Pub Date : 2025-06-01Epub Date: 2025-04-14DOI: 10.1016/j.repbio.2025.101018
Anna Wójtowicz, Agnieszka Sadowska, Katarzyna Piotrowska-Tomala, Anna Szóstek-Mioduchowska
The role of AREG in the development of fibrosis in the progression of endometrosis in mare remains unknown. We aimed to determine the effects of AREG on fibroblast functional characteristics as well as the expression of extracellular matrix (ECM)-associated genes in fibroblast derived from non-fibrotic and fibrotic equine endometria. Our findings suggest that the mechanisms associated with ECM remodeling regulated by AREG in non-fibrotic fibroblasts may be dysregulated in the progression of fibrosis in endometrosis.
{"title":"The effect of amphiregulin on equine endometrial fibroblasts: in vitro responses of fibroblast derived from non-fibrotic and fibrotic endometrium","authors":"Anna Wójtowicz, Agnieszka Sadowska, Katarzyna Piotrowska-Tomala, Anna Szóstek-Mioduchowska","doi":"10.1016/j.repbio.2025.101018","DOIUrl":"10.1016/j.repbio.2025.101018","url":null,"abstract":"<div><div>The role of AREG in the development of fibrosis in the progression of endometrosis in mare remains unknown. We aimed to determine the effects of AREG on fibroblast functional characteristics as well as the expression of extracellular matrix (ECM)-associated genes in fibroblast derived from non-fibrotic and fibrotic equine endometria. Our findings suggest that the mechanisms associated with ECM remodeling regulated by AREG in non-fibrotic fibroblasts may be dysregulated in the progression of fibrosis in endometrosis.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":""},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143825462","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}
Pub Date : 2025-06-01Epub Date: 2025-04-19DOI: 10.1016/j.repbio.2025.101016
Camila Lapuente , Augusto Lantermino , Sol Arioni , Paula G. Blanco , Cristina Gobello
Anti-Müllerian hormone (AMH) is a dimeric glycoprotein that belongs to the transforming growth factor-β (TGF-β) superfamily. This hormone that is produced by gonadal Sertoli cells in males and granulosa cells in females has been extensively studied in humans, rodents, and livestock species. Research on AMH in felids began in 2011 and given the increasing number of studies over recent years, an updated literature review is necessary to clarify and organize future research directions. The objective of this article was to conduct a systematic review of AMH in domestic and wild felids. From a literature search of international publications, 23 were selected for inclusion. AMH determinations were performed using commercial enzyme-linked immunosorbent assays (ELISA) and electrochemiluminescence immunoassays (EQLIA). In female felids, AMH concentrations decrease with age, along with follicular reserve diminution. AMH can also be used to diagnose granulosa cell ovarian tumors and cryptorchidism in females and males, respectively. This hormone serves as a marker for reproductive status and can reflect gonadal function in both genders. Furthermore, AMH may prove to be a valuable predictive tool for reproductive biotechnologies in both domestic and wild felids. Several aspects of this hormone still remain to be elucidated, including its variations throughout the estrous cycle and the effect of photoperiod. Finally, standardization of assays and the establishment of reference ranges for both domestic and wild animals are necessary for widespread clinical application and future research development.
{"title":"Anti-mullerian hormone in felids: A systematic review","authors":"Camila Lapuente , Augusto Lantermino , Sol Arioni , Paula G. Blanco , Cristina Gobello","doi":"10.1016/j.repbio.2025.101016","DOIUrl":"10.1016/j.repbio.2025.101016","url":null,"abstract":"<div><div>Anti-Müllerian hormone (AMH) is a dimeric glycoprotein that belongs to the transforming growth factor-β (TGF-β) superfamily. This hormone that is produced by gonadal Sertoli cells in males and granulosa cells in females has been extensively studied in humans, rodents, and livestock species. Research on AMH in felids began in 2011 and given the increasing number of studies over recent years, an updated literature review is necessary to clarify and organize future research directions. The objective of this article was to conduct a systematic review of AMH in domestic and wild felids. From a literature search of international publications, 23 were selected for inclusion. AMH determinations were performed using commercial enzyme-linked immunosorbent assays (ELISA) and electrochemiluminescence immunoassays (EQLIA). In female felids, AMH concentrations decrease with age, along with follicular reserve diminution. AMH can also be used to diagnose granulosa cell ovarian tumors and cryptorchidism in females and males, respectively. This hormone serves as a marker for reproductive status and can reflect gonadal function in both genders. Furthermore, AMH may prove to be a valuable predictive tool for reproductive biotechnologies in both domestic and wild felids. Several aspects of this hormone still remain to be elucidated, including its variations throughout the estrous cycle and the effect of photoperiod. Finally, standardization of assays and the establishment of reference ranges for both domestic and wild animals are necessary for widespread clinical application and future research development.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101016"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848159","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}
Pub Date : 2025-06-01Epub Date: 2025-03-30DOI: 10.1016/j.repbio.2025.101015
Farha A. Ali Shafi , Ali Thoulfikar A. Imeer , Hassan Ali Abood Nassrullah , Ali Mutashar Naeemah
