TheriogenologyPub Date : 2025-01-11DOI: 10.1016/j.theriogenology.2025.01.008
N. Asadzadeh , R. Masoudi , R. Nateghi , N.D. Davachi , H.J. Barfourooshi , P.M. Bartlewski
{"title":"The effects of mitoquinol mesylate on post-thaw characteristics and fertilizing ability of rooster semen cryopreserved in the Beltsville medium","authors":"N. Asadzadeh , R. Masoudi , R. Nateghi , N.D. Davachi , H.J. Barfourooshi , 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 , Shudong Niu , Guisheng Zhang, Yanfeng Guo, Baotong Fu, Miaomiao Wang, Jianan Liu, Haili Zhang, Wen Lu, 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}
TheriogenologyPub Date : 2025-01-11DOI: 10.1016/j.theriogenology.2025.01.006
Limin Wu , Mengfan Wu , Yongjing Li , Qingqing Xin , Yuchi Wang , Xi Shi , Xuejun Li
{"title":"R-spondin1 plays an indispensable role in ovarian development of Qi River crucian carp (Carassius auratus) by regulating estrogen synthesis","authors":"Limin Wu , Mengfan Wu , Yongjing Li , Qingqing Xin , Yuchi Wang , Xi Shi , 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}
TheriogenologyPub Date : 2025-01-07DOI: 10.1016/j.theriogenology.2025.01.004
Abbas Farshad, Emilia Diel, Axel Wehrend
{"title":"Influence of antifreeze protein III on canine sperm cryopreservation","authors":"Abbas Farshad, Emilia Diel, 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 > 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 < 0.05) in membrane integrity between the control and treated groups, though no differences were observed among the treated groups. Significant differences (p < 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 > 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}
TheriogenologyPub Date : 2025-01-07DOI: 10.1016/j.theriogenology.2025.01.005
Naru Zhou , Zongliang Liu , Zhenhu Shi , Lei Luo , Mengqing Xuan , Ruiqing Zhu , Kunlong Hu , Xinyue Zhu , Wenhuan Xu , Yunsheng Li , Zubing Cao , Yunhai Zhang
{"title":"PARL regulates porcine oocyte meiotic maturation by mediating mitochondrial activity","authors":"Naru Zhou , Zongliang Liu , Zhenhu Shi , Lei Luo , Mengqing Xuan , Ruiqing Zhu , Kunlong Hu , Xinyue Zhu , Wenhuan Xu , Yunsheng Li , Zubing Cao , Yunhai Zhang","doi":"10.1016/j.theriogenology.2025.01.005","DOIUrl":"10.1016/j.theriogenology.2025.01.005","url":null,"abstract":"<div><div>PARL is a rhomboid membrane protein that plays a crucial role in regulating the metabolism and maintaining the homeostasis of mitochondria which provide important energy and material reserves for oocyte maturation. However, the impact of PARL on oocyte maturation remains poorly understood. Here, we elucidated the pivotal role of PARL in oocyte maturation through its regulatory effects on mitochondrial activity. Specifically, our findings revealed that inhibiting PARL expression by interfering with RNA transcription in oocytes led to a substantial decrease in the rate of first polar body extrusion and early development of parthenogenetically activated embryos. Moreover, PARL deficiency disrupted mitochondrial distribution and activity, leading to the accumulation of ROS, abnormal distribution of CGs and actin, increased tubulin acetylation modification, disturbed spindle assembly and chromosome alignment, ultimately caused DNA damage in porcine oocytes at the metaphase II stage. Intriguingly, PARL deficiency did not cause occurrence of apoptosis in oocytes. Furthermore, our study highlighted that PARL deficiency caused the aberrant expression of genes associated with oocyte maturation, particularly those genes associated with mitochondrial function and DNA integrity. Collectively, these results demonstrate that the indispensable role of PARL in orchestrating porcine oocyte meiotic maturation though its modulation of mitochondrial activity.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 75-85"},"PeriodicalIF":2.4,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142972186","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}
TheriogenologyPub Date : 2025-01-06DOI: 10.1016/j.theriogenology.2025.01.003
Kexin Li , Xiaolei Wang , Liang Ma , Youshe Ren , Lei Shi
