Reproductive biology最新文献

Therapeutic potential of diosgenin against methotrexate-induced testicular damage in the rat 地奥塞宁对甲氨蝶呤引起的大鼠睾丸损伤的治疗潜力

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-04 DOI: 10.1016/j.repbio.2024.100966

Fatemeh Taleahmad , Mohsen Khalili , Narges Haddadzadeh-Niri , Ensyie Joneidi , Sara Taleahmad , Mehrdad Roghani

This study evaluated diosgenin effects on methotrexate-induced testicular injury in the rats. A single dose of methotrexate (MTX) (20 mg/kg, i.p) was administered, followed by two weeks of diosgenin treatment via gavage starting one day before methotrexate injection. Testicular damage was evaluated through histological examination of seminiferous tubules, as well as analysis of serum testosterone level, oxidative stress and inflammation biomarkers, and antioxidant levels. The results of this study showed that in the MTX-exposed group, oxidative stress indices of malondialdehyde (MDA), reactive oxygen species (ROS), nitrite and indices of inflammation consisting of tumor necrosis factor α (TNFα), and interleukin 6 (IL-6) have a significant increase compared to the control group. Additionally, reductions were observed in antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). In addition, testosterone level decreased and signs of testicular damage were observed in the MTX group. Conversely, in the group treated with diosgenin alongside MTX at a dosage of 50 mg/kg, there was a significant decrease in oxidative stress markers (MDA, ROS, nitrite) and inflammatory markers (TNFα and IL-6). Moreover, there was a significant increase in the levels of antioxidant enzymes (SOD, CAT, and GSH). Diosgenin appears to have the potential to protect testicular tissue from damage caused by the toxic effects of MTX through the reduction of oxidative stress and inflammation.

本研究评估了薯蓣皂苷对甲氨蝶呤引起的大鼠睾丸损伤的影响。研究人员给大鼠注射了单剂量甲氨蝶呤（MTX）（20 毫克/千克，静脉注射），然后从注射甲氨蝶呤的前一天开始灌胃服用地奥司宁两周。通过对生精小管进行组织学检查，以及分析血清睾酮水平、氧化应激和炎症生物标志物以及抗氧化剂水平，对睾丸损伤进行了评估。研究结果表明，与对照组相比，MTX 暴露组的丙二醛（MDA）、活性氧（ROS）、亚硝酸盐等氧化应激指标以及肿瘤坏死因子α（TNFα）和白细胞介素 6（IL-6）等炎症指标均显著增加。此外，还观察到超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽（GSH）等抗氧化酶减少。此外，在 MTX 组还观察到睾酮水平下降和睾丸损伤的迹象。相反，在使用地奥司明和 MTX（剂量为 50 毫克/千克）的组中，氧化应激指标（MDA、ROS、亚硝酸盐）和炎症指标（TNFα 和 IL-6）显著下降。此外，抗氧化酶（SOD、CAT 和 GSH）的水平也有明显提高。薯蓣皂苷似乎有可能通过减少氧化应激和炎症来保护睾丸组织免受 MTX 毒性作用的损害。

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引用次数: 0

Transcriptome analysis of human spermatozoa with different DNA fragmentation index using RNA sequencing 利用 RNA 测序分析不同 DNA 破碎指数的人类精子转录组

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-11-04 DOI: 10.1016/j.repbio.2024.100964

Kailin Yang , Xue Sun , Qiyuan Zheng , Chen Pan , Siyuan Wang , Qingfang Lu , Changlong Xu , Yangqing Lu

