Animal Reproduction Science最新文献_第2页

Dihydrotestosterone binding to the androgen receptor inhibits apoptosis in ovine undifferentiated granulosa cells via AMH signaling 双氢睾酮结合雄激素受体通过AMH信号抑制绵羊未分化颗粒细胞凋亡

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2026-01-12 DOI: 10.1016/j.anireprosci.2026.108105

Yufen Zhao , Haijiang Liu , Zhe Mu , Haijun Li

Although dihydrotestosterone (DHT), androgen receptor (AR), and anti-Müllerian hormone (AMH) play key roles in ovarian function, their precise contributions and synergistic roles in mammalian follicular development remain unclear. In this study, we first evaluated whether ovine granulosa cells (GCs) undergo differentiation during vitro culture and then determined methods for inhibiting differentiation. Following the culture of GCs with DHT (10⁻⁷ mol/L) and the AR antagonist enzalutamide (ENZ) (10⁻⁶ mol/L) separately or in combination for 48 h, an apoptosis assay revealed that the interaction between DHT and AR decreases sheep GC apoptosis. DHT significantly increased the AMH concentration through AR signaling. In addition, DHT and AR significantly increased the intracellular AMH level, and the use of rAMH (70 pg/mL), a sheep recombinant AMH, significantly upregulated the expression of the BCL2 mRNA in GCs. In summary, GCs undergo spontaneous differentiation during in vitro culture, and the use of the specific luteinizing hormone receptor (LHR) antagonist BAY-899 to inhibit their differentiation establishes an in vitro culture model that maintains GC characteristics. Thus, the interaction between DHT and AR may be involved in the accumulation of AMH signals, affecting GC apoptosis. We provide new mechanistic insights into the roles of DHT and AR in attenuating sheep GC apoptosis. The finding that potential cross-points between DHT/AR-induced intracellular signals affect GC apoptosis will help elucidate the mechanisms regulating early follicular development.

尽管双氢睾酮（DHT）、雄激素受体（AR）和抗勒氏激素（AMH）在卵巢功能中起着关键作用，但它们在哺乳动物卵泡发育中的确切作用和协同作用尚不清楚。在这项研究中，我们首先评估了绵羊颗粒细胞（GCs）在体外培养过程中是否发生分化，然后确定了抑制分化的方法。DHT（10−7 mol/L）和AR拮抗剂恩扎鲁胺（enzz）（10−6 mol/L）分别或联合培养GCs 48 h后，细胞凋亡实验显示DHT和AR相互作用可减少绵羊GC细胞凋亡。DHT通过AR信号通路显著提高AMH浓度。此外，DHT和AR显著提高了细胞内AMH水平，rAMH(70 pg/mL)，一种绵羊重组AMH，显著上调了GCs中BCL2 mRNA的表达。综上所述，GC在体外培养过程中自发分化，利用特异性黄体生成素受体（LHR）拮抗剂BAY-899抑制其分化，建立了维持GC特征的体外培养模型。因此，DHT与AR的相互作用可能参与了AMH信号的积累，影响了GC的凋亡。我们为DHT和AR在减轻绵羊GC细胞凋亡中的作用提供了新的机制见解。DHT/ ar诱导的细胞内信号之间的潜在交叉点影响GC凋亡的发现将有助于阐明调节早期卵泡发育的机制。

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引用次数: 0

The Pma-PABP4 gene, regulated by Pma-miR-1444, participates in Pinctada fucata gonad development Pma-miR-1444调控的Pma-PABP4基因参与中华绒螯虾性腺发育

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2026-01-11 DOI: 10.1016/j.anireprosci.2026.108106

Ying Guo , Lirong Bai , Pei Wang , Tangrongjun Guo , Ruolin Zhu , Weiwei Chen , Xiuxiu Sang , Xianwei Fan , Dahui Yu

