Theriogenology最新文献

{"title":"Metabolome and oxidative stress markers in the seminal plasma of Holstein bulls and their relationship with the characteristics of fresh and frozen/thawed sperm","authors":"E. Giaretta ,&nbsp;A. Damato ,&nbsp;L. Zennaro ,&nbsp;V. Bonfatti ,&nbsp;B. Mislei ,&nbsp;V. Vigolo ,&nbsp;M.E. Falomo ,&nbsp;F. Bertuzzo ,&nbsp;G. Gabai ,&nbsp;D. Bucci","doi":"10.1016/j.theriogenology.2025.01.015","DOIUrl":"10.1016/j.theriogenology.2025.01.015","url":null,"abstract":"<div><div>Seminal plasma composition has important role in sperm functionality and its freezability. The objective of this study was to test the hypothesis that seminal plasma (SP) oxidative status and metabolome are associated with fresh semen characteristics and freezability of bull sperm. To accomplish this objective, oxidative status markers and metabolome of SP of ejaculates obtained from 20 Holstein bulls (3 for each bull) were analyzed using spectrophotometry and nuclear magnetic resonance (¹H NMR). The ejaculates were classified into higher motility fresh semen (HMF) and lower motility fresh semen (LMF), according to total motility (TM) and progressive motility (PM) values of fresh semen. Then the ejaculates was cryopreserved and assigned to higher motility thawed group (HMT) or lower motility thawed group (LMT) according to TM and PM at 0 h post-thawing. Multivariate analyses were performed to identify the association between the functional characteristics of fresh and thawed semen and the SP parameters, in terms of the oxidative status and the metabolomic composition. According to our results, the advanced oxidative protein products (AOPP) and thiol concentrations in SP are significantly related to some physiological characteristics of the thawed sperm, such as higher viability, TM, PM and LIN and lower mitochondrial and cytoplasmic superoxide production in viable thawed cells. In contrast, a higher amount of C in the SP was negatively related to TM and PM of thawed semen and was associated with higher mitochondrial and cytoplasmic superoxide production.</div><div>In addition, partial least squares-discriminant analysis (PLS-DA) performed on the ¹H NMR spectra indicated a discrete separation between HMF and LMF groups, and good discrimination between HMT and LMT groups. Higher levels of formic acid, lactate, glycerol and phosphocholine, were found in the SP of the HMF group than in the LMF group. On the other hand, alanine, phenylalanine, and tyrosine were higher in the SP of the LMF group than in the HMF group. GABA, glutamate, histidine and glycerol were found in higher concentrations in the HMT group than in the LMT group, while fructose decreased in the HMT group. Our results showed that the oxidative and metabolomic status of SP is related to the physiological properties of semen and its freezability and open new fields in research of SP biomarkers of bull semen preservation and fertility.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 262-274"},"PeriodicalIF":2.4,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143075572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Porcine endometrial epithelial organoids: Generation, characterization, and the impact of stromal cells","authors":"Meng-Die Li,&nbsp;Hai-Yue Xu,&nbsp;Yu-Shi Chen,&nbsp;Xin-Cheng Zhang,&nbsp;Hua Ni","doi":"10.1016/j.theriogenology.2025.01.013","DOIUrl":"10.1016/j.theriogenology.2025.01.013","url":null,"abstract":"<div><div>Endometrial organoids (EOs) are three-dimensional models that emulate the endometrium, serving as an invaluable <em>in vitro</em> tool for investigating the cellular and molecular mechanisms underlying endometrial physiology and pathology during the estrous cycle and pregnancy. While significant progress has been made in the establishment and optimization of EOs for both humans and mice, research on such models in other species remains limited. This study aimed to develop porcine endometrial epithelial organoids (EEOs) to explore the regulatory mechanisms of uterine function and maternal-fetal interactions during porcine pregnancy, which are critical for enhancing reproductive efficiency and improving embryo transfer techniques. Utilizing a defined protocol involving Matrigel and a simplified culture medium, we successfully established EEOs from porcine endometrial epithelial cells, which demonstrated epithelial characteristics, proliferative capacity, glycogen secretion, and hormonal responsiveness. Co-culturing with porcine endometrial stromal cells not only enhanced EEO formation but also mitigated the negative effect of progesterone on glycogen secretion. This cost-effective and efficient method provides a valuable reference for EO generation in other large animal species.