The study is aimed to screen the differential expressed genes (DEGs) related to sperm DNA fragmentation in men and provide reference basis of the sperm selection in assisted reproduction based on DNA fragmentation. We evaluated 60 semen samples from patients with high, medium or low sperm DNA fragmentation index (DFI). Using multicolor flow cytometry, we measured the content of reactive oxygen species (ROS), phosphatidylserine (PS) externalization and mitochondrial membrane potential (MMP) in these sperm samples. The results revealed that the more ROS content and PS externalization were detected in the sperm with higher DFI, but there was lower MMP level in the high DFI sperm. Next, we conducted RNA sequencing (RNA-seq) on 3 groups of sperm samples with high, medium and low DFI. Then, Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were performed on DEGs. Furthermore, we utilized qRT-PCR to validated the significantly DEGs from the RNA-seq assay. The transcriptome results showed a total of 5334 DEGs were found in the sperm sample with high, medium and low DFI. According to GO and KEGG analysis 421 down-regulated genes in the high DFI group were related to oxidative stress and spermatogenesis. Thirteen novel genes were also identified that most likely were involved in sperm DNA fragmentation, which were further validated by qRT-PCR. In conclusion, our study suggested that the sperms with highly fragmented DNA were accompanied by down-regulation of a series of genes related to antioxidant and spermatogenesis.

本研究旨在筛选与男性精子DNA碎片相关的差异表达基因（DEGs），为辅助生殖中根据DNA碎片选择精子提供参考依据。我们评估了60份精子DNA碎片指数（DFI）为高、中、低的患者精液样本。我们使用多色流式细胞术测量了这些精子样本中活性氧（ROS）、磷脂酰丝氨酸（PS）外化和线粒体膜电位（MMP）的含量。结果显示，DFI较高的精子中ROS含量和磷脂酰丝氨酸（PS）外化程度较高，但DFI较高的精子中线粒体膜电位（MMP）水平较低。接着，我们对高、中、低DFI的三组精子样本进行了RNA测序（RNA-seq）。然后，我们对 DEGs 进行了基因本体（GO）富集分析和京都基因组百科全书（KEGG）通路分析。此外，我们还利用 qRT-PCR 验证了 RNA-seq 分析中显著的 DEGs。转录组结果显示，高、中、低DFI精子样本中共发现了5334个DEGs。根据 GO 和 KEGG 分析，高 DFI 组中有 421 个下调基因与氧化应激和精子发生有关。研究还发现了 13 个很可能与精子 DNA 断裂有关的新基因，并通过 qRT-PCR 对其进行了进一步验证。总之，我们的研究表明，DNA高度破碎的精子伴随着一系列与抗氧化和精子发生有关的基因的下调。

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引用次数: 0

From genome to epigenome: Who is a predominant player in the molecular hallmarks determining epigenetic mechanisms underlying ontogenesis? 从基因组到表观基因组：谁是决定本体发生的表观遗传机制的分子标志的主要参与者？

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-28 DOI: 10.1016/j.repbio.2024.100965

Marcin Samiec , Monika Trzcińska

Genetic factors are one of the basic determinants affecting ontogenesis in mammals. Nevertheless, on the one hand, epigenetic factors have been found to exert the preponderant and insightful impact on the intracellular mechanistic networks related to not only initiation and suppression, but also up- and downregulation of gene expression in all the phases of ontogenetic development in a variety of mammalian species. On the other hand, impairments in the epigenetic mechanisms underlying reprogramming of transcriptional activity of genes (termed epimutations) not only give rise to a broad spectrum of acute and chronic developmental abnormalities in mammalian embryos, foetuses and neonates, but also contribute to premature/expedited senescence or neoplastic transformation of cells and even neurodegenerative and mental disorders. The current article is focused on the unveiling the present knowledge aimed at the identification, classification and characterization of epigenetic agents as well as multifaceted interpretation of current and coming trends targeted at recognizing the epigenetic background of proper ontogenesis in mammals. Moreover, the next objective of this paper is to unravel the mechanistic insights into a wide array of disturbances leading to molecular imbalance taking place during epigenetic reprogramming of genomic DNA. The above-indicated imbalance seems to play a predominant role in the initiation and progression of anatomo-, histo-, and physiopathological processes throughout ontogenetic development. Conclusively, different modalities of epigenetically assisted therapeutic procedures that have been exemplified in the current article, might be the powerful and promiseful tools reliable and feasible in the medical treatments of several diseases triggered by dysfunctions in the epigenetic landscapes, e.g., myelodysplastic syndromes or epilepsy.