{"title":"The mechanism of Se in regulating the proliferation and apoptosis of sheep Leydig cells through the miR-200a/NRF2 pathway","authors":"Kexin Li , Xiaolei Wang , Liang Ma , Youshe Ren , Lei Shi","doi":"10.1016/j.theriogenology.2025.01.003","DOIUrl":"10.1016/j.theriogenology.2025.01.003","url":null,"abstract":"<div><div>This study aimed to investigate the mechanism by which Se in regulates the proliferation and apoptosis of sheep Leydig cells via the miR-200a/NRF pathway. The cells were isolated and purified from the testes of 8-month-old sheep via a Percoll density gradient. After the cells were treated with different concentrations of Se (0, 2.0, 4.0, 6.0, and 8.0 μmol/L of Se) for 18 h, the miR-200a levels was detected. MiR-200a mimics and inhibitors were transfected into the cells, resulting in five groups (control, NC mimics, miR-200a mimics, NC inhibitor and miR-200a inhibitor). Cell viability and antioxidant status were measured via CCK8 and antioxidant assays, respectively. The abundances of pro-apoptotic (<em>BAX</em>, <em>CASPASE 3</em> and <em>CASPASE 8</em>), cell cycle (<em>P21</em>, <em>P27</em> and <em>CDK1</em>), and NRF2-related (<em>NRF2</em>, <em>HO-1</em>, <em>NQO1</em> and <em>KEAP1</em>) genes were detected by real-time PCR and Western blot analysis.</div><div>The results revealed that miR-200a mimics group presented greater (<em>P</em> < 0.05) abundances of <em>NRF2</em>, <em>HO-1</em> and <em>NQO1</em> mRNA transcripts and proteins. Compared with those both in the NC mimics and the miR-200a inhibitor groups, the activities of GSH-Px and SOD, as well as cell viability in the miR-200a mimics group were significantly greater (<em>P</em> < 0.05). In contrast, the ROS levels, MDA content and abundances of <em>KEAP1</em>, <em>P21</em>, <em>P27</em> and apoptosis-related genes mRNA transcripts and proteins were decreased (<em>P</em> < 0.05). The highest (<em>P</em> < 0.05) miR-200a expression level was detected in the Se<sub>6.0</sub> group. Compared with that in the Se (6.0 μmol/L) group, cell viability in the Se + miR-200a inhibitor group was lower (<em>P</em> < 0.05). The abundances of <em>NRF2</em>, <em>HO-1</em> and <em>NQO1</em> in the Se + miR-200a inhibitor group were lower (<em>P</em> < 0.05) than those in the Se (6.0 μmol/L) group but greater (<em>P</em> < 0.05) than those in the inhibitor group, while <em>KEAP1</em> displayed the opposite trend (<em>P</em> < 0.05).</div><div>These results indicate that Se can activate the NRF2 antioxidant signaling pathway to regulate the proliferation and apoptosis of sheep Leydig cells and that miR-200a plays a vital role in this process. The regulatory effect of Se on male reproduction and spermatogenesis may be related to the number of Leydig cells. This study aimed to provide experimental data for Se regulation of spermatogenesis.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 103-113"},"PeriodicalIF":2.4,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142984804","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}
TheriogenologyPub Date : 2025-01-06DOI: 10.1016/j.theriogenology.2025.01.001
Cecilia Constantino Rocha , Ana Beatriz Montevecchio , Alexandra Bennett , Abdul Waheed , Mackenzie Mazziotta , Tatiane S. Maia , McKenzie Lane-Jackson Haimon , Quinn A. Hoorn , Masroor Sagheer , Camila J. Cuellar , Oscar Alejandro Ojeda-Rojas , Rebecca Lynn Krisher , Marcello Rubessa , Ky G. Pohler , Peter J. Hansen , Philipe Moriel , Ricardo C. Chebel , Mario Binelli
{"title":"Relationships between activity monitoring device data and ovarian, uterine, hormonal, and pregnancy variables in beef cows","authors":"Cecilia Constantino Rocha , Ana Beatriz Montevecchio , Alexandra Bennett , Abdul Waheed , Mackenzie Mazziotta , Tatiane S. Maia , McKenzie Lane-Jackson Haimon , Quinn A. Hoorn , Masroor Sagheer , Camila J. Cuellar , Oscar Alejandro Ojeda-Rojas , Rebecca Lynn Krisher , Marcello Rubessa , Ky G. Pohler , Peter J. Hansen , Philipe Moriel , Ricardo C. Chebel , Mario Binelli","doi":"10.1016/j.theriogenology.2025.01.001","DOIUrl":"10.1016/j.theriogenology.2025.01.001","url":null,"abstract":"<div><div>Implementing accelerometer technologies in beef operations is an alternative to increase precision in estrous detection. We hypothesized that (1) the accelerometer algorithm has similar accuracy in detecting behavioral estrus as does visual observation of pressure-sensitive sensors (estrus patches) in grazing beef cows; (2) variables measured by the accelerometer, such as estrus intensity, are associated with hormonal, ovarian, and uterine variables monitored before, during, and after estrus; and (3) the accelerometer variables are associated with the probability of pregnancy in grazing beef cows submitted to embryo transfer (ET). Fifty cows were fitted with accelerometer and patches to detect estrus after a synchronization protocol in eight subsequent rounds. For each round, only cows that showed estrus (day 0; D0) received ET. Follicular diameter, endometrial thickness, corpus