This study investigates the impact of extracellular glucose concentrations on antioxidant capacity, viability, and microRNA (miR) expression in TM4 Sertoli cell lines. TM4 cells were cultured in high-glucose (115 µm) and low-glucose (<505 µm) conditions to simulate hyperglycemia and glucose starvation, respectively. The study measured total antioxidant capacity (TAC), malondialdehyde (MDA), total oxidant status (TOS), glutathione (GSH), glutathione disulfide (GSSG), NADP/NADPH, glutathione peroxidase (GPX), and glutathione reductase (GR) levels. MiR-17, miR-34, miR-106a, and miR-200a expression levels were assessed. Cell viability and apoptosis were evaluated using MTT assay and acridine-orange staining. Results indicated that high glucose reduced miR-17 expression while low glucose increased it. Both glucose conditions elevated miR-34, miR-106a, and miR-200a expressions. TAC levels decreased, while TOS and MDA levels increased significantly under both conditions. High glucose had no significant effect on GPX and GR levels, whereas low glucose decreased them. Both conditions led to reduced GSH levels, increased GSSG levels, and altered NADP/NADPH ratio. Increased apoptosis and decreased cell viability were observed under both glucose conditions. These findings suggest that extracellular glucose levels significantly dysregulate miRNA expression, antioxidant capacities, and redox buffer systems in TM4 cells. High glucose conditions suppress miR-17 expression, increase miR-34 and miR-106a levels, and induce reductive buffer imbalance. Conversely, low glucose conditions trigger compensatory mechanisms via increased miR-17 expression to enhance antioxidant status while reducing GPX and GR levels. These results provide insights into the molecular responses of Sertoli cells under varying glucose environments, highlighting potential therapeutic pathways for conditions like diabetes and metabolic dysfunctions.
{"title":"The impact of extracellular glucose concentrations on antioxidant capacity, viability, and microRNA expression in TM4 Sertoli cells","authors":"Farha A. Ali Shafi , Ali Thoulfikar A. Imeer , Hassan Ali Abood Nassrullah , Ali Mutashar Naeemah","doi":"10.1016/j.repbio.2025.101015","DOIUrl":"10.1016/j.repbio.2025.101015","url":null,"abstract":"<div><div>This study investigates the impact of extracellular glucose concentrations on antioxidant capacity, viability, and microRNA (miR) expression in TM4 Sertoli cell lines. TM4 cells were cultured in high-glucose (115 µm) and low-glucose (<505 µm) conditions to simulate hyperglycemia and glucose starvation, respectively. The study measured total antioxidant capacity (TAC), malondialdehyde (MDA), total oxidant status (TOS), glutathione (GSH), glutathione disulfide (GSSG), NADP/NADPH, glutathione peroxidase (GPX), and glutathione reductase (GR) levels. MiR-17, miR-34, miR-106a, and miR-200a expression levels were assessed. Cell viability and apoptosis were evaluated using MTT assay and acridine-orange staining. Results indicated that high glucose reduced miR-17 expression while low glucose increased it. Both glucose conditions elevated miR-34, miR-106a, and miR-200a expressions. TAC levels decreased, while TOS and MDA levels increased significantly under both conditions. High glucose had no significant effect on GPX and GR levels, whereas low glucose decreased them. Both conditions led to reduced GSH levels, increased GSSG levels, and altered NADP/NADPH ratio. Increased apoptosis and decreased cell viability were observed under both glucose conditions. These findings suggest that extracellular glucose levels significantly dysregulate miRNA expression, antioxidant capacities, and redox buffer systems in TM4 cells. High glucose conditions suppress miR-17 expression, increase miR-34 and miR-106a levels, and induce reductive buffer imbalance. Conversely, low glucose conditions trigger compensatory mechanisms via increased miR-17 expression to enhance antioxidant status while reducing GPX and GR levels. These results provide insights into the molecular responses of Sertoli cells under varying glucose environments, highlighting potential therapeutic pathways for conditions like diabetes and metabolic dysfunctions.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101015"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734743","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}
Pub Date : 2025-06-01Epub Date: 2025-02-03DOI: 10.1016/j.repbio.2025.100995
Maxim A. Filatov, Leonid A. Ilchuk, Iuliia P. Baikova
Ex utero culture of postimplantation embryos remains one of the unsolved tasks in developmental biology. This technique may be required for infertility treatment, observation of embryo development and assessing the embryotoxicity of certain chemical agents. We describe novel method for E12.5 mouse embryos whole embryo culture system, which maintains embryo viability for 24 h. The culture system is based on the bubbling of pure oxygen through the culture medium. The oxygen was obtained by a chemical method. Each tube containing three embryos held 8 ml of culture medium composed of 6 ml of FBS, 2 ml of DMEM/F12, 80 μl of 40 % glucose and 30 μl of antibiotics (a mixture of penicillin 5000 UI/ml and streptomycin 5000 μg/ml). We observed initiation of auricle formation, as well as progression of eye development. Embryo viability was confirmed by the presence of heartbeat. The ratio of viable embryos after 24 h of culture was 27,78 %. However, many viable embryos exhibited massive hemorrhage attributed to oxygen insufficiency. The described culture system may be useful for the investigation of teratogenic compounds on the development of organs. Nonetheless, it is not suitable for ex utero culture of mouse embryos at more advanced stages due to the fact that embryos at such stages require more oxygen for the development than can be dissolved in the culture medium and consumed by embryos through diffusion. A potential solution to this issue is connecting the embryonic bloodstream to an oxygenator.