This study cloned and characterized the full-length cDNA of the Pma-PABP4 gene from Pinctada fucata, which is 3123 bp in length and encodes a 641-amino acid protein. The protein exhibited typical RNA-binding protein characteristics, including four RNA recognition motifs (RRMs) and a poly(A)-binding domain. Bioinformatics analyses revealed that Pma-PABP4 is hydrophobic and contains multiple predicted phosphorylation sites. Phylogenetic analysis showed that Pma-PABP4 is highly homologous to PABP4 from other mollusks and shares evolutionary conservation with human PABP1. Additionally, Pma-PABP4 was widely expressed in many tissues in P. fucata with sex-biased expression, where expression levels varied significantly during early embryonic development and specific stages of gonadal development. It was also localized in the gonadal tissues, and RNA interference experiments confirmed that silencing Pma-PABP4 resulted in abnormal gonadal development and reduced gamete production. This study also showed that Pma-miR-1444 directly targets the coding sequence of Pma-PABP4 at the 1371-bp site, negatively regulating its expression. The spatiotemporal expression patterns of Pma-miR-1444 were also closely associated with gametogenesis and embryonic development. Overexpression of Pma-miR-1444 led to gonadal atrophy and a significant reduction in gamete numbers. In conclusion, Pma-miR-1444 and Pma-PABP4 play crucial regulatory roles in the reproductive development and embryogenesis of P. fucata.

本研究克隆并鉴定了fucata pintada Pma-PABP4基因全长cDNA，全长3123 bp，编码641个氨基酸的蛋白。该蛋白具有典型的RNA结合蛋白特征，包括4个RNA识别基序（rrm）和一个聚(a)结合结构域。生物信息学分析显示Pma-PABP4是疏水性的，含有多个可预测的磷酸化位点。系统发育分析表明，Pma-PABP4与其他软体动物的PABP4高度同源，与人类PABP1具有相同的进化保守性。此外，Pma-PABP4在fucata的许多组织中广泛表达，并具有性别偏倚表达，其表达水平在胚胎早期发育和性腺发育的特定阶段发生显著变化。它也定位于性腺组织，RNA干扰实验证实，沉默Pma-PABP4会导致性腺发育异常和配子产生减少。本研究还发现，Pma-miR-1444直接靶向Pma-PABP4编码序列的1371-bp位点，负向调控其表达。Pma-miR-1444的时空表达模式也与配子体发生和胚胎发育密切相关。Pma-miR-1444的过表达导致性腺萎缩和配子数量的显著减少。综上所述，Pma-miR-1444和Pma-PABP4在fucata的生殖发育和胚胎发生中起着至关重要的调节作用。

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引用次数: 0

Equine fetal-sex determination from mid-gestation to term using serum conjugated steroid profiling by liquid chromatography-tandem mass spectrometry 用液相色谱-串联质谱法测定马妊娠中期至足月胎儿性别

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2026-01-03 DOI: 10.1016/j.anireprosci.2026.108098

Joy Ledeck , Thomas Dubrowski , Matthieu Schoumacher , Stéphanie Peeters , Caroline Le Goff , Sophie Egyptien , Stéfan Deleuze , Etienne Cavalier , Jérôme Ponthier

Refined profiling of conjugated estrogens and androgens during equine pregnancy using liquid chromatography-tandem mass spectrometry (LC-MS/MS) could provide accurate fetal-sex determination. Current methods for fetal-sex prediction remain limited by timing, accuracy, and operator expertise. This study investigated sex-specific differences in maternal conjugated steroid profiles to develop a reliable, non-invasive predictive method. Samples were collected from 141 mares of various breeds starting at sixteen weeks of pregnancy. The samples were pooled according to gestational stage, divided into 28-day blocks, and analysed using a validated LC-MS/MS method. Quantified analytes included estrone, estradiol, equilin sulfates, glucuronides, and dehydroepiandrosterone sulfate. Method validation encompassed linearity, trueness, precision, accuracy, uncertainty, quantification limits, recovery, matrix effects, carryover, sensitivity, and stability. The effects of fetal sex, breed, parity, and maternal age on steroid concentrations were investigated. Data were split into a Test group for model development and a Validation group for performance assessment. Fetal sex was the principal factor influencing conjugated estrogen and dehydroepiandrosterone profiles (p = 0.01–0.0001). Female pregnancies exhibited higher classical conjugated steroid concentrations at block 5, whereas male pregnancies showed delayed elevations persisting until term. Equilin derivatives were consistently lower in males. Estrone sulfate (E1S) showed reliable univariate performance for fetal-sex prediction (area under the curve (AUC) 0.710–0.865, blocks 5–10). Multivariate models using all conjugated steroids achieved superior accuracy (AUC 0.747–0.912, blocks 5–11), providing an alternative method to univariate model and ultrasonography. The validated LC-MS/MS method improves understanding of equine feto-placental endocrinology and offers a non-invasive tool for practical fetal sexing.