遗传因素是影响哺乳动物本体发育的基本决定因素之一。然而，一方面，在多种哺乳动物的本体发育过程中，人们发现表观遗传因素对细胞内机制网络产生了重大而深刻的影响，这些网络不仅涉及基因表达的启动和抑制，还涉及基因表达的上调和下调。另一方面，作为基因转录活性重编程基础的表观遗传学机制（称为表突变）的损伤不仅会导致哺乳动物胚胎、胎儿和新生儿出现各种急性和慢性发育异常，还会导致细胞过早/过快衰老或肿瘤性转化，甚至神经退行性疾病和精神疾病。本文的重点是揭示表观遗传因子的识别、分类和特征描述方面的现有知识，以及对当前和未来趋势的多方面解读，旨在认识哺乳动物正常本体发育的表观遗传背景。此外，本文的下一个目标是揭示在基因组 DNA 表观遗传重编程过程中导致分子失衡的一系列干扰的机理。上述失衡似乎在整个本体发育过程中的解剖学、组织学和生理病理学过程的启动和进展中起着主导作用。总之，本文举例说明的不同模式的表观遗传辅助治疗程序，可能是治疗由表观遗传景观功能障碍引发的多种疾病（如骨髓增生异常综合征或癫痫）的可靠而可行的强大工具。

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引用次数: 0

The alteration in myometrial mRNA transcription of the regulatory genes of DNA methylation in mare with endometrosis 子宫内膜异位症母马子宫肌层 mRNA 转录 DNA 甲基化调控基因的改变。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-22 DOI: 10.1016/j.repbio.2024.100962

Beatriz Celeiro e Silva , Ewa Monika Drzewiecka , Katarzyna Piotrowska-Tomala , Joana Alpoim-Moreira , Agnieszka Sadowska , Magdalena Karolina Kowalik , Jorge Pimenta , Maria Rosa Rebordão , Graça Ferreira-Dias , Dariusz Skarzynski , Anna Szóstek-Mioduchowska

A reduction in myometrial contractile activity can lead to inadequate cleaning of the uterine lumen, resulting in persistent endometritis and potentially endometrosis in mares. Oxytocin (OXT) is a key hormonal regulator of myometrial contraction. While epigenetic regulation of myometrial gene expression has been studied in humans, there is limited information on the expression of DNA methyltransferases (DNMTs) and ten-eleven translocation enzymes (TETs) in the myometrium of mares. This study aimed to evaluate the mRNA transcription of these enzymes and the potential role of DNA methylation in the expression of the OXT receptor (OXTR) gene in the myometrium of mares with endometrosis. Myometrial samples were collected post-mortem during the mid-luteal (n = 23) and follicular (n = 20) phases of the estrous cycle and assessed according to Kenney and Doig endometrial category (I, IIA, IIB, III). mRNA transcription of OXTR, DNMT1, -3A, -3B and TET1, -2, -3 were determined using qPCR. DNA methylation analysis at CpG islands of OXTR exons 1 and 2 was performed using bisulfite pyrosequencing. Myometrial OXTR mRNA transcription and DNA methylation in its promoter region showed no significant differences between categories, although increased methylation was observed at CpG island position 6 in exon 2. DNMT1, TET2, and TET3 mRNA transcription was altered in the equine myometrium depending on the phase of the estrous cycle and the severity of endometrosis (P < 0.05). These findings indicate that DNMTs and TETs were expressed in myometrium in a manner specific to the severity of endometrosis and phases of the estrous cycle, suggesting a potential regulatory role in DNA methylation of myometrial gene expression.