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 175-183"},"PeriodicalIF":2.4,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024774","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":"Supplementation with N-Acetyl-L-cysteine during in vitro maturation improves goat oocyte developmental competence by regulating oxidative stress","authors":"Yanfei Wang ,&nbsp;Qingwei Wang ,&nbsp;Quan Ji ,&nbsp;Pengcheng An ,&nbsp;Xiaodong Wang ,&nbsp;Yonghong Ju ,&nbsp;Ruiyang Li ,&nbsp;Yong Ruan ,&nbsp;Jiafu Zhao ,&nbsp;Maosheng Cao ,&nbsp;Xiang Chen","doi":"10.1016/j.theriogenology.2025.01.016","DOIUrl":"10.1016/j.theriogenology.2025.01.016","url":null,"abstract":"<div><div>Oocyte quality can affect mammal fertilization rate, early embryonic development, pregnancy maintenance, and fetal development. Oxidative stress induced by reactive oxygen species (ROS) is one of the most important causes of poor oocyte maturation <em>in vitro</em>. To reduce the degree of cellular damage caused by ROS, supplementation with the antioxidant N-Acetyl-L-cysteine (NAC) serves as an effective pathway to alleviate glutathione (GSH) depletion during oxidative stress. This study investigated the effects of NAC supplementation during <em>in vitro</em> maturation of goat oocytes and explored the molecular mechanisms of maturation by transcriptome sequencing of MⅡ oocytes. The results showed that 1.5 mM NAC significantly increased the rates of oocyte maturation and cumulus cell expansion and improved the subsequent development of embryos. During the subsequent culture of parthenogenetically activated embryos, 1.5 mM NAC significantly increased the division rate of oocytes and blastocyst rate. It also reduced the accumulation of ROS, increased the level of GSH, alleviated oxidative stress, and enhanced the antioxidant capacity and cell metabolic activity. Transcriptome sequencing results revealed that NAC treatment significantly increased the expression of <em>SIRT1</em>, <em>GGCT,</em> and <em>MITF</em> genes related to the cellular antioxidant system, as well as the <em>IDH3G</em> gene related to energy metabolism, and decreased the expression of <em>CASP8</em>, <em>FOS</em>, and <em>MMP1</em> genes related to apoptosis and cell invasion, as well as the <em>CCL2</em>. and <em>CXCL8</em> genes related to the inflammatory response. In conclusion, the findings suggest that NAC supplementation significantly reduces oxidative stress, improves antioxidant capacity and metabolic activity, promotes oocyte maturation, and improves oocyte quality.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 221-230"},"PeriodicalIF":2.4,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041630","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":"miRNA-125a regulates porcine oocyte maturation in vitro by targeting ADAR","authors":"Yue Han ,&nbsp;Ping Lu ,&nbsp;Yongsheng Yu ,&nbsp;Weiyu Gu ,&nbsp;Chunyu Li ,&nbsp;Yanqiu Lv ,&nbsp;Xinglin Qu ,&nbsp;Yuyang Zhang ,&nbsp;Qinglong Xu ,&nbsp;Shunfa Yao ,&nbsp;Xuan Chen ,&nbsp;Yi Jin","doi":"10.1016/j.theriogenology.2025.01.011","DOIUrl":"10.1016/j.theriogenology.2025.01.011","url":null,"abstract":"<div><div>Follicular fluid extracellular vesicles are beneficial for in vitro oocyte maturation and development; however, their effect on the expression profiles of oocyte microRNAs (miRNAs) and the roles of related miRNAs are unknown. In this study, we aimed to investigate miRNA expression in mature oocytes cultured in follicular fluid extracellular vesicles and the effect of miRNA-125a (miR-125a) on oocyte maturation. The expression profiles of the miRNAs were determined by microRNA sequencing, followed by target gene prediction