利用液相色谱-串联质谱法（LC-MS/MS）对马妊娠期间的共轭雌激素和雄激素进行精细分析，可以提供准确的胎儿性别测定。目前的胎儿性别预测方法仍然受到时间、准确性和操作人员专业知识的限制。本研究调查了母体偶联类固醇谱的性别特异性差异，以开发一种可靠的、非侵入性的预测方法。从怀孕16周开始，从141匹不同品种的母马身上收集样本。根据妊娠阶段收集样本，分成28天块，使用经过验证的LC-MS/MS方法进行分析。定量分析物包括雌酮、雌二醇、硫酸equilin、葡萄糖醛酸酯和硫酸脱氢表雄酮。方法验证包括线性、真实度、精密度、准确度、不确定度、定量限、回收率、基质效应、转移、灵敏度和稳定性。研究了胎儿性别、品种、胎次和母亲年龄对类固醇浓度的影响。数据被分成用于模型开发的Test组和用于性能评估的Validation组。胎儿性别是影响共轭雌激素和脱氢表雄酮水平的主要因素（p = 0.01-0.0001）。在阻滞5期，女性妊娠表现出较高的经典偶联类固醇浓度，而男性妊娠表现出延迟升高，持续到足月。Equilin衍生物在雄性中的含量一直较低。硫酸雌酮（E1S）在胎儿性别预测方面表现出可靠的单变量性能（曲线下面积（AUC） 0.710-0.865，区块5-10）。使用所有共轭类固醇的多变量模型获得了更高的准确性（AUC为0.747-0.912，区块5-11），为单变量模型和超声检查提供了另一种方法。经验证的LC-MS/MS方法提高了对马胎儿-胎盘内分泌学的理解，并为实际胎儿性别鉴定提供了一种无创工具。

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引用次数: 0

Fertilization in farmed and wild maraena whitefish (Coregonus maraena, Bloch, 1779) in different activation solutions 养殖和野生maraena白鱼（Coregonus maraena, Bloch, 1779）在不同活化溶液中的施肥。

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2026-01-01 DOI: 10.1016/j.anireprosci.2025.108097

Beata Irena Cejko , Dorota Fopp-Bayat , Roman Kujawa , Agata Anna Cejko

This study compared the reproductive performance of farmed and wild maraena whitefish (Coregonus maraena) and fertilization rates of sperm activated with three activation solutions (AS): Billard solution (BS: 20 mM Tris containing 1 mM CaCl₂, 30 mM glycine, and 154 mM NaCl; pH 9.0; osmolality 330 mOsm·kg⁻¹), saline solution (SS: 20 mM Tris containing 30 mM NaCl; pH 8.5; 80 mOsm·kg⁻¹), and lake water (LW: pH 7.5; 20 mOsm·kg⁻¹). Although no significant differences were observed between farmed and wild fish in basic reproductive traits (egg weight, pseudogonado-somatic index, sperm volume, sperm production), notable contrasts emerged in sperm kinetics and fertilization success. Progressive motility (PRG), curvilinear velocity (VCL), straight-line velocity (VSL), wobble (WOB), and beat cross frequency (BCF) changed significantly depending on the AS used for sperm activation in both farmed and wild whitefish. There were no significant differences in fertilization rate between farmed and wild fish after BS used. However, compared to farmed, in wild fish, fertilization rates were higher with SS and LW, despite lower sperm motility under these conditions. Linear regression analysis revealed significant negative correlations between high sperm velocity and fertilization success in wild males after LW activation, suggesting that excessive motility may deplete sperm energy reserves prematurely. These findings highlight that optimal, rather than maximal, motility may be more critical for successful fertilization in maraena whitefish particularly in wild stocks. However, this hypothesis requires further research.

