volume 90, Issue 1, January 2023","authors":"","doi":"10.1002/mrd.23580","DOIUrl":"10.1002/mrd.23580","url":null,"abstract":"","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 1","pages":"1"},"PeriodicalIF":2.5,"publicationDate":"2023-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.23580","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42101396","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Front Cover Image, Volume 90, Issue 1, January 2023","authors":"Kohei Umezu,&nbsp;Irina V. Larina","doi":"10.1002/mrd.23673","DOIUrl":"10.1002/mrd.23673","url":null,"abstract":"<p>Cover Caption: The cover image is based on the Review Article <i>Optical coherence tomography for dynamic investigation of mammalian reproductive processes</i> by Kohei Umezu et al., https://doi.org/10.1002/mrd.23665.\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure></p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 1","pages":"i"},"PeriodicalIF":2.5,"publicationDate":"2023-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.23673","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47826569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Iberiotoxin and clofilium regulate hyperactivation, acrosome reaction, and ion homeostasis synergistically during human sperm capacitation","authors":"Yayan Wang,&nbsp;Tian Gao,&nbsp;Lijun Shan,&nbsp;Kun Li,&nbsp;Fei Liang,&nbsp;Jianmin Yu,&nbsp;Ya Ni,&nbsp;Peibei Sun","doi":"10.1002/mrd.23671","DOIUrl":"10.1002/mrd.23671","url":null,"abstract":"<p>Potassium channels play essential roles in the regulation of male fertility. However, potassium channels mediating K⁺ currents in human sperm (I_KSper) remain controversial. Besides SLO3, the SLO1 potassium channel is a potential candidate for human sperm KSper. This study intends to elucidate the function of SLO1 potassium channel during human sperm capacitation. Human sperm were treated with iberiotoxin (IbTX, a SLO1 specific inhibitor) and clofilium (SLO3 inhibitor) separately or simultaneously during in vitro capacitation. A computer-assisted sperm analyzer was used to assess sperm motility. The sperm acrosome reaction (AR) was analyzed using fluorescein isothiocyanate-conjugated <i>Pisum sativum</i> agglutinin staining. Sperm protein tyrosine phosphorylation was studied using western blotting. Intracellular Ca²⁺, K⁺, Cl⁻, and pH were analyzed using ion fluorescence probes. Independent inhibition with IbTX or clofilium decreased the sperm hyperactivation, AR, and protein tyrosine phosphorylation, and was accompanied by an increase in [K⁺]_i, [Cl⁻]_i, and pH_i, but a decrease in [Ca²⁺]_i. Simultaneously inhibition with IbTX and clofilium lower sperm hyperactivation and AR more than independent inhibition. The increase in [K⁺]_i, [Cl⁻]_i, and pH_i, and the decrease in [Ca²⁺]_i were more pronounced. This study suggested that the SLO1 potassium channel may have synergic roles with SLO3 during human sperm capacitation.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 3","pages":"129-140"},"PeriodicalIF":2.5,"publicationDate":"2023-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9313704","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Scrambled eggs—Negative impacts of heat stress and chemical exposures on ovarian function in swine","authors":"M. Estefanía González-Alvarez,&nbsp;Crystal M. Roach,&nbsp;Aileen F. Keating","doi":"10.1002/mrd.23669","DOIUrl":"10.1002/mrd.23669","url":null,"abstract":"<div>\u0000 \u0000 <p>Exposure to environmental toxicants and hyperthermia can hamper reproduction in female mammals including swine. Phenotypic manifestations include poor quality oocytes, endocrine disruption, infertility, lengthened time to conceive, pregnancy loss, and embryonic defects. The ovary has the capacity for toxicant biotransformation, regulated in part by the phosphatidylinositol-3 kinase signaling pathway. The impacts of exposure to mycotoxins and pesticides on swine reproduction and the potential for an emerging chemical class of concern, the per- and polyfluoroalkylated substances, to hamper porcine reproduction are reviewed. The negative impairments of heat stress (HS) on swine reproductive outcomes are also described and the cumulative effect of environmental exposures, such as HS, when present in conjunction with a toxicant is considered.</p></div>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 7","pages":"503-516"},"PeriodicalIF":2.5,"publicationDate":"2023-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.23669","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10231814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular vesicles: Focus on peri-implantation period of pregnancy in pigs","authors":"Maria M. Guzewska,&nbsp;Joanna Szuszkiewicz,&nbsp;Monika M. Kaczmarek","doi":"10.1002/mrd.23664","DOIUrl":"10.1002/mrd.23664","url":null,"abstract":"<p>The establishment of cell-to-cell communication between the endometrium and the developing embryo is the most important step in successful mammalian pregnancy. Close interaction between the uterine luminal epithelium and trophoblast cells requires triggering timely molecular dialog for successful maternal recognition of pregnancy, embryo implantation, and placenta development. Quite recently, extracellular vesicles (EVs) carrying unique molecular cargo emerged as evolutionarily conserved mediators of cell-to-cell communication during early pregnancy. To date, the presence of EVs at the embryo–maternal interface has been demonstrated in numerous mammals, including domestic livestock, such as pigs. However, few studies have focused on revealing the mechanism of EV-mediated crosstalk between developing early embryos and receptive endometrium. Over the past years, it has appeared that understanding the role of EVs in mammalian reproduction can substantially improve our understanding of the biological challenges of successful reproductive performance. This review describes current knowledge of EVs, specifically in relation to the peri-implantation period in pigs, characterized by common features of embryo implantation and high embryonic mortality in mammals.