子宫肌收缩活动减弱会导致子宫腔清洁不足，造成持续性子宫内膜炎，甚至可能导致母马子宫内膜坏死。催产素（OXT）是子宫肌收缩的关键激素调节因子。虽然对人类子宫肌层基因表达的表观遗传调控已有研究，但有关母马子宫肌层 DNA 甲基转移酶（DNMTs）和十-十一转位酶（TETs）表达的信息却很有限。本研究旨在评估这些酶的 mRNA 转录情况，以及 DNA 甲基化在子宫内膜异位症母马子宫肌层 OXT 受体（OXTR）基因表达中的潜在作用。在发情周期的黄体中期（n = 23）和卵泡期（n = 20）收集子宫肌层样本，并根据 Kenney 和 Doig 的子宫内膜类别（I、IIA、IIB、III）进行评估，使用 qPCR 测定 OXTR、DNMT1、-3A、-3B 和 TET1、-2、-3 的 mRNA 转录。使用亚硫酸氢盐热测序法对 OXTR 外显子 1 和 2 的 CpG 岛进行了 DNA 甲基化分析。子宫肌层 OXTR mRNA 的转录及其启动子区域的 DNA 甲基化在不同类别之间没有明显差异，但在外显子 2 的 CpG 岛第 6 位观察到甲基化增加。马子宫肌层中 DNMT1、TET2 和 TET3 mRNA 的转录随发情周期的阶段和子宫内膜坏死的严重程度而改变（P＜0.05）。

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引用次数: 0

Negative effects of ketoprofen and meloxicam on distal cauda epidydimal duct contractions, testosterone levels, and sperm count in rats 酮洛芬和美洛昔康对大鼠远端尾状腱收缩、睾酮水平和精子数量的负面影响

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-21 DOI: 10.1016/j.repbio.2024.100963

Ramon Tadeu Galvão Alves Rodrigues , Vitória Barros Marques , Maria Santana da Silva , Luana Talinne da Costa Gomes , Maele Oliveira de Sena , Bruna da Silva Figueiredo , Jonas Ivan Nobre Oliveira , Elaine Cristina Gavioli , Danilo José Ayres de Menezes , Edilson Dantas da Silva Junior

Ketoprofen and meloxicam, non-steroidal anti-inflammatory drugs widely used in clinical practice, lack comprehensive investigation regarding their impact on male reproductive health, particularly on epididymal duct contractions and sperm parameters. Therefore, this study investigated the negative effects of ketoprofen or meloxicam on the contractions of the epididymal duct, sperm parameters, and serum testosterone levels in rats. Firstly, we assessed the in vitro effects of ketoprofen or meloxicam (1–100 μM) on the contractions of the epididymal duct elicited by noradrenaline. Rats were also orally treated with 5 mg/kg ketoprofen or 1 mg/kg meloxicam for 15 days following evaluation of epididymal duct contractions, sperm parameters, and serum testosterone levels. In vitro exposure to meloxicam (100 μM), but not ketoprofen, significantly reduced the maximum effect of noradrenaline in epididymal duct. Moreover, in vivo administration of ketoprofen and meloxicam decreased testosterone levels, sperm production, and sperm count in the caput/corpus region of the rat epididymis. Conversely, the sperm count in the cauda epididymis remained unchanged in animals treated with both ketoprofen and meloxicam. Meloxicam, but not ketoprofen, caused a delay in sperm transit time in the cauda region of the epididymis. In vivo treatment with both ketoprofen or meloxicam hindered the noradrenaline-induced contractions in the epididymal duct. In conclusion, ketoprofen and meloxicam can modify sperm parameters by decreasing testosterone levels and the contractions of the epididymal duct isolated from the distal cauda region of the rat epididymis.

酮洛芬和美洛昔康是广泛应用于临床的非甾体抗炎药，但它们对男性生殖健康的影响，尤其是对附睾管收缩和精子参数的影响，还缺乏全面的研究。因此，本研究探讨了酮洛芬或美洛昔康对大鼠附睾管收缩、精子参数和血清睾酮水平的负面影响。首先，我们在体外评估了酮洛芬或美洛昔康（1-100 μM）对去甲肾上腺素引起的附睾管收缩的影响。在对大鼠的附睾管收缩、精子参数和血清睾酮水平进行评估后，还对大鼠口服 5 毫克/千克酮洛芬或 1 毫克/千克美洛昔康，为期 15 天。体外暴露于美洛昔康（100 μM）而非酮洛芬可显著降低去甲肾上腺素在附睾管中的最大作用。此外，体内注射酮洛芬和美洛昔康可降低大鼠附睾睾帽/睾丸区域的睾酮水平、精子产量和精子数量。相反，接受酮洛芬和美洛昔康治疗的动物附睾尾部的精子数量保持不变。美洛昔康（而非酮洛芬）会延迟精子在附睾尾部的转运时间。在体内使用酮洛芬或美洛昔康会阻碍去甲肾上腺素引起的附睾管收缩。总之，酮洛芬和美洛昔康可以通过降低睾酮水平和从大鼠附睾远端尾区分离出的附睾管的收缩来改变精子参数。