本研究比较了繁殖性能的养殖和野生maraena白鱼(Coregonus maraena)和受精的精子激活三个激活率的解决方案(如):台球解决方案(BS: 20 毫米三羟甲基氨基甲烷含液1 毫米CaCl₂,30 mM甘氨酸,和154年 mM氯化钠;pH值9.0;330年同渗重摩 mOsm·kg⁻¹),生理盐水(SS: 20 毫米三羟甲基氨基甲烷包含30 mM氯化钠;液pH值8.5;80年 mOsm·kg⁻¹),和湖水(LW: pH值7.5;20 mOsm·kg⁻¹)。尽管养殖鱼和野生鱼在基本生殖性状（卵重、假性腺-体细胞指数、精子体积、精子产量）方面没有显著差异，但在精子动力学和受精成功率方面存在显著差异。养殖和野生白鱼精子激活所用的AS不同，其进行性运动（PRG）、曲线速度（VCL）、直线速度（VSL）、摆动（WOB）和搏动交叉频率（BCF）均有显著变化。施用BS后，养殖鱼与野生鱼的受精率无显著差异。然而，与养殖鱼相比，在野生鱼中，SS和LW的受精率更高，尽管在这些条件下精子活力较低。线性回归分析显示，LW激活后的野生雄性高精子速度与受精成功率呈显著负相关，表明过度的运动可能会过早消耗精子能量储备。这些发现突出表明，最佳运动而不是最大运动可能对白鱼（特别是野生种群）的成功受精更为关键。然而，这一假设需要进一步的研究。

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引用次数: 0

Maternal placental mitochondrial dysfunction, oxidative stress, and autophagy by regulating ferroptosis in pregnant ewe model 母羊妊娠模型中母体胎盘线粒体功能障碍、氧化应激和自噬的调节

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2026-01-01 DOI: 10.1016/j.anireprosci.2025.108096

Feiyang He , Huisi Wu , Gao Liu , Jianing Wang , Bei Zhang , Xingyu Du , Mabrouk Elsabagh , Mengzhi Wang , Hao Zhang

Elevated prenatal testosterone (T) induces placental insufficiency and fetal growth restriction (FGR) in sheep, a process hypothesized to involve oxidative stress (OS), mitochondrial dysfunction, autophagy, and ferroptosis. While ferroptosis is recognized as a significant contributor to placental pathophysiology, its specific role in T-mediated ovine placental dysfunction required further investigation. To address this, an in vivo study was conducted wherein pregnant Hu sheep received intramuscular injections of 100 mg T propionate or a control vehicle twice weekly from gestational day (GD) 60–130. Complementarily, in vitro experiments utilized dihydrotestosterone (DHT)-exposed ovine trophoblast cells (OTCs), which were further treated with the ferroptosis activator Erastin or inhibitor Ferrostatin-1 (Fer-1) to directly probe the functional impact of ferroptosis. Our results demonstrated that T administration in vivo recapitulated the pathological phenotype, triggering placental OS, mitochondrial dysfunction, ferroptosis, autophagy, and culminating in FGR. Consistent with these findings, DHT exposure in OTCs induced a similar suite of cellular stresses, including OS, mitochondrial impairment, ferroptosis, and autophagy. Crucially, the inhibition of ferroptosis with Fer-1 in DHT-treated OTCs was found to attenuate these detrimental effects, notably alleviating OS, iron overload, mitochondrial dysfunction, and autophagic activity. Conversely, the co-administration of the ferroptosis inducer Erastin effectively abolished the protective changes conferred by Fer-1, thereby substantiating a central role for ferroptosis in the cascade of T-induced placental dysfunction.