移植后胚胎的体外培养是发育生物学中尚未解决的问题之一。这项技术可能需要用于不孕症治疗、胚胎发育观察和评估某些化学制剂的胚胎毒性。我们描述了一种新的E12.5小鼠胚胎全胚培养系统，该系统可维持胚胎活力24 h。培养系统是基于纯氧通过培养基的鼓泡。氧气是用化学方法得到的。每管装有3个胚胎，培养液为8 ml，培养基由6 ml FBS、2 ml DMEM/F12、80 μl 40 %葡萄糖和30 μl抗生素（青霉素5000 UI/ml和链霉素5000 μg/ml混合）组成。我们观察到耳廓形成的开始，以及眼睛发育的进展。胚胎活力通过心跳的存在得到证实。培养24 h后的活胚率为27.78 %。然而，许多存活的胚胎由于缺氧而出现大量出血。所描述的培养体系可能有助于研究致畸化合物对器官发育的影响。然而，它并不适用于更高级阶段的小鼠胚胎的体外培养，因为这些阶段的胚胎需要更多的氧气来发育，而不是溶解在培养基中并通过扩散被胚胎消耗。这个问题的一个潜在解决方案是将胚胎血流与氧合器连接起来。
{"title":"E12.5 whole mouse embryo culture","authors":"Maxim A. Filatov, Leonid A. Ilchuk, Iuliia P. Baikova","doi":"10.1016/j.repbio.2025.100995","DOIUrl":"10.1016/j.repbio.2025.100995","url":null,"abstract":"<div><div><em>Ex utero</em> culture of postimplantation embryos remains one of the unsolved tasks in developmental biology. This technique may be required for infertility treatment, observation of embryo development and assessing the embryotoxicity of certain chemical agents. We describe novel method for E12.5 mouse embryos whole embryo culture system, which maintains embryo viability for 24 h. The culture system is based on the bubbling of pure oxygen through the culture medium. The oxygen was obtained by a chemical method. Each tube containing three embryos held 8 ml of culture medium composed of 6 ml of FBS, 2 ml of DMEM/F12, 80 μl of 40 % glucose and 30 μl of antibiotics (a mixture of penicillin 5000 UI/ml and streptomycin 5000 μg/ml). We observed initiation of auricle formation, as well as progression of eye development. Embryo viability was confirmed by the presence of heartbeat. The ratio of viable embryos after 24 h of culture was 27,78 %. However, many viable embryos exhibited massive hemorrhage attributed to oxygen insufficiency. The described culture system may be useful for the investigation of teratogenic compounds on the development of organs. Nonetheless, it is not suitable for <em>ex utero</em> culture of mouse embryos at more advanced stages due to the fact that embryos at such stages require more oxygen for the development than can be dissolved in the culture medium and consumed by embryos through diffusion. A potential solution to this issue is connecting the embryonic bloodstream to an oxygenator.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 100995"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143168795","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}
The pig industry is interested in increasing the number of female piglets by using sexed semen. Immunological sperm sexing is a promising method. This study investigated and produced a monoclonal antibody (MAbs) targeted to plasma membrane epitopes on porcine Y-chromosome-bearing sperm. Two BALB/c mice were immunized with 92.08 % high-purity porcine Y-sperm, which was separated by a cell sorter flow cytometer. The hybridoma cells were a fusion of myeloma cells (P3X63Ag8.653) and splenocyte cells from immunized mice. Indirect ELISA screening for positive antibodies produced by a single clone well (C2B2) with a high titer specific to porcine Y-sperm. The C2B2 clone was used to produce and purify C2B2-MAbs, yielding 2.78 ± 0.78 µg/mL. The C2B2-MAbs was highly specific to Y-sperm (100.00 %) and had a low cross-reactivity with X-sperm (3.25 %). Therefore, the percentage cross-reactivity of C2B2-MAbs was low for conventional sperm from various livestock, including 0.34 % for Angus, 0.38 % for Holstein-Friesian, 0.20 % for goats, and 0.25 % for buffalo. The bright fluorescence of FITC displayed by the C2B2-MAbs bound to the plasma membrane of porcine Y-sperm provided evidence of affinity between them. However, the C2B2-MAbs bound to an X-sperm lacked fluorescence. C2B2-MAbs showed specificity for the plasma membrane of porcine Y-sperm, which can be used in porcine semen sexing in further studies.