analysis. We transfected miR-125a mimics and an miR-125a inhibitor to evaluate the effect of modulated miRNA-125a on cumulus expansion, oocyte maturation rate, changes in cytoplasmic maturation-related indicators, and changes in the expression of oocyte maturation-related, cumulus expansion-related, and predicted target genes. We found that miR-125a overexpression decreased the levels of cumulus expansion-related, oocyte maturation-related, and predicted target genes, adenosine deaminase RNA specific <em>(ADAR</em>), and lipid droplet number, and it increased the percentage of oocytes with abnormal cortical granule distribution. Inhibiting miR-125a increased the expression levels of oocyte maturation-related and target genes, number of lipid droplets, and endoplasmic reticulum function, and it decreased lipid droplet size. Mitochondrial membrane potential and reactive oxygen species levels were not significantly different between groups. In conclusion, our results suggest that extracellular vesicles may improve oocyte quality by modulating <em>ADAR</em> through regulating miR-125a.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 184-193"},"PeriodicalIF":2.4,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024770","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":"Strategic use of estrus intensity to combine timed artificial insemination and embryo transfer in large-scale cattle reproduction programs","authors":"Fábio Morotti ,&nbsp;Gustavo Martins Gomes dos Santos ,&nbsp;Katia Cristina Silva-Santos ,&nbsp;José Henrique Ayres Dias ,&nbsp;Marcelo Marcondes Seneda","doi":"10.1016/j.theriogenology.2025.01.009","DOIUrl":"10.1016/j.theriogenology.2025.01.009","url":null,"abstract":"<div><div>This study compared the reproductive performance of three different programs using conventional fixed-time artificial insemination (FTAI), fixed-time embryo transfer (FTET), and their combined use, considering estrus intensity as a criterion for the reproductive program. Brangus multiparous cows (n = 1.100), 40–50 days postpartum, 4–8 years old, and body condition scores 2.50 of 4.50 underwent a protocol for ovulation synchronization based on progesterone (P4) and estradiol on D0. On D10, one of three Programs I-III was implemented: control FTAI (n = 147 cows), control FTET (n = 617) with embryos from <em>in vitro</em> production (IVP) on D17, or FTAI + FTET (n = 336), where cows with low or no estrus expression were inseminated on D10, similar to Program I, while those with high-intensity expression received one embryo from IVP on D17, similar to Program II. Corpus luteum (CL) size and quality were assessed using B-mode and Doppler ultrasound on D17. The service/utilization rate was higher for FTAI and combined FTAI + FTET than for the conventional FTET. The program that used only FTAI resulted in higher pregnancy at 30 and 60 days, as well as lower pregnancy loss, compared to programs that used embryos. Furthermore, the FTAI + FTET program showed reduced pregnancy loss, compared to the FTET program. In the combined program, the CL was greater in those who received FTET alone than in those who received FTAI alone. A higher proportion of recipients with CL with a high luteal blood perfusion score was observed in the FTET group compared to the FTAI group. Satisfactory reproductive rates can be achieved using conventional FTAI or FTET programs. However, the combined program, associated with monitoring the intensity of estrus expression, is a promising strategy for allocating females with absent or low estrus expression to FTAI, and those with high estrus intensity to FTET. Recipients with high-intensity expression had higher CL quality and, when transferred, maintained satisfactory reproductive performance compared with conventional FTET.