Theriogenology最新文献

{"title":"Expression pattern of the HIFα-family in equine chorioallantois during pregnancy and placental pathology","authors":"Yatta Boakari ,&nbsp;Hossam El-Sheikh Ali ,&nbsp;Gustavo Henrique Doná Rodrigues Almeida ,&nbsp;Kirsten E. Scoggin ,&nbsp;Claudia Barbosa Fernandes ,&nbsp;Sophia Panelli Marchio ,&nbsp;Mats H.T. Troedsson ,&nbsp;Ana Claudia Oliveira Carreira","doi":"10.1016/j.theriogenology.2025.117714","DOIUrl":"10.1016/j.theriogenology.2025.117714","url":null,"abstract":"<div><div>Equine placental dysfunction impairs fetal growth and disrupts pregnancy outcomes. Despite many advances in diagnosis, the molecular pathophysiology of equine placentitis and premature placental separation remains poorly understood. However, the hypoxia-inducible factor (HIF-α) pathway is related to placental hypoxia, and its modulation in placental diseases has been shown in other species. Therefore, we hypothesized that in conditions marked by equine placental dysfunction, oxygen deprivation in the chorioallantois membrane triggers the activation of the HIF-α pathway, which is related to molecular alterations associated with placental insufficiency. Accordingly, we compared the expression of <em>HIF1A, HIF2A, HIF3A,</em> and <em>HIF1AN</em> genes in equine chorioallantois 1) at various points during normal gestation, and in mares with 2) ascending placentitis (AP), 3) nocardioform placentitis (NP), and 4) premature placental separation (PPS), compared to mares with normal pregnancies (controls). During normal pregnancy, expression of HIF-related genes remained low until a significant post-partum increase, suggesting a balanced regulatory state under normoxic conditions. In contrast, the three pathological conditions exhibited a distinct expression profile: AP was marked by <em>HIF1A</em> upregulation and concurrent suppression of <em>HIF3A</em> and <em>HIF1AN</em>; NP showed reduced <em>HIF2A</em> and <em>HIF3A</em> expression without changes in <em>HIF1A</em>; and PPS was characterized by <em>HIF1A</em> upregulation with <em>HIF1AN</em>, <em>HIF2A,</em> and <em>HIF3A</em> downregulation. Our findings reveal that although AP, NP, and PPS all involve oxygen deprivation, the molecular hypoxia response is disease specific. This study provides novel insight into the equine placental hypoxic response. This warrants further research to elucidate these mechanisms and explore them as potential diagnostic targets for equine placental dysfunctions.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117714"},"PeriodicalIF":2.5,"publicationDate":"2025-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145356305","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel long noncoding RNA lnc_2217 regulates zygotic genome activation through miR-877-3p/YTHDF2 pathway in goat","authors":"Haonan Chen,&nbsp;Mengyuan Tian,&nbsp;Ershuai Kuang,&nbsp;Xiaowei Chen,&nbsp;Yingnan Yang,&nbsp;Mingtian Deng","doi":"10.1016/j.theriogenology.2025.117713","DOIUrl":"10.1016/j.theriogenology.2025.117713","url":null,"abstract":"<div><div>Precisely timed zygotic genome activation (ZGA) and controlled maternal mRNA decay are critical for early embryo development in mammals. Although long non-coding RNA (lncRNAs) have been reported to regulate the early embryo development, their roles remain unclear during ZGA. Here we show that novel lncRNA lnc_2217 is indispensable for the early embryo development as microinjection of siRNAs against lnc_2217 reduced blastocyst formation from 32.5 % to 13.9 % and increased developmental arrest at the 8- and 16-cell stages. RNA-seq and 5-EU incorporation assays revealed that lnc_2217 knockdown impaired global de-novo transcription and provoked over-degradation of maternal mRNAs at the 8-cell stage. Mechanistically, lnc_2217 functions as a competing endogenous RNA that sequesters miR-877-3p, thereby relieving repression of the m6A reader YTHDF2. Loss of lnc_2217 leads to YTHDF2 down-regulation and accelerated turnover of key maternal transcripts (ZAR1, YAP1, UHRF1). Our findings identify an lnc_2217/miR-877-3p/YTHDF2 axis that synchronizes transcriptional activation with selective maternal mRNA clearance during ZGA, highlighting lnc_2217 as a crucial regulator of early goat embryogenesis.