养猪业希望通过使用性别化精液来增加雌性仔猪的数量。免疫学精子性别鉴定是一种很有前景的方法。本研究调查并制备了一种针对猪 Y 染色体精子质膜表位的单克隆抗体（MAbs）。用 92.08% 的高纯度猪 Y-精子对两只 BALB/c 小鼠进行免疫，并用细胞分拣流式细胞仪对其进行分离。杂交瘤细胞是骨髓瘤细胞（P3X63Ag8.653）与免疫小鼠脾细胞的融合。间接 ELISA 筛选单个克隆孔（C2B2）产生的阳性抗体，该抗体对猪 Y-精子具有高滴度特异性。C2B2 克隆用于生产和纯化 C2B2-MAbs，产量为 2.78 ± 0.78 µg/mL。C2B2-MAbs 对 Y-精子的特异性很高（100.00%），与 X-精子的交叉反应性很低（3.25%）。因此，C2B2-MAbs 与各种家畜的常规精子的交叉反应率较低，其中安格斯为 0.34%，荷斯坦-弗里斯兰为 0.38%，山羊为 0.20%，水牛为 0.25%。与猪 Y 型精子质膜结合的 C2B2-MAbs 显示出明亮的 FITC 荧光，这证明了它们之间的亲和性。然而，与 X 精子结合的 C2B2-MAbs 却没有荧光。C2B2-MAbs 对猪 Y 型精子的质膜具有特异性，可用于猪精液性别鉴定的进一步研究。
{"title":"Production of monoclonal antibodies targeting plasma membrane of porcine Y-chromosome-bearing sperm","authors":"Onpreeya Chot , Marninphan Thongkham , Apinya Satsook , Chaiwat Arjin , Supamit Mekchay , Surat Hongsibsong , Korawan Sringarm","doi":"10.1016/j.repbio.2025.101009","DOIUrl":"10.1016/j.repbio.2025.101009","url":null,"abstract":"<div><div>The pig industry is interested in increasing the number of female piglets by using sexed semen. Immunological sperm sexing is a promising method. This study investigated and produced a monoclonal antibody (MAbs) targeted to plasma membrane epitopes on porcine Y-chromosome-bearing sperm. Two BALB/c mice were immunized with 92.08 % high-purity porcine Y-sperm, which was separated by a cell sorter flow cytometer. The hybridoma cells were a fusion of myeloma cells (P3X63Ag8.653) and splenocyte cells from immunized mice. Indirect ELISA screening for positive antibodies produced by a single clone well (C2B2) with a high titer specific to porcine Y-sperm. The C2B2 clone was used to produce and purify C2B2-MAbs, yielding 2.78 ± 0.78 µg/mL. The C2B2-MAbs was highly specific to Y-sperm (100.00 %) and had a low cross-reactivity with X-sperm (3.25 %). Therefore, the percentage cross-reactivity of C2B2-MAbs was low for conventional sperm from various livestock, including 0.34 % for Angus, 0.38 % for Holstein-Friesian, 0.20 % for goats, and 0.25 % for buffalo. The bright fluorescence of FITC displayed by the C2B2-MAbs bound to the plasma membrane of porcine Y-sperm provided evidence of affinity between them. However, the C2B2-MAbs bound to an X-sperm lacked fluorescence. C2B2-MAbs showed specificity for the plasma membrane of porcine Y-sperm, which can be used in porcine semen sexing in further studies.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101009"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143629454","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}
Pub Date : 2025-06-01Epub Date: 2025-03-29DOI: 10.1016/j.repbio.2025.101012
Michela Calanni-Pileri , Joachim M. Weitzel , Andreas Hoeflich , Martina Langhammer , Marten Michaelis
To date, animal models with reproductive phenotypes are knockout or transgenic and typically exhibit reduced fertility or infertility. This limits research to studying single-gene effects or loss of fertility. By contrast, Dummerstorf high-fertility mouse lines 1 and 2 (FL1 and FL2) are two unique outbred selection models that demonstrate exceptional reproductive performance. After approximately 50 years of selection, both lines have doubled the number of ovulated oocytes per cycle and consequently their litter size (>20 vs ∼11) compared to the unselected mice of the same founder population (Dummerstorf FZTDU, ctrl line). FL1 and FL2 exhibit atypical estrous cycle length and altered levels of hormones, such as insulin and leptin, which are associated with GnRH release and/or increased body fat content. Unlike typical cases where these factors impair fertility, they instead contribute to the FLs’ high reproductive capacity: the increased ovulation rate results from an upgrade in the quality of their oocytes, influenced by different ovarian lipid profile. In the present study, we analyzed the expression of IGF-axis marker genes linked to reproductive performance and FL-specific traits in three tissues. We found that lepr, which plays a critical role in implantation, was upregulated in the FL1 uterus (1.5-fold vs. ctrl, p < 0.05). In FL1 follicles, igf1, IGF-biding proteins (IGFBP2, IGFBP4) and hsf1—which is involved in gametogenesis—were significantly upregulated (1–4-fold vs. ctrl, p < 0.05 for igf1, hsf1 and IGFBP4; p < 0.01 for igfbp2). In FL2, uterine size was reduced relatively to the body weight (∼0.2 % FL2 vs. 0.25 % in ctrl and 0.28 % in FL1, p < 0.001), indicating that uterus dimensions do not drive their increased litter size. These findings provide new insights into the molecular basis of high fertility and could serve as a foundation for further studies on genotype-phenotype relationships in reproductive biology.