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引用次数: 0

Partial biochemical and immunological characterization of an isolated 52.1 kDa pregnancy-associated glycoprotein charge variant 分离出的 52.1 kDa 妊娠相关糖蛋白电荷变体的部分生物化学和免疫学特征

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-18 DOI: 10.1016/j.repbio.2024.100960

Gerardo Perera-Marín , Giovanna León-Legaspi , Everardo González-Padilla , Clara Murcia , Rogelio Alonso-Morales , Silvia Ivonne Mora Herrera , Griselda Valdez-Magaña

Pregnancy-associated glycoproteins (PAGs) are synthesized in the placental cells of ruminants and are detectable in blood, milk, and urine. Many of these proteins have been obtained and characterized from placental extracts by precipitation with 80 % ammonium sulfate. The possibility of purifying PAGs by precipitation with other concentrations of ammonium sulfate remains unexplored. We aimed to study PAG proteins obtained from extracts of ovine placenta at 100 days of gestation through precipitation with 40 % ammonium sulfate (Extract 40). The main protein complex (130 kDa) was obtained after Extract 40 precipitation. Under reducing SDS-PAGE conditions, the 130 kDa complex dissociated in two PAG proteins with apparent molecular weights of 52.1 kDa and 26.1 kDa. The 130 kDa protein appeared to be a molecular complex consisting of two copies of the 52.1 kDa protein linked to one copy of the 26.1 kDa protein, presumably by disulfide bonds. Furthermore, the 52.1 kDa protein consisted of at least three isoforms with distinct isoelectric points. Amino acid microsequencing of the 52.1 kDa protein revealed a chimeric structure containing amino acid sequences of PAG1, PAG4, PAG6, and PAG1-like proteins. This procedure recovered a novel 130 kDa protein complex composed of 26.1 kDa and two 52.1 kDa PAGs. To the best of our knowledge, this has not been previously reported as heterologous polymeric molecules.

妊娠相关糖蛋白（PAG）在反刍动物的胎盘细胞中合成，可在血液、乳汁和尿液中检测到。其中许多蛋白质都是通过 80% 硫酸铵沉淀法从胎盘提取物中获得并表征的。用其他浓度的硫酸铵沉淀纯化 PAG 的可能性仍有待探索。我们的目的是研究用 40% 硫酸铵（提取物 40）沉淀从妊娠 100 天的绵羊胎盘提取物中获得的 PAG 蛋白。提取物 40 沉淀后得到了主要的蛋白质复合物（130 kDa）。在还原 SDS-PAGE 条件下，130 kDa 复合物解离成两个 PAG 蛋白，表观分子量分别为 52.1 kDa 和 26.1 kDa。130 kDa 蛋白似乎是由两个 52.1 kDa 蛋白拷贝与一个 26.1 kDa 蛋白拷贝（可能是通过二硫键连接）组成的分子复合物。此外，52.1 kDa 蛋白至少由三种等电点不同的异构体组成。对 52.1 kDa 蛋白的氨基酸微序列分析表明，它是一种嵌合结构，包含 PAG1、PAG4、PAG6 和 PAG1-like 蛋白的氨基酸序列。这一过程发现了一个由 26.1 kDa 和两个 52.1 kDa PAG 组成的新型 130 kDa 蛋白复合物。据我们所知，以前从未报道过这种异源聚合分子。