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117713"},"PeriodicalIF":2.5,"publicationDate":"2025-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145321706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanoplastics internalization impairs mitochondrial activity in equine sperm","authors":"Sofia Dindo ,&nbsp;Laura Tovar-Pascual ,&nbsp;Vito Antonio Baldassarro ,&nbsp;Diego Bucci ,&nbsp;Beatrice Mislei ,&nbsp;Marcella Spinaci ,&nbsp;Jose Manuel Ortiz-Rodriguez","doi":"10.1016/j.theriogenology.2025.117711","DOIUrl":"10.1016/j.theriogenology.2025.117711","url":null,"abstract":"<div><div>Nanoplastics (NP) accumulation in biological tissues and their adverse effects on fertility through inflammatory and oxidative stress responses have recently been described as consequences of global plastic pollution. However, little is known about the impact of NP on gametes. This study aimed to assess the internalization of NP and their effects on mature equine spermatozoa. Frozen-thawed ejaculates from five stallions were divided into untreated control (CTR) and samples supplemented with different concentrations (10, 50, 100 and 200 μg/mL) of 30 nm polystyrene NP. At baseline (T0), and after 1 (T1) and 3 h (T3) of incubation at 38 °C, sperm viability, mitochondrial activity, and intracellular ROS were evaluated by flow cytometry, while sperm motility was assessed using a CASA system. NP internalization was analyzed by confocal microscopy and flow cytometry using fluorescent NP. Results showed that NP were internalized by live spermatozoa, accumulating in the post-acrosomal and/or the mid piece region. NP exposure led to reduced sperm viability (CTR T1: 50.7 ± 14.0 % vs 200 μg/mL T1: 36.6 ± 11.8 %, <em>p&lt;0.05</em>), decreased mitochondrial activity (CTR T1: 35.4 ± 15.8 % vs 200 μg/mL T1: 16.3 ± 14.0 %, <em>p&lt;0.001</em>), and increase proportions of live sperm with high intracellular O₂⁻· levels (CTR T1: 39.0 ± 10.1 % vs 200 μg/mL T1: 47.3 ± 11.0 %, <em>p&lt;0.05</em>). These results allow us to conclude that equine sperm quality may be compromised by nanoplastics internalization, which pre-eminently impairs mitochondrial activity. This research furnishes some bases for further studies on the potential implications of NP exposure for fertility.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117711"},"PeriodicalIF":2.5,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145309309","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Association of colostrum traits with the farrowing process and reproductive performance of sows in a free farrowing system","authors":"Lea Käser ,&nbsp;Laura Jahn ,&nbsp;Rupert Bruckmaier ,&nbsp;Alexander Grahofer","doi":"10.1016/j.theriogenology.2025.117708","DOIUrl":"10.1016/j.theriogenology.2025.117708","url":null,"abstract":"<div><div>Colostrum is vital for neonatal piglets, supplying energy, immunoglobulins, hormones, and other bioactive compounds critical for early survival. Beyond its nutritional role, colostrum also reflects the sow's physiological and hormonal state during the periparturient period. This study investigated associations between colostrum composition, farrowing traits, and reproductive performance in a free farrowing system. Sixty sows were monitored, and colostrum samples (5–10 mL) were collected after the birth of the first piglet. Concentrations of progesterone, albumin, and alpha-lactalbumin were measured in colostrum serum. Farrowing characteristics and litter outcomes were recorded.</div><div>Across all sows, 897 piglets were born and the mean farrowing duration was 242 min. Colostrum progesterone averaged 21.6 ± 10.8 ng/mL, albumin 7.33 ± 1.47 mg/mL, and alpha-lactalbumin 1602.7 ± 506.9 ng/mL. Progesterone concentrations were positively correlated with albumin (r = 0.29; p = 0.02) and alpha-lactalbumin (r = 0.38; p = 0.002). Progesterone also correlated positively with log-transformed farrowing duration (r = 0.27; p = 0.03), piglet to placenta expulsion duration (r = 0.26; p = 0.04), and incidence of severe meconium staining (r = 0.27; p = 0.04). Sows requiring more than one obstetrical intervention had significantly higher progesterone concentrations (25.9 ng/mL) compared to those with none or one intervention (19.3 ng/mL; p = 0.02).