{"title":"The special phenotypic characteristics of Dummerstorf superfertile mouse lines could depend on the expression levels of IGF-axis genes","authors":"Michela Calanni-Pileri , Joachim M. Weitzel , Andreas Hoeflich , Martina Langhammer , Marten Michaelis","doi":"10.1016/j.repbio.2025.101012","DOIUrl":"10.1016/j.repbio.2025.101012","url":null,"abstract":"<div><div>To date, animal models with reproductive phenotypes are knockout or transgenic and typically exhibit reduced fertility or infertility. This limits research to studying single-gene effects or loss of fertility. By contrast, Dummerstorf high-fertility mouse lines 1 and 2 (FL1 and FL2) are two unique outbred selection models that demonstrate exceptional reproductive performance. After approximately 50 years of selection, both lines have doubled the number of ovulated oocytes per cycle and consequently their litter size (>20 vs ∼11) compared to the unselected mice of the same founder population (Dummerstorf FZTDU, ctrl line). FL1 and FL2 exhibit atypical estrous cycle length and altered levels of hormones, such as insulin and leptin, which are associated with GnRH release and/or increased body fat content. Unlike typical cases where these factors impair fertility, they instead contribute to the FLs’ high reproductive capacity: the increased ovulation rate results from an upgrade in the quality of their oocytes, influenced by different ovarian lipid profile. In the present study, we analyzed the expression of IGF-axis marker genes linked to reproductive performance and FL-specific traits in three tissues. We found that lepr, which plays a critical role in implantation, was upregulated in the FL1 uterus (1.5-fold vs. ctrl, p < 0.05). In FL1 follicles, igf1, IGF-biding proteins (IGFBP2, IGFBP4) and hsf1—which is involved in gametogenesis—were significantly upregulated (1–4-fold vs. ctrl, p < 0.05 for igf1, hsf1 and IGFBP4; p < 0.01 for igfbp2). In FL2, uterine size was reduced relatively to the body weight (∼0.2 % FL2 vs. 0.25 % in ctrl and 0.28 % in FL1, p < 0.001), indicating that uterus dimensions do not drive their increased litter size. These findings provide new insights into the molecular basis of high fertility and could serve as a foundation for further studies on genotype-phenotype relationships in reproductive biology.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101012"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143724790","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}
Breast cancer with Hashimoto’s thyroiditis (BC-HT) presents a unique immuno-thyroid interplay that remains poorly understood. This study investigates the relationships between thyroid function markers (TSH, T3, T4), immune markers (CD33, CD44), and thyroid autoantibodies (Anti-TPO, Anti-Tg) in BC-HT patients and healthy controls. Normality testing confirmed non-parametric data distribution, necessitating Mann-Whitney U tests for group comparisons. BC-HT patients exhibited significantly elevated TSH, CD33, Anti-TPO, and Anti-Tg levels, alongside reduced T3 and T4, compared to controls, indicating thyroid dysfunction. Spearman’s correlation analysis revealed strong negative correlations between TSH and T3/T4 in controls, which were lost in BC-HT, suggesting disruption of normal thyroid feedback mechanisms. Additionally, CD33 and CD44 correlations with thyroid hormones were evident in controls but absent in BC-HT, highlighting altered immune-thyroid interactions. ROC analysis demonstrated high diagnostic performance for TSH, Anti-Tg, and Anti-TPO, with sensitivities exceeding 0.75, whereas CD33 and CD44 showed limited diagnostic utility. These findings suggest a distinct immuno-thyroid dysregulation in BC-HT patients and highlight the potential of thyroid-specific markers for disease stratification. Future research should focus on longitudinal studies and mechanistic investigations to further delineate the role of immune markers in breast cancer pathophysiology within the context of thyroid autoimmunity.