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引用次数: 0

Effect and mechanisms of cyclophosphamide-induced ovarian toxicity on the quality of primordial follicles with respect to age at treatment initiation 环磷酰胺引起的卵巢毒性对原始卵泡质量的影响和机制与开始治疗时的年龄有关

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-14 DOI: 10.1016/j.repbio.2024.100959

Motoki Takenaka , Hinako M. Takase , Noriko N. Suzuki , Chiemi Saigo , Tamotsu Takeuchi , Tatsuro Furui

Chemotherapy-induced ovarian toxicity in patients with cancer significantly affects future fertility depending on the age of initiation of treatment. However, the mechanisms underlying the age-related depletion of the ovarian reserve are not well understood. We investigated the effects of chemotherapy on pre- and postpubertal ovarian reserves in a mouse model. Juvenile (3-week-old) and adult (8-week-old) mice were injected with vehicle or cyclophosphamide (CPA;100 mg/kg). We assessed the short-term effects at 24 h and 72 h after injection and the long-term effects at 10 and 12 weeks of age by counting the follicles. The number of primordial follicles in the juvenile group was significantly reduced by CPA treatment compared with that in the adult group. To elucidate the mechanisms of this depletion, we performed immunostaining for γH2AX, cleaved PARP1, and FOXO3 at 24 h post-treatment. CPA-treated juvenile mice had a significantly higher proportion of γH2AX-positive primordial follicles, indicating double-strand DNA breaks. By contrast, 4-hydroperoxy CPA, an activated analog of CPA, induced γH2AX-positive primordial follicles in both groups in vitro, suggesting age-dependent differences in humoral ovarian microenvironment. Moreover, the level of cleaved PARP1 was specifically elevated in CPA-treated juvenile mice. However, primordial follicle activation was unaffected in the CPA-treated groups, as assessed by FOXO3 translocation. In conclusion, our findings suggest that ovaries in juveniles are more susceptible to DNA damage and subsequent apoptosis, leading to a higher rate of primordial follicle depletion. Therefore, it is crucial to recognize that cancer treatment, especially in children, can exert a substantial influence on future fertility.

癌症患者因化疗引起的卵巢毒性会严重影响未来的生育能力，这取决于开始治疗的年龄。然而，与年龄相关的卵巢储备耗竭的机制尚不十分清楚。我们在小鼠模型中研究了化疗对青春期前和青春期后卵巢储备的影响。给幼年（3周大）和成年（8周大）小鼠注射药物或环磷酰胺（CPA；100 mg/kg）。我们在注射后 24 小时和 72 小时评估了短期效应，并在 10 周龄和 12 周龄时通过计数卵泡评估了长期效应。与成年组相比，幼年组的原始卵泡数量在CPA治疗后明显减少。为了阐明这种减少的机制，我们在处理后24小时对γH2AX、裂解的PARP1和FOXO3进行了免疫染色。经 CPA 处理的幼鼠原始卵泡中 γH2AX 阳性的比例明显更高，这表明存在双链 DNA 断裂。相比之下，CPA的活化类似物4-hydroperoxy CPA在体外诱导γH2AX阳性原始卵泡的比例在两组小鼠中都很高，这表明体液卵巢微环境的差异与年龄有关。此外，在 CPA 处理的幼年小鼠中，PARP1 的裂解水平特异性升高。然而，根据FOXO3转位评估，CPA处理组的原始卵泡活化不受影响。总之，我们的研究结果表明，幼鼠的卵巢更容易受到 DNA 损伤和随后的细胞凋亡，导致原始卵泡耗竭率更高。因此，认识到癌症治疗（尤其是儿童癌症治疗）会对未来生育能力产生重大影响至关重要。

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引用次数: 0

Localization and functional analysis of miR-92a-3p regulating Ino80d in mouse testis 小鼠睾丸中调控 Ino80d 的 miR-92a-3p 的定位和功能分析

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-13 DOI: 10.1016/j.repbio.2024.100961

Lvjing Luo , Lishuang Sun , Shu Li , Huiting Liu , Shi Huang , Yinyin Mo , Genliang Li