产前睾酮(T)升高可导致绵羊胎盘功能不全和胎儿生长受限（FGR），这一过程可能涉及氧化应激（OS）、线粒体功能障碍、自噬和铁下垂。虽然铁下垂被认为是胎盘病理生理的重要因素，但其在t介导的绵羊胎盘功能障碍中的具体作用有待进一步研究。为了解决这个问题，进行了一项体内研究，从妊娠期（GD） 60-130开始，每周两次肌肉注射100 mg 丙酸T或对照物。此外，体外实验利用暴露于双氢睾酮（DHT）的羊滋养细胞（OTCs），进一步用铁下垂激活剂Erastin或抑制剂ferstat -1 （Fer-1）处理，直接探讨铁下垂对功能的影响。我们的研究结果表明，体内给药重现了病理表型，引发胎盘OS、线粒体功能障碍、铁下垂、自噬，并最终导致FGR。与这些发现一致的是，暴露在OTCs中的DHT诱导了一系列类似的细胞应激，包括OS、线粒体损伤、铁下垂和自噬。至关重要的是，在dht处理的OTCs中，发现用fe -1抑制铁下垂可以减轻这些有害影响，特别是减轻OS、铁过载、线粒体功能障碍和自噬活性。相反，铁下垂诱导剂Erastin的联合使用有效地消除了铁-1所赋予的保护性变化，从而证实了铁下垂在t诱导的胎盘功能障碍级联中的核心作用。

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引用次数: 0

High glucose in the extender impacts the metabolic phosphoproteome and modifies the phosphorylation of AKAP4 in stallion spermatozoa 扩展物中的高葡萄糖影响了种马精子的代谢磷酸化蛋白质组，并改变了AKAP4的磷酸化

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-12-30 DOI: 10.1016/j.anireprosci.2025.108095

Laura Becerro-Rey , Francisco Eduardo Martín-Cano , Antonio Silva-Rodríguez , Cristina Ortega-Ferrusola , Eva da Silva-Álvarez , Clara Zabalo-Palomo , Cruz Gil , Fernando J. Peña

Commercial extenders for equine semen typically contain high glucose concentrations; however, recent research indicates that these supraphysiological concentrations of glucose may be detrimental. Spermatozoa lack translational activity and depend on post-translational protein modifications for their regulation. Supraphysiological glucose concentrations are known to modify cell regulation, and we hypothesize that post-translational modification deregulation associated with supraphysiological glucose concentrations occurs in spermatozoa extended in high glucose media; and increasing the concentration of pyruvate in the media, may prevent this deregulation. Stallion ejaculates were split into different media containing 67 mM glucose and 1 mM pyruvate or 67 mM glucose and 10 mM pyruvate. After three hours of incubation at 38ºC, aliquots were taken for proteomic and kinematic analysis. Spermatozoa incubated in the high glucose (67 mM), low pyruvate (1 mM) media showed an alteration of the phosphorylation of the AKAP4 (P < 0.01), and modifications in gene ontology terms linked to sperm functionality. The incorporation of 10 mM pyruvate prevented all these changes, also kinematic efficiency was improved in the 67 mM glucose 10 mM pyruvate media. It is concluded that supraphysiological glucose concentrations alter the post-translational modifications of proteins essential for sperm function, and that this effect can be reduced in the presence of 10 mM pyruvate.

商用马精液填充剂通常含有高浓度葡萄糖；然而，最近的研究表明，这些超生理浓度的葡萄糖可能是有害的。精子缺乏翻译活性，依赖于翻译后蛋白修饰进行调节。已知超生理葡萄糖浓度会改变细胞调节，我们假设翻译后修饰的解除与超生理葡萄糖浓度相关，发生在高糖培养基中的精子中；而增加媒介中丙酮酸的浓度，可能会阻止这种放松管制。将种马射精分成含有67 mM葡萄糖和1 mM丙酮酸或67 mM葡萄糖和10 mM丙酮酸的不同培养基。38ºC孵育3小时后，取等分进行蛋白质组学和运动学分析。在高糖（67 mM）、低丙酮酸（1 mM）培养基中培养的精子显示出AKAP4磷酸化的改变（P <； 0.01），以及与精子功能相关的基因本体术语的修改。10 mM丙酮酸的掺入阻止了所有这些变化，并且在67 mM葡萄糖10 mM丙酮酸培养基中运动效率也有所提高。结论是，生理上的葡萄糖浓度改变了精子功能所必需的蛋白质的翻译后修饰，并且这种影响可以在10 mM丙酮酸存在下降低。

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引用次数: 0

Aquaporins in bovine sperm: Promising target for male reproductive strategy 牛精子中的水通道蛋白：雄性生殖策略的有希望的目标

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-12-30 DOI: 10.1016/j.anireprosci.2025.108091