</div><div>This study is the first to link specific colostrum traits with farrowing outcomes in a free farrowing system, suggesting colostrum progesterone, despite its low correlation, may serve as an indicator for prolonged farrowing.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117708"},"PeriodicalIF":2.5,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145303652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Influence of hydroxytyrosol in semen extenders on fertilizing capacity and embryo development using cooled and cryopreserved bull sperm","authors":"Marianela Balbi ,&nbsp;Ana Laura Flaherti ,&nbsp;Trinidad Otero Fernández ,&nbsp;Nicolás Agustín Farnetano ,&nbsp;Juan Mateo Anchordoquy ,&nbsp;Juan Patricio Anchordoquy","doi":"10.1016/j.theriogenology.2025.117702","DOIUrl":"10.1016/j.theriogenology.2025.117702","url":null,"abstract":"<div><div>This study investigated the effects of hydroxytyrosol (HT), a potent antioxidant polyphenol, on sperm functionality, fertilizing ability, and embryo development when added to semen extenders using cooled and cryopreserved bull semen. In a preliminary trial, 1 and 25 μM HT were identified as the most effective concentrations and subsequently used in the main experiments. Semen from 12 Angus bulls was divided into aliquots and stored either refrigerated at 4 °C for 24 h or cryopreserved. Sperm parameters (motility, viability, membrane and acrosomal integrity), malondialdehyde (MDA) levels, total antioxidant capacity (TAC), zona pellucida (ZP) binding, and <em>in vitro</em> fertilization outcomes were assessed. In cooled semen, HT enhanced functional membrane integrity and acrosomal status, and increased sperm–ZP binding. Supplementation with 1 μM HT improved embryo quality, increasing hatching rate and blastocyst cell number. In cryopreserved semen, 25 μM HT improved ZP-binding capacity, blastocyst rates, and blastocyst cell numbers. These findings suggest that HT supplementation improves sperm functionality and embryo development under both cooled and cryopreserved storage conditions, highlighting its potential to enhance assisted reproductive technologies in cattle.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117702"},"PeriodicalIF":2.5,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145281215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lipid droplets-mitochondria interaction involved in testosterone synthesis in Leydig cells of dairy goats (Capra hircus)","authors":"Fang-e Zhao ,&nbsp;Shuo Wang ,&nbsp;Xinge Zhang ,&nbsp;Haorui Yang ,&nbsp;Mengyuan Zhou ,&nbsp;Hongbo Chen ,&nbsp;Na Chen ,&nbsp;Yongjie Wu ,&nbsp;Shulin Chen ,&nbsp;Jun Liu ,&nbsp;Hong Chen","doi":"10.1016/j.theriogenology.2025.117704","DOIUrl":"10.1016/j.theriogenology.2025.117704","url":null,"abstract":"<div><div>Leydig cells (LCs) are the primary testosterone-producing cells. Their steroidogenic capacity is heavily influenced by the postnatal development of the LC lineage. The final developmental stage of LCs involves transition from immature LCs (ILCs) to adult LCs (ALCs) during puberty. However, the morphological and functional changes that occur during this transition in most mammals remain unclear. In this study, we conducted a comparative analysis of morphological and functional changes in developing LCs in dairy goats. The results revealed that during the transition from ILC to ALC, the ovoid LC transformed into an irregular round shape with a well-developed smooth endoplasmic reticulum (sER), numerous mitochondria, and small lipid droplets (LDs) distributed in the cytoplasm. Subsequently, the isolated primary ILCs were matured using chorionic gonadotropin (CG). CG stimulation increases the expression of steroidogenic genes and decreases the levels of testosterone-metabolizing enzymes. In vitro studies have shown that small LDs are