Testicular development and spermatogenesis in mice involve complex and dynamic gene regulation and chromatin remodelling. In this study, Real-time fluorescence quantitative PCR (RT-qPCR), Western Blot (WB), Immunofluorescence (IF), transfection and other techniques were used to analyse the expression of Ino80d mRNA and its encoded proteins in mouse testicular tissue and mouse spermatogonial cells, and to further analyse the possible target-regulatory relationship and function of miR-92a-3p and Ino80d. We found that Ino80d mRNA and protein expression was up-regulated in adult mouse testis tissue relative to juvenile mouse testis tissue, whereas miR-92a-3p expression was down-regulated in adult mouse testis tissue. Immunofluorescence results showed that the Ino80d protein was mainly localized in the nucleus of male germ cells. Ino80d protein expression is higher in spermatogonia, spermatid and lower in primary spermatocytes, secondary spermatocytes and sperm. There is a decreasing trend in development from spermatogonia to secondary spermatocytes. The transfection results showed that the expression levels of Ino80d mRNA and protein were down-regulated after overexpression of miR-92a-3p in mouse spermatogonia. Increased miR-92a-3p may be a key factor in inhibiting the expression of Ino80d mRNA and proteins in the miR-92a-3p mimics group of mouse spermatogonial cells, whereas differential expression may be a result of the negative regulation of miR-92a-3p, which regulates testicular development and spermatogenesis in mice.

小鼠的睾丸发育和精子发生涉及复杂而动态的基因调控和染色质重塑。本研究采用实时荧光定量 PCR（RT-qPCR）、Western Blot（WB）、免疫荧光（IF）、转染等技术分析了 Ino80d mRNA 及其编码蛋白在小鼠睾丸组织和小鼠精原细胞中的表达，并进一步分析了 miR-92a-3p 与 Ino80d 可能的靶调控关系和功能。我们发现，与幼鼠睾丸组织相比，成年小鼠睾丸组织中 Ino80d mRNA 和蛋白表达上调，而 miR-92a-3p 在成年小鼠睾丸组织中表达下调。免疫荧光结果显示，Ino80d 蛋白主要定位于雄性生殖细胞的细胞核中。Ino80d蛋白在精原细胞和精母细胞中表达较高，而在初级精母细胞、次级精母细胞和精子中表达较低。从精原细胞到次级精母细胞的发育过程中，Ino80d 蛋白表达呈下降趋势。转染结果显示，在小鼠精原细胞中过表达 miR-92a-3p 后，Ino80d mRNA 和蛋白质的表达水平下调。在 miR-92a-3p 模拟组小鼠精原细胞中，miR-92a-3p 的增加可能是抑制 Ino80d mRNA 和蛋白表达的关键因素，而不同的表达可能是 miR-92a-3p 负调控的结果，它调控着小鼠的睾丸发育和精子生成。

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引用次数: 0

Exploring stem cell technology: Pioneering new pathways for female fertility preservation and restoration 探索干细胞技术：开辟女性生育能力保存和恢复的新途径。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-10 DOI: 10.1016/j.repbio.2024.100958

Ningjing Li , Xinrong Du , Yuhong Zhao , Qin Zeng , Changli Han , Dongsheng Xiong , Libing He , Guohui Zhang , Weixin Liu

The fertility of women is crucial for the well-being of individuals and families. However, various factors such as chemotherapy, lifestyle changes, among others, may lead to a decline in female fertility, thus emphasizing the significance of preserving and restoring fertility. Stem cells, with their unique capacity for self-renewal and pluripotent differentiation, have made significant strides in areas such as ovarian tissue cryopreservation, in vitro culture of frozen-thawed ovarian tissue, and construction of ovarian-like organs. This review aims to summarize the latest findings in these fields, highlighting the pivotal role, mechanisms, and future prospects of stem cell technology in preserving and restoring female fertility. Additionally, the importance of interdisciplinary collaboration is underscored, as personalized stem cell therapy regimens tailored through interdisciplinary cooperation between reproductive medicine and stem cell fields hold promise in providing reliable solutions for the preservation and restoration of female fertility.