Katarzyna Michałek , Sławomir Zych , Dariusz Gączarzewicz , Marta Marynowska , Patrycja Oberska-Łuszczewska , Andrzej Syczewski , Ernestats Mockus , Elena Bartkiene

The search for new indicators for assessing the quality of cryopreserved semen is necessary to reduce the number of ineffective insemination procedures in cattle. Aquaporins (AQPs) are small transmembrane proteins that facilitate the transport of water and other small solutes across the membrane. In this study we identified and localized AQP3, AQP7, AQP8 and AQP11 in frozen-thawed bull spermatozoa and investigated the relationship between their expression and specific semen quality parameters, as well as the concentrations of free amino acids (AA), biogenic amines (BA), and the composition of fatty acids (FA) in spermatozoa and seminal plasma. The collected cryopreserved semen samples were classified into two groups: high (H) and low (L) sperm motility. The analysis showed that the percentage of spermatozoa with intense AQP3 staining in the midpiece, and diffuse AQP7 staining in the post-acrosomal region and midpiece, was significantly higher in the H group. Furthermore, AQP3, AQP7 and AQP8 expression showed positive correlations with multiple semen quality parameters and concentrations of selected AA and FA. The results indicate that the analysis of aquaporin distribution in spermatozoa, particularly AQP3 and AQP7, may serve as a useful parameter for assessing the quality of cryopreserved semen in future studies. Detailed analysis of their location and expression allowed for the determination of the potential role of these proteins in the osmoadaptation process to the hypertonic environment of the uterus and fallopian tubes.

寻找新的指标来评估冷冻精液的质量是必要的，以减少牛的无效授精程序的数量。水通道蛋白（AQPs）是一种小的跨膜蛋白，促进水和其他小溶质在膜上的运输。本研究在冻融公牛精子中对AQP3、AQP7、AQP8和AQP11进行了鉴定和定位，并研究了它们的表达与精子和精浆中特定精液质量参数以及游离氨基酸（AA）、生物胺（BA）浓度和脂肪酸（FA）组成的关系。将收集的冷冻精液样本分为高(H)和低(L)精子活力两组。分析显示，H组精子中部AQP3染色强烈，顶体后区和中部AQP7染色弥漫性的比例显著高于H组。AQP3、AQP7和AQP8的表达量与精液质量参数及所选AA和FA浓度呈正相关。结果表明，分析精子中水通道蛋白的分布，特别是AQP3和AQP7的分布，可以作为未来研究中评估冷冻精液质量的一个有用参数。通过对其位置和表达的详细分析，可以确定这些蛋白在子宫和输卵管高渗环境的渗透适应过程中的潜在作用。

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引用次数: 0

Retrospective evaluation of the reproductive status of male grey wolves (Canis lupus) in Germany based on animals found dead: Influence of age, body weight and season 基于死亡动物对德国雄性灰狼（Canis lupus）生殖状况的回顾性评价：年龄、体重和季节的影响

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-12-27 DOI: 10.1016/j.anireprosci.2025.108094

Karin Müller , Ilka Reinhardt , Alexandra Weber , Gesa Kluth , Guido Fritsch , Claudia A. Szentiks

Since the return of the grey wolf (Canis lupus) to Germany in the late 1990s, the Federal Documentation and Consultation Centre on Wolves has been conducting nationwide population monitoring, with animals found dead representing an important source of information. In addition to the standardized necropsy at the Leibniz Institute for Zoo and Wildlife Research, 182 male wolves were examined for their reproductive status between 2020 and 2024. Testis mass was measured and spermatogenic activity was determined by flow cytometric ploidy analyses of dissociated testis tissue and the presence of epididymal sperm. These reproductive parameters were evaluated with regard to age class (juvenile, subadult, adult) and month of death (seasonality). Deciphering seasonality in subadult and adult males showed that haploid cells and epididymal sperm are not only present during the mating season (January to March) but also before (October to December), as meiotic activity already begins in late summer. Almost no spermatogenic activity was detected between May and August. While the body mass of (sub)adult males remained relatively constant throughout the year, testis/body mass ratio increased with spermatogenic activity. Spermatogenic activity already occurred in juvenile males, but it was delayed, and the average level of reproductive parameters increased from juvenile to subadult males, reaching their maximum in adult wolves. Sperm production varied greatly among juveniles and was related to their body and testis mass. Basic sexual maturity was only reached at a body mass of more than 25 kg and a testis mass of more than 5 g.