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 162-167"},"PeriodicalIF":2.4,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012175","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":"The occurrence of follicular cyst affects the embryonic developmental competences of buffalo oocytes under in vitro culture conditions","authors":"Pradeep Saini ,&nbsp;Sandeep Kumar ,&nbsp;Hitesh Jangra ,&nbsp;Anupama Solanki ,&nbsp;Rahul Verma ,&nbsp;Rajesh Kumar ,&nbsp;Ramakant Kaushik ,&nbsp;Kushal Grakh ,&nbsp;Gitesh Saini ,&nbsp;Yogesh Bangar ,&nbsp;Anand Kumar Pandey","doi":"10.1016/j.theriogenology.2025.01.007","DOIUrl":"10.1016/j.theriogenology.2025.01.007","url":null,"abstract":"<div><div>Cystic ovarian disease (COD) is a major cause of infertility in dairy cows. This study aimed to investigate the impact of follicular cysts on the <em>invitro</em> blastocyst developmental competence of oocytes and the relative gene expression of blastocysts developed from the subordinate follicles of ipsilateral (ovary with cyst), contralateral (ovary opposite to cyst), and normal ovaries of buffaloes. A total of 2059 ovaries were collected from slaughterhouse and classified into three categories based on the presence of follicular cysts: a) ipsilateral, b) contralateral, and c) control (absence of cysts). Oocytes of grades A, B, and C were used for <em>invitro</em> maturation, <em>invitro</em> fertilization<em>,</em> and <em>invitro</em> culture. The cleavage rates of the ipsilateral (91.54 %) and contralateral (95.71 %) categories were higher (P &lt; 0.01) than those of the control (76.82 %). Conversely, the blastocyst development rate was higher (P &lt; 0.01) in control (32.67 %) than in ipsilateral (8.46 %) and contralateral (7.14 %) groups. The mRNA expression levels of maturation (BMP15), steroidogenesis (STAR, CYP19), antioxidant (SOD2, HSPB1) and anti-apoptotic (BCL2) genes were decreased in arrested embryos derived from both cystic (ipsilateral and contralateral) and control category follicles when compared to blastocysts derived from their respective category follicles. Conversely, BAX expression increased in arrested embryos. Expression of SOD2 and BAX was downregulated in blastocysts from both ipsilateral and contralateral categories compared to controls. The presence of one or more follicular cysts in either ovary affected the developmental competence of oocytes derived from subordinate follicles. Therefore, the buffaloes with cysts in either ovary should be avoided when aspirating follicles for IVF.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 152-161"},"PeriodicalIF":2.4,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012194","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":"The effects of mitoquinol mesylate on post-thaw characteristics and fertilizing ability of rooster semen cryopreserved in the Beltsville medium","authors":"N. Asadzadeh ,&nbsp;R. Masoudi ,&nbsp;R. Nateghi ,&nbsp;N.D. Davachi ,&nbsp;H.J. Barfourooshi ,&nbsp;P.M. Bartlewski","doi":"10.1016/j.theriogenology.2025.01.008","DOIUrl":"10.1016/j.theriogenology.2025.01.008","url":null,"abstract":"<div><div>Cryopreservation of rooster semen is a reproductive technology carried out to boost genetic gain and productivity in commercial flocks of chicken. However, semen freezing significantly reduces the quality and fertilizing potential of spermatozoa. This study examined cryoprotective effects of the mitochondria-targeted antioxidant mitoquinol mesylate added to the freezing extender by assessing post-thaw characteristics of rooster sperm. Semen samples were diluted in the Beltsville extender supplemented with 0, 1, 10, 100 or 1000 nM of mitoquinol mesylate. Following the thawing of cryopreserved semen doses, we evaluated the following sperm parameters: motility and morphology, membrane integrity and mitochondrial function, acrosome integrity, apoptosis status, lipid peroxidation, DNA fragmentation, reactive oxygen species (ROS) accumulation, epigenetic patterns (DNA methylation and histone modifications), and fertilizing ability. Semen diluted in extenders containing 10 or 100 nM of mitoquinol mesylate significantly exceeded all other groups of frozen-thawed semen samples in sperm motility, average path velocity as well as membrane/acrosome integrity, mitochondrial function indices and DNA/histone modification. Moreover, the addition of 10 and 100 nM of mitoquinol mesylate significantly reduced lipid peroxidation, apoptosis rate, DNA fragmentation and ROS concentrations compared with all other dilutions and was associated with a higher (P ≤ 0.05) fertilization rate compared with a non-supplemented control group, albeit it was still significantly lower compared with that obtained using fresh semen. It can be, however, concluded that mitoquinol mesylate significantly improved an array of quality parameters and fertilizing potential of rooster semen, and hence can be recommended for use as a diluent additive in semen cryopreservation procedures employed in commercial poultry operations.