the preferred source of cholesterol substrates in steroidogenesis, and that their interaction with mitochondria facilitates free cholesterol transport. Further analyses using confocal microscopy, ultrastructural analysis, and co-immunoprecipitation consistently demonstrated that vimentin filaments contributed to the interaction between LDs and mitochondria. Specifically, the knockdown of vimentin using siRNA substantially reduced the physical contact between LDs and mitochondria, resulting in inadequate cholesterol transport and ultimately impaired steroidogenesis. Overall, our findings indicate that significant morphological and functional changes occurred during the transition from ILCs to ALCs in goats. The cytoskeleton, primarily composed of vimentin filaments, is an important component of the interactions between LDs and mitochondria and contributes to cholesterol transport during testosterone synthesis.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117704"},"PeriodicalIF":2.5,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145293891","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lactate metabolism inhibition disrupts goat oocyte meiosis via H3K18la-mediated epigenetic dysregulation","authors":"Dongxu Li ,&nbsp;Zhixuan Zhou ,&nbsp;Minghui Zhu ,&nbsp;Honglei Fan ,&nbsp;Xinsong Xu ,&nbsp;Honghui Lu ,&nbsp;Feng Wang ,&nbsp;Dagan Mao ,&nbsp;Ziyu Wang ,&nbsp;Yongjie Wan","doi":"10.1016/j.theriogenology.2025.117706","DOIUrl":"10.1016/j.theriogenology.2025.117706","url":null,"abstract":"<div><div>Lactylation, a newly discovered histone post-translational modification, has emerged as a key metabolic-epigenetic regulator; however, its dynamic functions during oocyte maturation remain unexplored. This study demonstrates that pharmacological inhibition of lactate metabolism by GNE-140 (a lactate dehydrogenase inhibitor) and 2-deoxy-D-glucose (2-DG, a glycolysis inhibitor) significantly compromises the quality of goat oocytes. These inhibitors disrupt meiotic progression and impair developmental competence by perturbing spindle assembly, actin cytoskeleton dynamics, and chromosomal alignment, ultimately leading to maturation arrest. In addition, metabolic inhibition induces pronounced mitochondrial dysfunction, characterized by elevated levels of reactive oxygen species (ROS), increased DNA damage, and apoptosis. Functionally, oocytes exposed to these metabolic inhibitors exhibit markedly reduced fertilization rates and diminished blastocyst formation potential, highlighting the long-term developmental consequences of impaired lactate metabolism. To elucidate the underlying mechanisms, we integrated transcriptomic analysis (RNA-seq) with chromatin profiling (CUT&amp;Tag for H3K18la), which revealed ribosomal protein L8 (RPL8) as a potential downstream effector. Pathway enrichment analysis further implicated disruptions in oxidative phosphorylation, lactate metabolism, and gene networks associated with oocyte meiosis. Collectively, our findings uncover a novel mechanistic link between metabolic regulation, epigenetic modification, and oocyte maturation, providing valuable insights for optimizing <em>in vitro</em> maturation protocols in caprine species and potentially other mammals.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117706"},"PeriodicalIF":2.5,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145293856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Estradiol treatment and double dose of GnRH for resynchronization protocols and timed artificial insemination in lactating dairy cows","authors":"Juan Carlos Tschopp ,&nbsp;Alejo Menchaca ,&nbsp;Gabriel A. Bó","doi":"10.1016/j.theriogenology.2025.117707","DOIUrl":"10.1016/j.theriogenology.2025.117707","url":null,"abstract":"<div><div>The aim of this study was to compare pregnancy per AI (P/AI) in resynchronization (Resynch) protocols using different hormonal combinations (estradiol benzoate [EB], a double dose of GnRH, or EB with a double dose of GnRH) administered on the day of progesterone (P4) device insertion. We hypothesized that combined administration of EB and GnRH on