自从灰狼（Canis lupus）在20世纪90年代末回归德国以来，联邦狼文件和咨询中心一直在进行全国范围内的种群监测，发现死亡的动物是一个重要的信息来源。除了莱布尼茨动物园和野生动物研究所的标准化尸检外，在2020年至2024年期间，还对182只雄性狼的生殖状况进行了检查。通过对游离睾丸组织和附睾精子进行流式细胞术倍体分析，测定睾丸质量和生精活性。这些生殖参数根据年龄类别（幼鱼、亚成虫、成虫）和死亡月份（季节性）进行评估。对亚成年和成年雄鱼的季节性分析表明，单倍体细胞和附睾精子不仅在交配季节（1月至3月）存在，而且在交配季节（10月至12月）之前也存在，因为减数分裂活动在夏末就已经开始了。5月至8月间几乎没有发现生精活动。虽然（亚）成年雄性的体重全年保持相对恒定，但睾丸/体重比随着生精活动的增加而增加。幼雄的生精活动已经发生，但发生时间较晚，生殖参数的平均水平从幼雄到亚成年雄增加，在成年狼中达到最大值。幼鱼的精子产量差异很大，并与它们的身体和睾丸质量有关。只有当体重超过25 kg，睾丸质量超过5 g时，才能达到基本的性成熟。

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引用次数: 0

Temporal trends in porcine and bovine semen characteristics: A retrospective long-term analysis of semen production data 猪和牛精液特征的时间趋势：精液生产数据的回顾性长期分析

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-12-25 DOI: 10.1016/j.anireprosci.2025.108093

Britta Hensel , Sophie Henneberg , Filip Schröter , Maren Hürland , Jan-Hendrik Osmers , Stefan Geissendörfer , Matthias Lautner , Markus Jung , Martin Schulze

Artificial insemination (AI) is frequently used in cattle and pig production, and semen quality is crucial to both genetic advancement and successful reproduction. While several studies have documented deteriorating semen quality in humans, dogs, and horses, knowledge about long-term trends of bull and boar semen quality is insufficient despite its biological and economic significance. The present study aimed to assess temporal trends in semen quality parameters in bulls and boars over a period of 18 and 22 years, respectively, using production data collected at a single German AI center for each species. A retrospective analysis of semen characteristics, including ejaculate volume, sperm concentration, total sperm motility, and sperm output, was performed on 47,757 bull ejaculates collected between 1997 and 2019 and 619,368 boar ejaculates collected between 2005 and 2023. Temporal trends were evaluated using linear regressions. The analysis revealed a significant increase in sperm concentration and total sperm motility for both bulls and boars. The decrease in ejaculate volume in both species did not negatively affect overall sperm output, which showed an increasing trend in line with sperm concentration and total sperm motility for bulls as well as boars. All observed trends were highly significant (P < 0.001). Results contrast findings in other species and highlight the efficiency of breeding strategies in the cattle and pig industry, improving not only production traits but also reproductive performance.

人工授精（AI）经常用于牛和猪的生产，精液质量对遗传进步和成功繁殖至关重要。虽然有几项研究证明人类、狗和马的精液质量正在恶化，但关于公牛和野猪精液质量的长期趋势的知识还不够，尽管它具有生物学和经济意义。本研究旨在利用在德国单个人工智能中心收集的每个物种的生产数据，分别评估公牛和公猪在18年和22年期间精液质量参数的时间趋势。对1997年至2019年收集的47,757份公牛射精和2005年至2023年收集的619,368份野猪射精进行了精液特征的回顾性分析，包括射精量、精子浓度、精子总活力和精子输出量。使用线性回归评估时间趋势。分析显示，公牛和公猪的精子浓度和总精子活力显著增加。射精量的减少并没有对公牛和公猪的精子总量产生负面影响，精子总量的增加趋势与精子浓度和总精子活动力一致。所有观察到的趋势都非常显著（P <； 0.001）。结果对比了其他物种的发现，突出了在牛和猪行业中育种策略的效率，不仅提高了生产性状，而且提高了繁殖性能。