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 114-120"},"PeriodicalIF":2.4,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012176","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":"Antagonistic interaction between miR-143 and KRAS gene regulating male mouse germ cell apoptosis","authors":"Yu Lu ,&nbsp;Shudong Niu ,&nbsp;Guisheng Zhang,&nbsp;Yanfeng Guo,&nbsp;Baotong Fu,&nbsp;Miaomiao Wang,&nbsp;Jianan Liu,&nbsp;Haili Zhang,&nbsp;Wen Lu,&nbsp;Ming Zhang","doi":"10.1016/j.theriogenology.2024.12.024","DOIUrl":"10.1016/j.theriogenology.2024.12.024","url":null,"abstract":"<div><div>Precisely regulated spermatocyte growth, differentiation, and apoptosis are crucial for sustainable male fertility. miR-143 has been demonstrated to regulate gene expression and cell apoptosis in various human cancers. However, the function of mmu-mir-143 (miR-143) in mammalian testes and its underlying mechanism remains unexplored. In this study, the expression of miR-143 was detected in C57BL/6 mice spermatocytes by in situ hybridization (ISH) and immunofluorescence (IF) co-staining and transfecting miR-143 inhibitor into GC-2 cells (mouse spermatogenic cells) shows that miR-143 inhibits cleaved Caspase 3 (CC3)-induced male germ cell death. The current study used IF co-staining of KI67 and γ-H2A.X in the testes of C57BL/6 mice at different developmental stages, revealing that active proliferation and apoptosis of spermatocytes occurred simultaneously in the testes at 14 day post-partum (dpp). <em>Kras</em> was predicted as a potential target of miR-143 in mice using of the online database TargetScan, verified by quantitative real-time PCR (qPCR), western blotting (WB), and Dual-luciferase reporter gene assay. Co-transfection of miR-143 inhibitor and <em>Kras</em> siRNA into GC-2 cells revealed an antagonistic correlation between miR-143 and <em>Kras</em> in regulating male germ cell death. Finally, miR-143 inhibitor and mimics were administered into the seminiferous tubule of 3-week-old C57BL/6 mice. The histomorphology, IF co-staining, and WB data indicated that the testes treated with the miR-143 inhibitor showed significantly aberrant phenotypes, including damaged seminiferous tubules, reduced spermatocyte quantity, and elevated levels of apoptosis. This study uncovered the mechanism by which miR-143 inhibits male germ cell apoptosis through the repression of <em>Kras/KRAS</em> levels and the inhibition of Caspase 3 activation, providing insight into the role of miRNA in spermatogenesis and the maintenance of male fertility.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 121-133"},"PeriodicalIF":2.4,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012245","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":"R-spondin1 plays an indispensable role in ovarian development of Qi River crucian carp (Carassius auratus) by regulating estrogen synthesis","authors":"Limin Wu ,&nbsp;Mengfan Wu ,&nbsp;Yongjing Li ,&nbsp;Qingqing Xin ,&nbsp;Yuchi Wang ,&nbsp;Xi Shi ,&nbsp;Xuejun Li","doi":"10.1016/j.theriogenology.2025.01.006","DOIUrl":"10.1016/j.theriogenology.2025.01.006","url":null,"abstract":"<div><div>R-spondin1 (Rspo1) is a member of the secreted furin-like domain-containing protein family, and it is recognized for its significance in mammalian ovarian development. However, its role in teleost ovarian development remains largely uninvestigated. The Qi River crucian carp (<em>Carassius auratus</em>) is a species capable of gynogenesis, and it encounters challenges of premature ovarian maturation in aquaculture settings. Previous research established the essential involvement of Rspo1 in oocyte growth in Qi River crucian carp, but the precise molecular mechanisms underlying its role remain poorly understood. In this study, we categorized the pre-spawning