女性的生育能力对个人和家庭的幸福至关重要。然而，化疗、生活方式的改变等各种因素都可能导致女性生育能力下降，因此，保持和恢复生育能力的意义就显得尤为重要。干细胞具有独特的自我更新和多能分化能力，在卵巢组织冷冻保存、冻融卵巢组织体外培养和卵巢类器官构建等领域取得了重大进展。本综述旨在总结这些领域的最新研究成果，强调干细胞技术在保存和恢复女性生育能力方面的关键作用、机制和未来前景。此外，还强调了跨学科合作的重要性，因为通过生殖医学和干细胞领域的跨学科合作定制的个性化干细胞治疗方案，有望为保存和恢复女性生育能力提供可靠的解决方案。

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引用次数: 0

Exposure to chronic stress impedes seasonal and gonadotropin-induced ovarian recrudescence in the gecko Hemidactylus frenatus 慢性应激会阻碍壁虎半爪蜥季节性和促性腺激素诱导的卵巢再发育。

IF 2.5 3区生物学 Q3 REPRODUCTIVE BIOLOGY

Reproductive biology

Pub Date : 2024-10-07 DOI: 10.1016/j.repbio.2024.100957

Ananya Ganeyan, C.B. Ganesh

The neuroendocrine regulation of the stress-reproductive axis in reptiles is complex due to the diverse reproductive strategies adopted by these animals. Consequently, the underlying mechanisms by which stress can affect the reproductive axis remain opaque in reptiles. In the present study, we examined the effect of stress on the seasonal and FSH-induced ovarian recrudescence during the breeding and non-breeding phases of the cycle in the tropical and subtropical house gecko Hemidactylus frenatus. During the recrudescence phase of the ovarian cycle, exposure of lizards to various stressors (handling, confinement, chasing, and noise) caused a significant increase in the percentage of corticotropin-releasing hormone (CRH) and adrenocorticotropic hormone (ACTH)-immunoreactive (ir) content in the median eminence (ME) and/or pars distalis of the pituitary gland (PD), concomitant with a significant decrease in the release of gonadotropin-releasing hormone (GnRH)-ir content into the ME and PD, and number of oogonia in the germinal bed and absence of the stage IV and V (vitellogenic) follicles in the ovary compared to experimental controls. During the non-breeding phase, treatment of stressed lizards with FSH did not stimulate the development of stage IV and V follicles, in contrast to their appearance in FSH-only-treated lizards. Collectively, these findings suggest that exposure to stressors prevents the seasonal ovarian recrudescence, possibly mediated through the suppression of hypothalamic GnRH release into the ME and PD and/or directly at the level of the ovary.

爬行动物的应激-生殖轴的神经内分泌调控非常复杂，因为这些动物采取的生殖策略多种多样。因此，爬行动物应激影响生殖轴的内在机制仍然不清楚。在本研究中，我们考察了应激对热带和亚热带家壁虎（Hemidactylus frenatus）繁殖和非繁殖周期中季节性和FSH诱导的卵巢复旧的影响。在卵巢周期的再发育阶段，蜥蜴暴露于各种应激源（搬运、禁闭、追逐和噪音）会导致垂体正中突起和/或垂体远端旁的促肾上腺皮质激素释放激素（CRH）和促肾上腺皮质激素（ACTH）免疫反应性（ir）含量的百分比显著增加、同时，与实验对照组相比，促性腺激素释放激素（GnRH）-ir 含量在中脑和垂体上部的释放量、生殖床中卵原细胞的数量以及卵巢中第 IV 期和第 V 期（卵黄形成期）卵泡的缺失均显著减少。在非繁殖期，用FSH处理应激蜥蜴不会刺激IV期和V期卵泡的发育，而只用FSH处理的蜥蜴则没有这种现象。总之，这些研究结果表明，暴露于应激源可防止季节性卵巢复旧，这可能是通过抑制下丘脑GnRH向ME和PD的释放和/或直接在卵巢水平介导的。

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引用次数: 0