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 7","pages":"634-645"},"PeriodicalIF":2.5,"publicationDate":"2023-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.23664","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10528676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deterioration of mitochondrial biogenesis and degradation in the endometrium is a cause of subfertility in cows","authors":"Shuichi Matsuyama,&nbsp;Sho Nakamura,&nbsp;Shiori Minabe,&nbsp;Miki Sakatani,&nbsp;Naoki Takenouchi,&nbsp;Takuya Sasaki,&nbsp;Yuki Inoue,&nbsp;Hisataka Iwata,&nbsp;Koji Kimura","doi":"10.1002/mrd.23670","DOIUrl":"10.1002/mrd.23670","url":null,"abstract":"<p>To investigate possible causes of reproductive failure, we conducted global endometrial gene expression analyses in fertile and subfertile cows. Ingenuity pathway analysis showed that RICTOR and SIRT3 are significant upstream regulators for highly expressed genes in fertile cows, and are predicted to be activated upstream regulators of normal mitochondrial respiration. Canonical pathway analysis revealed that these highly expressed genes are involved in the activation of mitochondrial oxidative phosphorylation. Therefore, in subfertile cows, the inactivation of RICTOR and SIRT3 may correlate with decreased capacity of mitochondrial respiration. Furthermore, the expression levels of most mitochondrial DNA genes and nuclear genes encoding mitochondrial proteins were higher in subfertile cows. The mitochondrial DNA copy number was significantly higher in the endometrium of subfertile cows, whereas the ATP content did not differ between fertile and subfertile cows. Quantitative reverse transcription-PCR analysis demonstrated that the expression of <i>PGC1a, TFAM, MFN1, FIS1</i>, and <i>BCL2L13</i> were significantly lower in subfertile cows. In addition, transmission electron microscopy images showed mitochondrial swelling in the endometrial cells of the subfertile cow. These results suggest that poor-quality mitochondria accumulate in the endometrium owing to a reduced capacity for mitochondrial biogenesis, fusion, fission, and degradation in subfertile cows, and may contribute to infertility.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 3","pages":"141-152"},"PeriodicalIF":2.5,"publicationDate":"2023-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9313698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"One Health challenges for pig reproduction","authors":"Olli Peltoniemi,&nbsp;Topi Tanskanen,&nbsp;Maria Kareskoski","doi":"10.1002/mrd.23666","DOIUrl":"10.1002/mrd.23666","url":null,"abstract":"<p>The current state of the world challenges pig reproduction as an important part of One Health, which involves interrelationships between animal, human and environmental health. The One Health concept underlines a comparative aspect in reproductive physiology and disease occurrence, bridging knowledge from one species to another. Seasonal changes in the environment affect pig reproduction and climate change may further strengthen those effects. Endocrine-disrupting chemicals (EDCs), and specifically phthalates and heavy metals, interfere with endocrine function, and thereby sexual behavior, fertilization capacity and steroidogenesis. Reproductive infections and extended semen storage are important indications for antimicrobial use. Innovative solutions are needed to explore alternatives to antimicrobials. Efforts to ensure reproductive efficiency have prolonged farrowing as litter size has doubled over the past three decades, compromising immune transfer and welfare. Physiological, metabolic and programming related events around parturition are key areas for future One Health research in pig reproduction. In conclusion, climate change challenges reproductive management and breeding. More resilient pigs that can tolerate harsh environment but maintain high reproductive performance are needed. EDCs continue to grow as an environmental challenge for reproductive management and alternatives to antibiotics will be required.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 7","pages":"420-435"},"PeriodicalIF":2.5,"publicationDate":"2023-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/mrd.23666","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10174971","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Circular RNA SESN2 aggravates gestational trophoblast cell damage induced by high glucose by binding to IGF2BP2","authors":"Xin Huang,&nbsp;Linlin Guo","doi":"10.1002/mrd.23667","DOIUrl":"10.1002/mrd.23667","url":null,"abstract":"<p>Gestational diabetes mellitus (GDM) is a common disease in pregnant women that threatens maternal and fetal health. Circular RNAs (circRNAs) have been considered potential diagnostic markers for GDM and affect trophoblast cell phenotypes. This study aimed to explore the effect of circSESN2 on high glucose (HG)-treated trophoblast cells. Peripheral blood and placental tissues were taken from patients with GDM, in which circSESN2 and IGF2BP2 levels were detected by quantitative reverse transcription polymerase chain reaction and/or western blot. HTR-8/SVneo cells were treated with 25 mM glucose and transduced with circSESN2 or IGF2BP2 knockdown vectors. HTR-8/SVneo cell viability was evaluated by MTT assay, cell migration by scratch test, and cell invasion by transwell assay, IL-1β, IL-6, TNF-α, malondialdehyde, and superoxide dismutase levels by ELISA or kits, and reactive oxygen species levels by DCFH-DA probes. The binding between circSESN2 and IGF2BP2 was verified by RNA pulldown and RIP assays. CircSESN2 and IGF2BP2 were overexpressed in GDM patients. Suppressing circSESN2 or IGF2BP2 increased HTR-8/SVneo cell invasion and migration, decreased cell apoptosis, and reduced pro-inflammatory cytokine release and oxidative stress injury. CircSESN2 bound IGF2BP2 and IGF2BP2 overexpression accelerated HG-induced HTR-8/SVneo cell damage despite circSESN2 knockdown. Collectively, circSESN2 exacerbated HG-induced trophoblast cell damage by binding IGF2BP2 and upregulating its protein expression.</p>","PeriodicalId":18856,"journal":{"name":"Molecular Reproduction and Development","volume":"90 2","pages":"73-86"},"PeriodicalIF":2.5,"publicationDate":"2023-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9077202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}