ovarian development process of premature Qi River crucian carp into five stages through meticulous examination of morphology and histology. Immunofluorescence analysis revealed colocalization of Rspo1 with Vasa protein in oogonia, primary growth stage, and cortical vacuolar stage oocytes, and it was also detected in somatic cells. After a 60-day period of RNA interference via injection of <em>Rspo1</em> double-stranded RNA into late-previtellogenesis stage ovaries, a substantial proportion of oocytes were arrested in the primary growth stage and exhibited a marked reduction in the expression of germ cell marker genes and an increase in apoptosis signaling. RNA-sequencing and real-time PCR analyses indicated a potential association between genes involved in hormone synthesis, lipid storage, and cell proliferation with ovary development in Qi River crucian carp. Furthermore, a significant decrease in levels of serum estrogens and vitellogenin was observed after <em>Rspo1</em> knockdown. Dual-fluorescence <em>in situ</em> hybridization analysis demonstrated co-expression of <em>Rspo1</em> with <em>cyp19a1a</em> in ovarian germ and surrounding somatic cells. Furthermore, results of a promoter assay indicated that <em>Rspo1</em> can dose-dependently activate <em>cyp19a1a</em> expression. Collectively, these findings suggest that Rspo1 plays a role in ovarian development and oocyte growth by modulating <em>cyp19a1a</em> expression and influencing estrogen synthesis. These results provide valuable insights into the molecular mechanisms underlying the involvement of Rspo1 in ovarian development in Qi River crucian carp.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 134-144"},"PeriodicalIF":2.4,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012174","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 antifreeze protein III on canine sperm cryopreservation","authors":"Abbas Farshad,&nbsp;Emilia Diel,&nbsp;Axel Wehrend","doi":"10.1016/j.theriogenology.2025.01.004","DOIUrl":"10.1016/j.theriogenology.2025.01.004","url":null,"abstract":"<div><div>Sperm cryopreservation is crucial in reproductive biotechnology; however, the longevity of frozen and thawed semen is limited by the deterioration of sperm cell integrity. This study aimed to examine the effects of adding antifreeze protein III (AFP III) to the diluent, using samples from eight healthy mature dogs. The ejaculates were divided into aliquots and diluted with a standard Tris-fructose-egg yolk extender containing AFP III at concentrations of 0, 0.75, 1.0, and 2.0 μg/ml. After thawing, the samples were analyzed for kinematic parameters, membrane Integrity, lipid peroxidation, viability, acrosome integrity, intracellular hydrogen peroxide, mitochondrial membrane potential and apoptotic metrics. The results show that while motility and velocity were not significantly different between the treated and control groups (p &gt; 0.05), the treated groups generally performed better. Specifically, the 0.75 and 1.0 μg/ml groups exhibited better movement compared to the 2.0 μg/ml group. Additionally, there was a significant difference (p &lt; 0.05) in membrane integrity between the control and treated groups, though no differences were observed among the treated groups. Significant differences (p &lt; 0.05) were also observed in viability and acrosome integrity, with the 0.75 and 1.0 μg/ml groups outperforming the control and 2.0 μg/ml groups. There were no significant variations (p &gt; 0.05) in phosphatidylserine translocation, lipid peroxidation, mitochondrial membrane potential, or hydrogen peroxide levels. However, the 0.75 and 1.0 μg/ml groups demonstrated superior effects compared to both the control and the 2.0 μg/ml groups. These results suggest that the addition of antifreeze proteins, specifically AFP III, markedly improves the protection of canine sperm during cryopreservation. This enhancement is evident in various parameters, underscoring the beneficial effects of AFP III in maintaining sperm quality.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 86-93"},"PeriodicalIF":2.4,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142972185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}