{"title":"Investigating the clinical significance of immune and thyroid biomarkers in women with breast cancer and Hashimoto's thyroiditis","authors":"Israa Khalaf Aneed , Noori Mohammed Luaibi , Sajid Nader Abdulqader","doi":"10.1016/j.repbio.2025.101011","DOIUrl":"10.1016/j.repbio.2025.101011","url":null,"abstract":"<div><div>Breast cancer with Hashimoto’s thyroiditis (BC-HT) presents a unique immuno-thyroid interplay that remains poorly understood. This study investigates the relationships between thyroid function markers (TSH, T3, T4), immune markers (CD33, CD44), and thyroid autoantibodies (Anti-TPO, Anti-Tg) in BC-HT patients and healthy controls. Normality testing confirmed non-parametric data distribution, necessitating Mann-Whitney <em>U</em> tests for group comparisons. BC-HT patients exhibited significantly elevated TSH, CD33, Anti-TPO, and Anti-Tg levels, alongside reduced T3 and T4, compared to controls, indicating thyroid dysfunction. Spearman’s correlation analysis revealed strong negative correlations between TSH and T3/T4 in controls, which were lost in BC-HT, suggesting disruption of normal thyroid feedback mechanisms. Additionally, CD33 and CD44 correlations with thyroid hormones were evident in controls but absent in BC-HT, highlighting altered immune-thyroid interactions. ROC analysis demonstrated high diagnostic performance for TSH, Anti-Tg, and Anti-TPO, with sensitivities exceeding 0.75, whereas CD33 and CD44 showed limited diagnostic utility. These findings suggest a distinct immuno-thyroid dysregulation in BC-HT patients and highlight the potential of thyroid-specific markers for disease stratification. Future research should focus on longitudinal studies and mechanistic investigations to further delineate the role of immune markers in breast cancer pathophysiology within the context of thyroid autoimmunity.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101011"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143820838","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}
Pub Date : 2025-06-01Epub Date: 2025-03-27DOI: 10.1016/j.repbio.2025.101013
Anna Monallysa Silva de Oliveira , Nívia Maria Rocha Brandão , Anailson De Oliveira Maciel , Nágylla Santos De Almeida , Nayonara Santos De Almeida , Thiago Santos Santos , Izakiel Reis Marinho , Francisco Cardoso Figueiredo , Yndyra Nayan Teixeira Carvalho Castelo Branco , José Adalmir Torres de Souza
The objective was to evaluate the effects of different concentrations of hydroxypropylmethylcellulose (HPMC) on the cryopreservation of bovine semen. Semen was collected from seven bulls by electroejaculation and diluted in Tris-egg yolk medium with and without glycerol (G) in the following treatments: C: Tris-Egg Yolk + 6 %G; T1: Tris-Egg Yolk + 3 % G; T2: Tris-Egg Yolk + 0 % G; T3: Tris-Egg Yolk + 0.5 % HPMC; T4: Tris-Egg Yolk + 1.0 % HPMC; T5: Tris-Egg Yolk + 1.5 % HPMC; T6: Tris-Egg Yolk + 5.5 % G + 0.5 % HPMC. Samples were frozen and stored at −196°C. After thawing, sperm quality was assessed using the rapid thermoresistance test (RTRT), sperm morphology, hypoosmotic swelling test, plasma membrane integrity, mitochondrial activity, and in vitro fertilization (IVF) test. RTRT results showed that the control group had better motility and vigor at 0’, 15’, and 30’ compared to the T6 group. The T6 group had better results than other groups supplemented with 0.5 %, 1.0 %, and 1.5 % HPMC. No significant differences were observed at 45’ for RTRT, hypoosmotic swelling, morphology, membrane integrity, mitochondrial activity, or IVF rates. In conclusion, the addition of HPMC reduced sperm motility and vigor but did not negatively affect other cryopreservation parameters.
目的是评价不同浓度羟丙基甲基纤维素（HPMC）对牛精液冷冻保存的影响。采用电射精法采集7头公牛的精液，在添加和不添加甘油(G)的tris -卵黄培养基中稀释，处理如下：C: tris -卵黄+ 6 %G；T1: Tris-Egg yellow + 3 % G；T2: Tris-Egg yellow + 0 % G；T3: tris -蛋黄+ 0.5 % HPMC；T4: tris -蛋黄+ 1.0 % HPMC；T5: tris -蛋黄+ 1.5 % HPMC；T6: tris -蛋黄+ 5.5 % G + 0.5 % HPMC。样品冷冻保存于- 196°C。解冻后，通过快速耐热性试验（RTRT）、精子形态、低渗肿胀试验、质膜完整性、线粒体活性和体外受精（IVF）试验评估精子质量。RTRT结果显示，与T6组相比，对照组在0 ‘,15 ’和30 '时具有更好的运动性和活力。T6组效果优于添加0.5 %、1.0 %、1.5 % HPMC组。在45 '时，RTRT、低渗肿胀、形态学、膜完整性、线粒体活性或体外受精率均无显著差异。总之，HPMC的加入降低了精子活力和活力，但对其他低温保存参数没有负面影响。