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MicroRNA-150 deficiency promotes progesterone synthesis and apoptosis in goat luteal cells by targeting nuclear receptor NR4A1 MicroRNA-150缺乏通过靶向核受体NR4A1促进山羊黄体细胞的黄体酮合成和凋亡

IF 3.3 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science

Pub Date : 2025-12-25 DOI: 10.1016/j.anireprosci.2025.108092

Hao Yu, Xiaotong Li, Xianyi Zhou, Jie Zhao, Wei Wang, Dagan Mao

This study aimed to investigate the effects of microRNA-150 (miR-150) on progesterone (P4) synthesis and apoptosis in goat luteal cells. Corpus luteum (CL) tissues were collected from non-pregnant goats. qPCR and in situ hybridization showed that miR-150 was highly expressed in the early CL but decreased in mid and late CL, and miR-150 was positively localized in luteal cells. Luteal cells from mid-cycle CL were then transfected with Agomir-150 (mimics) or Antagomir-150 (inhibitor). Immunoassays and qPCR results showed that Agomir-150 suppressed P4 levels and down-regulated the mRNA and protein expressions of hormone‑sensitive lipase (HSL), phosphorylated HSL (p-HSL), steroidogenic acute regulatory protein (StAR), and 3β‑hydroxysteroid dehydrogenase (HSD3B), whereas Antagomir-150 had the opposite effects. In addition, flow cytometry, qPCR, and Western blot results showed that Agomir-150 had no significant impact on cell apoptosis, while Antagomir-150 promoted apoptosis, up-regulated the mRNA and protein expressions of BCL2-associated X protein (BAX), Fas cell surface death receptor (FAS), tumor necrosis factor receptor 1 (TNFR1), Caspase-3, cleaved Caspase-3, and down-regulated B-cell lymphoma 2 (BCL-2) expression. Bioinformatic analysis identified nuclear receptor subfamily 4, group A, member 1 (NR4A1) as a potential miR-150 target. Dual-luciferase assays confirmed that miR-150 directly binds to the NR4A1–3′UTR and represses its expression, which was further validated by qPCR and Western blot. A rescue experiment revealed that NR4A1 knockdown partially reversed the Antagomir-150–induced up-regulation of P4 synthesis and apoptosis. Overall, this study demonstrates that miR-150 contributes to P4 synthesis and apoptosis in goat luteal cells by targeting NR4A1.

本研究旨在探讨microRNA-150 （miR-150）对山羊黄体细胞中孕酮（P4）合成和凋亡的影响。采集未怀孕山羊的黄体组织。qPCR和原位杂交显示miR-150在CL早期高表达，在CL中晚期表达降低，且miR-150在黄体细胞中呈阳性定位。然后用Agomir-150（模拟物）或Antagomir-150（抑制剂）转染中期CL的黄体细胞。免疫分析和qPCR结果显示，Agomir-150抑制P4水平，下调激素敏感脂肪酶（HSL）、磷酸化HSL （p-HSL）、类固醇急性调节蛋白（StAR）和3β羟基类固醇脱氢酶（HSD3B） mRNA和蛋白表达，而Antagomir-150则相反。此外，流式细胞术、qPCR和Western blot结果显示，Agomir-150对细胞凋亡无显著影响，而Antagomir-150促进细胞凋亡，上调bcl2相关X蛋白（BAX）、Fas细胞表面死亡受体（Fas）、肿瘤坏死因子受体1 （TNFR1）、Caspase-3、cleaved Caspase-3 mRNA和蛋白表达，下调b细胞淋巴瘤2 （BCL-2）表达。生物信息学分析发现核受体亚家族4，A组，成员1 （NR4A1）是miR-150的潜在靶点。双荧光素酶实验证实miR-150直接结合NR4A1-3'UTR并抑制其表达，qPCR和Western blot进一步验证了这一点。一项救援实验显示，NR4A1敲低部分逆转了antagomir -150诱导的P4合成上调和细胞凋亡。总之，本研究表明miR-150通过靶向NR4A1参与山羊黄体细胞P4的合成和凋亡。

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