luteum (CL) area, and estradiol (E2) and progesterone (P4) concentrations were measured during proestrus, estrus, and early diestrus. On D7, ET was performed. Pregnancies were diagnosed on D46 and cows recovered for 35D before a new replicate. Patches had a greater accuracy (98 % vs. 91 %) of detection of behavioral estrus than accelerometer algorithm. Cows with lower estrus intensity in the accelerometer had greater follicular diameter on D0 (P = 0.022), CL area on D4 and D7 (P = 0.05), endometrial thickness on D-1 (P = 0.10), and reduced E2 concentrations on D-1 (P = 0.0032). The accelerometer variables did not predict accurately the probability of pregnancy/ET. In conclusion, visual observation of patches was more accurate in detecting estrus than the accelerometer algorithm and most of the associations between accelerometers and physiological variables were for characteristics measured at proestrus.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 64-74"},"PeriodicalIF":2.4,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142967048","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}
TheriogenologyPub Date : 2025-01-06DOI: 10.1016/j.theriogenology.2025.01.002
García-Barcelo Pedro , Romero Angel , Rodríguez-Piñón Marcelo , Cerecetto Hugo
{"title":"In vitro study of carbetocin, an oxytocin receptor agonist, and 4-phenylfuroxan-3-carbonitrile, a NO-releasing agent, as cervical dilatators in sheep","authors":"García-Barcelo Pedro , Romero Angel , Rodríguez-Piñón Marcelo , Cerecetto Hugo","doi":"10.1016/j.theriogenology.2025.01.002","DOIUrl":"10.1016/j.theriogenology.2025.01.002","url":null,"abstract":"<div><div>The aim was to study the effect of 4-phenylfuroxan-3-carbonitrile (Fx), a NO-releasing agent, and carbetocin, an oxytocin receptor agonist, on matrix metalloproteinases-2 (MMP-2) activity and PGE2 production in cervix from cycling sheep. Cervical explants were incubated during 12 h with MEM supplemented with increasing concentrations of Fx in DMSO (2 %) (0 to 300 μg/mL) with Cb (100 ng/mL) (Experiment 1, n = 15) and DMSO (2 %), DMSO + Cb (100 ng/mL) or DMSO + Fx (30 μg/mL) (Experiment 2, n = 10), and their respective controls. In the supernatants, activated (A) and latent (L) MMP-2 activities were determined by a SDS-PAGE zymography, PGE2 concentration by immunoassay and NO production indirectly as nitrites by spectrophotometry. Data were analyzed by ANOVA. The Cb treatment increase the A MMP-2 activity in DMSO (Experiment 1 at follicular phase and Experiment 2) or alone (Experiment 2) and increase the L MMP-2 activity (Experiments 1 and 2) (P < 0.02). The DMSO treatment also increase the L MMP2 activity (Experiment 2) (P < 0.0001). Treatment with Fx + DMSO increased the concentration of accumulated nitrites in the supernatant (P < 0.0001) (Experiment 1), but did not affect or decrease the activity of A and L MMP-2 (P < 0.04) (Experiments 1 and 2). The PGE2 concentration trend to increase with Cb treatment (P = 0.0614) and decrease with Fx+DMSO treatment (P < 0.0001) (Experiment 2). In conclusion, Cb and/or DMSO treatment of cervical explants increase the MMP-2 activity through PGE2-independent mechanisms, but Fx in DMSO fail in this, suggesting that the pre-treatment with Cb and/or DMSO could be used to increase cervical dilation in ewes.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 168-174"},"PeriodicalIF":2.4,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024767","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}
TheriogenologyPub Date : 2025-01-02DOI: 10.1016/j.theriogenology.2024.12.031
João Paulo N. Andrade , Victor E. Gomez-León , Guilherme Madureira , Luma C. Sartori , Gustavo F. Grillo , Rafael R. Domingues , Meliton Fosado , Rodrigo V. Sala , Milo C. Wiltbank
{"title":"ReBreed21-ET: Evaluation of a rapid resynchronization program that allows timed embryo transfer every 21 days","authors":"João Paulo N. Andrade , Victor E. Gomez-León , Guilherme Madureira , Luma C. Sartori , Gustavo F. Grillo , Rafael R. Domingues , Meliton Fosado , Rodrigo V. Sala , Milo C. Wiltbank","doi":"10.1016/j.theriogenology.2024.12.031","DOIUrl":"10.1016/j.theriogenology.2024.12.031","url":null,"abstract":"<div><div>This study evaluated the efficiency of a rapid resynchronization of ovulation program to allow timed embryo transfer (TET) every 21d in heifer embryo recipients. Holstein heifers (n = 510) had synchronized ovulation using a modified 5d CoSynch program for a TET (D7) after induced ovulation (D0). After TET, heifers were blocked by number of previous TETs and