{"title":"Evaluation of the quality of bovine semen subjected to cryopreservation with hydroxypropylmethylcellulose","authors":"Anna Monallysa Silva de Oliveira , Nívia Maria Rocha Brandão , Anailson De Oliveira Maciel , Nágylla Santos De Almeida , Nayonara Santos De Almeida , Thiago Santos Santos , Izakiel Reis Marinho , Francisco Cardoso Figueiredo , Yndyra Nayan Teixeira Carvalho Castelo Branco , José Adalmir Torres de Souza","doi":"10.1016/j.repbio.2025.101013","DOIUrl":"10.1016/j.repbio.2025.101013","url":null,"abstract":"<div><div>The objective was to evaluate the effects of different concentrations of hydroxypropylmethylcellulose (HPMC) on the cryopreservation of bovine semen. Semen was collected from seven bulls by electroejaculation and diluted in Tris-egg yolk medium with and without glycerol (G) in the following treatments: C: Tris-Egg Yolk + 6 %G; T1: Tris-Egg Yolk + 3 % G; T2: Tris-Egg Yolk + 0 % G; T3: Tris-Egg Yolk + 0.5 % HPMC; T4: Tris-Egg Yolk + 1.0 % HPMC; T5: Tris-Egg Yolk + 1.5 % HPMC; T6: Tris-Egg Yolk + 5.5 % G + 0.5 % HPMC. Samples were frozen and stored at −196°C. After thawing, sperm quality was assessed using the rapid thermoresistance test (RTRT), sperm morphology, hypoosmotic swelling test, plasma membrane integrity, mitochondrial activity, and in vitro fertilization (IVF) test. RTRT results showed that the control group had better motility and vigor at 0’, 15’, and 30’ compared to the T6 group. The T6 group had better results than other groups supplemented with 0.5 %, 1.0 %, and 1.5 % HPMC. No significant differences were observed at 45’ for RTRT, hypoosmotic swelling, morphology, membrane integrity, mitochondrial activity, or IVF rates. In conclusion, the addition of HPMC reduced sperm motility and vigor but did not negatively affect other cryopreservation parameters.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101013"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143704779","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}
Pub Date : 2025-06-01Epub Date: 2025-04-19DOI: 10.1016/j.repbio.2025.101020
Shitao Dong, Youbin Liu, Zhimin Yang
Polycystic ovary syndrome (PCOS) poses a significant threat to women's fertility and quality of life. Studies have found a close association between the environmental contaminant tributyltin (TBT) and the occurrence of PCOS. The main objective of this study was to investigate the specific mechanisms by which TBT adversely affects the growth of ovarian granulosa cells. Cell viability, cycle, proliferation, and apoptosis were measured by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT), 5-ethynyl-2’-deoxyuridine (EdU), and flow cytometry. Simultaneously, lactate dehydrogenase (LDH) leakage and Caspase-3 activity were measured by the corresponding kits. Besides, western blot was used to analyze the protein levels of cyclin-dependent kinase inhibitor 1 C (CDKN1C) and the transcription factor Yin Yang 1 (YY1). TBT severely impaired the viability, cell cycle, and proliferation capacity of granulosa cells, and induced their apoptosis. Silencing CDKN1C and YY1 alleviated the damage caused by TBT to the cells, but these repair effects were weakened by CDKN1C overexpressed. By inhibiting the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway, TBT upregulated the YY1-mediated CDNK1C, and further exacerbated the damage to granulosa cells. This study revealed the mechanism that TBT induced the loss of ovarian granulosa cells in PCOS patients by upregulating YY1-mediated CDKN1C expression, which provided new ideas and targets for the pathogenesis and treatment of PCOS.
多囊卵巢综合征（PCOS）严重威胁妇女的生育能力和生活质量。研究发现环境污染物三丁基锡（TBT）与多囊卵巢综合征的发生密切相关。本研究的主要目的是探讨TBT对卵巢颗粒细胞生长不利的具体机制。采用3-（4,5 -二甲基-2-噻唑基）- 2,5 -二苯基-2- h -溴化四唑（MTT）、5-乙基-2 ' -脱氧尿苷（EdU）和流式细胞术检测细胞活力、周期、增殖和凋亡。同时用相应的试剂盒检测乳酸脱氢酶（LDH）渗漏量和Caspase-3活性。western blot检测细胞周期蛋白依赖性激酶抑制剂1 C （CDKN1C）和转录因子阴阳1 （YY1）蛋白水平。TBT严重损害颗粒细胞的活力、细胞周期和增殖能力，诱导颗粒细胞凋亡。沉默CDKN1C和YY1可减轻TBT对细胞的损伤，但这些修复作用因CDKN1C过表达而减弱。TBT通过抑制磷脂酰肌醇3-激酶/蛋白激酶B （PI3K/AKT）信号通路，上调yy1介导的CDNK1C，进一步加重对颗粒细胞的损伤。本研究揭示了TBT通过上调yy1介导的CDKN1C表达诱导PCOS患者卵巢颗粒细胞丢失的机制，为PCOS的发病机制和治疗提供了新的思路和靶点。
{"title":"Tributyltin affects the growth of ovarian granulosa cells in polycystic ovary syndrome by upregulating YY1-mediated CDKN1C via the PI3K/AKT pathway","authors":"Shitao Dong, Youbin Liu, Zhimin Yang","doi":"10.1016/j.repbio.2025.101020","DOIUrl":"10.1016/j.repbio.2025.101020","url":null,"abstract":"<div><div>Polycystic ovary syndrome (PCOS) poses a significant threat to women's fertility and quality of life. Studies have found a close association between the environmental contaminant tributyltin (TBT) and the occurrence of PCOS. The main objective of this study was to investigate the specific mechanisms by which TBT adversely affects the growth of ovarian granulosa cells. Cell viability, cycle, proliferation, and apoptosis were measured by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT), 5-ethynyl-2’-deoxyuridine (EdU), and flow cytometry. Simultaneously, lactate dehydrogenase (LDH) leakage and Caspase-3 activity were measured by the corresponding kits. Besides, western blot was used to analyze the protein levels of cyclin-dependent kinase inhibitor 1 C (CDKN1C) and the transcription factor Yin Yang 1 (YY1). TBT severely impaired the viability, cell cycle, and proliferation capacity of granulosa cells, and induced their apoptosis. Silencing CDKN1C and YY1 alleviated the damage caused by TBT to the cells, but these repair effects were weakened by CDKN1C overexpressed. By inhibiting the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway, TBT upregulated the YY1-mediated CDNK1C, and further exacerbated the damage to granulosa cells. This study revealed the mechanism that TBT induced the loss of ovarian granulosa cells in PCOS patients by upregulating YY1-mediated CDKN1C expression, which provided new ideas and targets for the pathogenesis and treatment of PCOS.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101020"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848160","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}