Day 25 after the first timed AI (TAI) is more efficacious than the administration of EB or GnRH alone in the resynchronization of estrus and ovulation in lactating dairy cows. The secondary objective was to compare P/AI in cows treated with two different GnRH agonists (gonadorelin acetate or buserelin acetate) on the day of P4 device insertion. This study included lactating Holstein cows (382 in their first lactation and 629 in their second or more lactations), averaging 135.4 ± 15.6 days in milk, with a body condition score of 2.8 ± 0.1, and producing 41.3 ± 0.5 kg of milk per day. On Day 25 post first timed AI (TAI), all cows received a previously used CIDR-B device (1.9 g P4) and were randomly assigned to receive one of the following treatments: 2-mg EB, a double dose of GnRH, or a combination of 2-mg EB and a double dose of GnRH. Cows receiving GnRH were randomly divided into the 200-μg gonadorelin acetate group or 20-μg buserelin acetate group. On Day 32, all cows were examined via ultrasonography, and those that were not pregnant received 500 μg of cloprostenol (PGF2α). On Day 33, the CIDR-B devices were removed, and all cows received a second dose of PGF2α, 1 mg of estradiol cypionate (ECP), and 400 IU eCG, with all cows receiving tail paint for estrus detection. Cows that demonstrated estrus at 48 h were inseminated at that time, whereas those that did not received 100 μg of gonadorelin and were inseminated at 56 h. Cows in the EB + double-dose GnRH group exhibited greater estrus expression (93.6 %, <em>P</em> &lt; 0.05) than those in the EB (88.1 %) or double-dose GnRH (92.0 %) group, but no difference in the P/AI was observed among the groups (EB = 44.1 %, double-dose GnRH = 49.0 %, EB + double-dose GnRH = 49.0 %; <em>P</em> = 0.94). Similarly, no significant difference in the P/AI was observed between cows treated with gonadorelin acetate (50.0 %) and those treated with buserelin acetate (48.1 %, <em>P</em> = 0.31). In conclusion, the combined administration of EB and a double dose of GnRH does not improve P/AI compared with the administration of either hormone alone. Furthermore, both agonists produced comparable P/AI, confirming that they are equally effective when administered during P4 device insertion.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117707"},"PeriodicalIF":2.5,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145309265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Breed-specific testicular ultrasound profiles and pubertal markers in Bos indicus and composite bulls","authors":"Joedson Dantas Gonçalves ,&nbsp;Maria Emilia Franco Oliveira ,&nbsp;Rubens Paes de Arruda ,&nbsp;Cintia Righetti Marcondes ,&nbsp;Patrícia Tholon ,&nbsp;Felipe Zandonadi Brandão ,&nbsp;Gabriela Novais Azevedo ,&nbsp;Giovanna Galhardo Ramos ,&nbsp;Gabriel Brun Vergani ,&nbsp;Alexandre Rossetto Garcia","doi":"10.1016/j.theriogenology.2025.117705","DOIUrl":"10.1016/j.theriogenology.2025.117705","url":null,"abstract":"<div><div>Ultrasonography has offered potential to detect breed-specific differences and identify early puberty biomarkers in bulls. This study aimed to (a) characterize testicular ultrasound parameters in young bulls of the Nelore (<em>Bos indicus</em>) and Canchim (composite: 5/8 <em>Bos taurus</em> × 3/8 <em>Bos indicus</em>) breeds, and (b) identify potential ultrasonographic biomarkers associated with the pre- and post-pubertal phases. A total of 46 young (Nelore, n = 22; Canchim, n = 24) were evaluated using testicular biometry, B-mode ultrasonography, color Doppler of the parenchyma and pampiniform plexus, and spectral Doppler of the supratesticular artery. Semen analysis and serum testosterone measurements were conducted to determine pubertal status. Monthly collections were performed with longitudinal monitoring from 8 to 19 months of age. Subsequently, the animals were grouped into pre-pubertal and post-pubertal cohorts. Statistical analyses were performed using PROC MIXED in SAS, with significance set at P &lt; 0.05. Significant temporal increases were observed in scrotal circumference (P &lt; 0.001), testicular volume (P = 