randomized into one of two resynchronization of ovulation programs: Resynch28 (n = 279), a traditional resynch program for TET 35d after previous TET; or ReBreed21-ET, a program designed to allow TET 21d after previous TET. Once assigned into one of the two programs, heifers were kept in the same program during a 105-d TET period. In Resynch28, heifers received an intravaginal progesterone (P4) insert on D28, on D33 the P4 was removed, and pregnancy diagnosis was performed using rectal ultrasound to determine embryonic heartbeat. Nonpregnant heifers received a PGF2α treatment with a later GnRH treatment on D35 for a potential TET on D42 (35d after previous TET). Heifers in ReBreed21-TET received an intravaginal P4 insert on D14, on D19 the P4 was removed, and on D21 a GnRH treatment was given to synchronize a new ovulation. On D28, pregnancy diagnosis was performed using transrectal ultrasound to detect an embryonic heartbeat and nonpregnant heifers that had a corpus luteum (CL) ≥18 mm in diameter received a TET (21d after the previous TET). Pregnancy per ET (P/ET) from the first TET was greater for heifers in ReBreed21-ET (52 %) than Resynch28 (39.4 %). In contrast, the subsequent TET (second and later) had similar P/ET for ReBreed21-ET (40.4 %) and Resynch28 (40.8 %). The overall pregnancy loss from D28 to D63 did not differ between programs (ReBreed21-ET [18.5 %] and Resynch28 (16.3 %]). Nevertheless, there were fewer pregnancy losses from D28 to D33 for Resynch28 (3.5 %) than ReBreed21-ET (10.1 %), while from D33 to D47, there was greater pregnancy loss for Resynch28 (10.1 %) than ReBreed21-ET (4.9 %). Time to pregnancy was approximately 8d earlier for ReBreed21-ET (45 ± 3d) than Resynch28 (53 ± 3d). Overall cumulative pregnancies at the end of a 105-d TET season were greater for ReBreed21-ET (75.2 %) than Resynch28 (64 %). Thus, the ReBreed21-ET program can improve the efficiency of TET programs.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 145-151"},"PeriodicalIF":2.4,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012172","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}
TheriogenologyPub Date : 2025-01-02DOI: 10.1016/j.theriogenology.2024.12.022
Annie L. Whitty , Karen L. Kind , Kylie R. Dunning , Nicole O. McPherson , Mark B. Nottle
{"title":"GM-CSF treatment of frozen bovine sperm improves function, fertilization, and subsequent embryo development","authors":"Annie L. Whitty , Karen L. Kind , Kylie R. Dunning , Nicole O. McPherson , Mark B. Nottle","doi":"10.1016/j.theriogenology.2024.12.022","DOIUrl":"10.1016/j.theriogenology.2024.12.022","url":null,"abstract":"<div><div>In vitro embryo production (IVP) is used in the cattle industry to increase the rate of genetic gain. IVP uses semen that has been frozen and thawed, a process that renders sperm less viable than sperm from fresh semen. Granulocyte macrophage colony stimulating factor (GM-CSF) is present in bovine seminal plasma, while its receptor is present on bovine sperm. The present study aimed to determine if GM-CSF could improve the function and quality of frozen-thawed bovine sperm and embryo development following in vitro fertilization. Thawed bovine sperm (n = 3 bulls with 3 replicates per bull) was incubated with 0, 0.1, 1, 2 or 10 ng/ml of recombinant bovine GM-CSF in buffered wash media for 45 min and assessed for motility, glucose uptake, mitochondrial activity, intracellular calcium, capacitation, DNA integrity, and in vitro embryo development. The addition of 1, 2, and 10 ng/ml GM-CSF increased total motility (P = 0.02, P = 0.007, P = 0.01), progressive (P = 0.02, P = 0.03, P = 0.01), and rapid motility (P = 0.01, P = 0.01, P = 0.01), while 10 ng/ml increased glucose uptake (P = 0.003), and 1, 2, and 10 ng/ml increased capacitation (P = 0.003, P = 0.001, P = 0.0003). There was no difference between groups for mitochondrial activity, intracellular calcium, or DNA integrity. GM-CSF treatment of sperm prior to in vitro insemination increased fertilization rate (P = 0.01), hatching blastocyst rate (P = 0.05), and blastocyst inner cell mass cell number (P = 0.03) compared with control. In conclusion, GM-CSF treatment of frozen-thawed bovine sperm improves sperm function and quality resulting in increased fertilization capacity and subsequent embryo development, suggesting it may improve cattle IVP efficiencies.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 46-55"},"PeriodicalIF":2.4,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143130055","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}