Relative copy number of mitochondria was estimated for the potential association with the expression of mitochondrial coded stress related genes in bubaline spermatozoa. Semen samples were collected from buffalo bulls under two extreme temperature-humidity index conditions: hot summer and winter. Based on the semen quality, the bulls were categorized: exhibiting poor semen quality during hot summer as "seasonally affected," while those maintaining good semen quality throughout the year, as "seasonally not affected". The average mitochondrial copy numbers were lower during hot summer (15.42 ± 1.2368) compared to winter (17.29 ± 0.72) in both the groups. Furthermore, within the hot summer period, bulls classified as seasonally affected exhibited significantly lower mitochondrial copy numbers (12.86 ± 1.343) than their seasonally unaffected counterpart (17.97 ± 1.34). These results suggest a potential role of mitochondria in influencing semen quality, particularly in response to impaired scrotal thermoregulation during the summer season. Although the fold change in apoptotic genes (BCL2, MCL1, CASP3, and BAK) and oxidative panel genes (CAT, SOD, GPx, ATF4, and FOXO-3), did not differ significantly across the groups, differences were observed between the seasons. Further, to understand the role of copy number in apoptosis and ROS scavenging across the seasons and the groups, the generalized mixed model was employed. The results conveyed a significant negative interaction of copy number with the expression of CAT gene and significant positive interaction of copy number with the apoptotic gene panel. Our findings underscore the significant role of mitochondrial copy numbers in domestic buffalo spermatozoa in managing the challenges of thermoregulation posed by harsh tropical conditions.
{"title":"Evaluation of mitochondrial copy number and gene expression changes in the spermatozoa of buffalo bulls under heat stress","authors":"Meenakshi Chitkara , Harsimran Kaur , Rashi Vasisth , Karpenahalli Ranganatha Sriranga , Ankita Gurao , Karan Mahar , Mahesh Shivanand Dige , R.A.K. Aggarwal , Manishi Mukesh , Pradeep Kumar , Pawan Singh , Ranjit Singh Kataria","doi":"10.1016/j.repbio.2025.101014","DOIUrl":"10.1016/j.repbio.2025.101014","url":null,"abstract":"<div><div>Relative copy number of mitochondria was estimated for the potential association with the expression of mitochondrial coded stress related genes in bubaline spermatozoa. Semen samples were collected from buffalo bulls under two extreme temperature-humidity index conditions: hot summer and winter. Based on the semen quality, the bulls were categorized: exhibiting poor semen quality during hot summer as \"seasonally affected,\" while those maintaining good semen quality throughout the year, as \"seasonally not affected\". The average mitochondrial copy numbers were lower during hot summer (15.42 ± 1.2368) compared to winter (17.29 ± 0.72) in both the groups. Furthermore, within the hot summer period, bulls classified as seasonally affected exhibited significantly lower mitochondrial copy numbers (12.86 ± 1.343) than their seasonally unaffected counterpart (17.97 ± 1.34). These results suggest a potential role of mitochondria in influencing semen quality, particularly in response to impaired scrotal thermoregulation during the summer season. Although the fold change in apoptotic genes (<em>BCL2</em>, <em>MCL1</em>, <em>CASP3</em>, and <em>BAK</em>) and oxidative panel genes (<em>CAT</em>, <em>SOD</em>, <em>GPx</em>, <em>ATF4</em>, and <em>FOXO-3</em>), did not differ significantly across the groups, differences were observed between the seasons. Further, to understand the role of copy number in apoptosis and ROS scavenging across the seasons and the groups, the generalized mixed model was employed. The results conveyed a significant negative interaction of copy number with the expression of <em>CAT</em> gene and significant positive interaction of copy number with the apoptotic gene panel. Our findings underscore the significant role of mitochondrial copy numbers in domestic buffalo spermatozoa in managing the challenges of thermoregulation posed by harsh tropical conditions.</div></div>","PeriodicalId":21018,"journal":{"name":"Reproductive biology","volume":"25 2","pages":"Article 101014"},"PeriodicalIF":2.5,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143747099","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}
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