0.03), parenchymal echogenicity (P = 0.0007), and relative Doppler area (P = 0.0003). At most timepoints, Canchim bulls demonstrated consistently higher values than Nelore bulls. Notably, pre-pubertal Canchim bulls exhibited greater testicular echogenicity compared to their post-pubertal counterparts (P = 0.03), while pre-pubertal Nelore bulls showed reduced Doppler perfusion in the testicular parenchyma relative to post-pubertal individuals (P = 0.03). This study highlights significant breed-related differences in testicular ultrasound parameters during sexual maturation. Canchim bulls exhibited higher echogenicity and testicular perfusion compared to Nelore bulls. Furthermore, ultrasonographic features such as echogenicity and Doppler area emerged as promising, non-invasive biomarkers for monitoring pubertal development in beef cattle.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117705"},"PeriodicalIF":2.5,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145313854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ProAKAP4 as a protein marker of sperm quality: Insights from frozen-thawed ejaculated and epididymal sperm","authors":"Myrian Megumy Tsunokawa Hidalgo ,&nbsp;Ana Beatriz Marques de Almeida ,&nbsp;Leticia Amanda dos Santos Silva ,&nbsp;Vinicius Wagner Silva ,&nbsp;Julia Rodrigues Greghi ,&nbsp;Natália Ribeiro Sambatti ,&nbsp;Luiz Guilherme Corsi Trautwein ,&nbsp;Rodrigo Yudi Palhaci Marubayashi ,&nbsp;João Carlos Pinheiro Ferreira ,&nbsp;Maria Isabel Mello Martins","doi":"10.1016/j.theriogenology.2025.117698","DOIUrl":"10.1016/j.theriogenology.2025.117698","url":null,"abstract":"<div><div>The AKAP4 protein is the most abundant protein of the sperm fibrous sheath. This protein is considered instrumental in regulating sperm flagellum motion and its precursor, proAKAP4, is considered a biomarker for sperm quality in different species. Therefore, this study aimed to evaluate the relationship of proAKAP4 concentration on feline sperm kinematic, morphology, and membrane integrity of cryopreserved ejaculated and epididymal sperm. A total of fifteen adult mixed breed male cats between 1 and 3 years of age, with a mean weight of 4.21 ± 0.55 kg were included in this study. The first step of the study involved semen collection by urethral catheterization, using a sterile probe. In the second step, sperm were recovered from the epididymal tail after elective orchiectomy. The samples were cryopreserved in a TRIS-yolk extender. Thawed sperm straws were analyzed regarding their sperm kinematic, morphology and plasma membrane integrity. Sperm proAKAP4 concentration was determined using a commercial proAKAP4 ELISA kit. The relationship between proAKAP4 concentrations and sperm kinematics were evaluated by linear regressions, considering the proAKAP4 concentrations as independent variable and kinematic parameters as dependent variable. To compare concentration of proAKAP4 protein in both collection methods, paired <em>t</em>-test was used (P &lt; 0.05). When comparing proAKAP4 concentrations in ejaculated and epididymal sperm, it was observed that the mean of proAKAP4 in ejaculated sperm were higher, 31.43 ± 3.36 ng/mL (15.26–59.18 ng/mL) than epididymal sperm 19.58 ± 1.67 ng/mL (12.15–32.14 ng/mL) (P = 0.004). It was possible to observe the relationship between proAKAP4 concentration and the parameters: TM (r2 = 0.574, P = 0.001) and dance (r2 = 0.267, P = 0.049) in ejaculated sperm. Also, a relationship between the proAKAP4 concentration and TM (r2 = 0.670, P &lt; 0.001) and the percentage of static sperm cells (r2 = 0.477, P = 0.004) in epididymal sperm was seen. In conclusion, our findings indicated that proAKAP4 concentrations are correlated with kinematic parameters in both ejaculated and epididymal sperm post-thaw, suggesting its role in sustaining flagellar architecture and modulating capacitation-associated motility of felines.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"250 ","pages":"Article 117698"},"PeriodicalIF":2.5,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145270740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}