Theriogenology最新文献_第2页

High out-of-season fertility in progesterone-treated goats joined with sexually active bucks and artificially inseminated with fresh diluted semen

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-02-05 DOI: 10.1016/j.theriogenology.2025.02.003

J.A. Delgadillo , D. López-Magaña , G. Duarte , N. López-Magaña , P. Chemineau

In seasonal anestrous goats, artificial insemination (AI) is typically preceded by hormonal treatment. This study examined whether high fertility can be achieved after AI with fresh diluted semen in progesterone (P4)-treated goats exposed to sexually active (SA) bucks. In Experiment 1, two groups of goats (n = 30 each) were joined with SA bucks (n = 2 per group) fitted with abdominal aprons. Prior to teasing, the P4-group received an intramuscular injection of 25 mg of progesterone diluted in 2 mL of olive oil, while the control group received olive oil alone. AI was performed 12 h after estrus onset within five days after buck introduction, using semen packed in 0.2 mL straws containing 80 × 10⁶ spermatozoa. The kidding rate was higher in the P4-treated group (24/30, 80 %) compared to the control group (8/30, 27 %; P < 0.001). In Experiment 2, two groups (n = 30 each) were exposed to SA bucks (n = 2 per group). The control group was inseminated 12 h after estrus onset detected between six- and nine-days post-introduction, while the P4-group was inseminated within five days. Kidding rates were similar between the P4-group (26/30, 86 %) and the control group (23/30, 77 %; P > 0.05). These results indicate that P4-treated goats achieve high fertility when inseminated during the first estrus induced by males, while control goats achieve high fertility when inseminated during the second estrus.

{"title":"High out-of-season fertility in progesterone-treated goats joined with sexually active bucks and artificially inseminated with fresh diluted semen","authors":"J.A. Delgadillo , D. López-Magaña , G. Duarte , N. López-Magaña , P. Chemineau","doi":"10.1016/j.theriogenology.2025.02.003","DOIUrl":"10.1016/j.theriogenology.2025.02.003","url":null,"abstract":"<div><div>In seasonal anestrous goats, artificial insemination (AI) is typically preceded by hormonal treatment. This study examined whether high fertility can be achieved after AI with fresh diluted semen in progesterone (P4)-treated goats exposed to sexually active (SA) bucks. In Experiment 1, two groups of goats (n = 30 each) were joined with SA bucks (n = 2 per group) fitted with abdominal aprons. Prior to teasing, the P4-group received an intramuscular injection of 25 mg of progesterone diluted in 2 mL of olive oil, while the control group received olive oil alone. AI was performed 12 h after estrus onset within five days after buck introduction, using semen packed in 0.2 mL straws containing 80 × 10⁶ spermatozoa. The kidding rate was higher in the P4-treated group (24/30, 80 %) compared to the control group (8/30, 27 %; P < 0.001). In Experiment 2, two groups (n = 30 each) were exposed to SA bucks (n = 2 per group). The control group was inseminated 12 h after estrus onset detected between six- and nine-days post-introduction, while the P4-group was inseminated within five days. Kidding rates were similar between the P4-group (26/30, 86 %) and the control group (23/30, 77 %; P > 0.05). These results indicate that P4-treated goats achieve high fertility when inseminated during the first estrus induced by males, while control goats achieve high fertility when inseminated during the second estrus.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 114-119"},"PeriodicalIF":2.4,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143349272","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}

引用次数: 0

Effect of bovine oviductal epithelial cell lysate on the developmental competence and quality of bovine in vitro fertilized embryos

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-02-03 DOI: 10.1016/j.theriogenology.2025.02.002

Funa Ota , Hayato Minowa , Rina Miura , Tetsuma Murase , Tokunori Yamamoto , Takehiro Himaki

In vitro fertilization (IVF) technology for embryo production has been applied in basic research, animal husbandry and medicine. However, the developmental efficiency and quality of embryos produced by IVF are inferior to those produced in vivo. In this study, we investigated the effects of supplementing bovine oviductal epithelial cells (BOEC) lysate during the in vitro culture period on the developmental competence and quality of bovine embryos. IVF embryos were cultured for 4 days post-IVF in medium supplemented with 10 % BOEC lysate at various concentrations (1.0 × 10⁵, 2.0 × 10⁵, and 4.0 × 10⁵ cells/mL) or 10 % PBS (−), which was used to adjust the lysate concentration (control). BOEC lysate at 2.0 × 10⁵ cells/mL significantly increased the blastocyst formation rate compared to that in the control group. Blastocysts from BOEC lysate supplemented groups showed significantly lower apoptosis rate than that in the control group. The ratio of inner cell mass cell number in blastocysts was significantly higher in all BOEC lysate supplemented groups than in the control group. The survival rate after vitrification/thawing was improved in the 1.0 × 10⁵ and 2.0 × 10⁵ cells/mL BOEC lysate supplemented groups. In addition, gene expression analysis of blastocysts showed that 2.0 × 10⁵ cells/mL of BOEC lysate supplementation significantly enhanced the expression of anti-apoptotic genes (BCL2 and BIRC5), antioxidant-related genes (GPX1 and SOD2), and cell differentiation-related genes (SOX2 and OCT4). In conclusion, supplementation with 2.0 × 10⁵ cells/mL BOEC lysate during early in vitro culture improved the developmental competence and quality of bovine IVF embryos.

{"title":"Effect of bovine oviductal epithelial cell lysate on the developmental competence and quality of bovine in vitro fertilized embryos","authors":"Funa Ota , Hayato Minowa , Rina Miura , Tetsuma Murase , Tokunori Yamamoto , Takehiro Himaki","doi":"10.1016/j.theriogenology.2025.02.002","DOIUrl":"10.1016/j.theriogenology.2025.02.002","url":null,"abstract":"<div><div><em>In vitro</em> fertilization (IVF) technology for embryo production has been applied in basic research, animal husbandry and medicine. However, the developmental efficiency and quality of embryos produced by IVF are inferior to those produced <em>in vivo</em>. In this study, we investigated the effects of supplementing bovine oviductal epithelial cells (BOEC) lysate during the <em>in vitro</em> culture period on the developmental competence and quality of bovine embryos. IVF embryos were cultured for 4 days post-IVF in medium supplemented with 10 % BOEC lysate at various concentrations (1.0 × 10<sup>5</sup>, 2.0 × 10<sup>5</sup>, and 4.0 × 10<sup>5</sup> cells/mL) or 10 % PBS (−), which was used to adjust the lysate concentration (control). BOEC lysate at 2.0 × 10<sup>5</sup> cells/mL significantly increased the blastocyst formation rate compared to that in the control group. Blastocysts from BOEC lysate supplemented groups showed significantly lower apoptosis rate than that in the control group. The ratio of inner cell mass cell number in blastocysts was significantly higher in all BOEC lysate supplemented groups than in the control group. The survival rate after vitrification/thawing was improved in the 1.0 × 10<sup>5</sup> and 2.0 × 10<sup>5</sup> cells/mL BOEC lysate supplemented groups. In addition, gene expression analysis of blastocysts showed that 2.0 × 10<sup>5</sup> cells/mL of BOEC lysate supplementation significantly enhanced the expression of anti-apoptotic genes (<em>BCL2</em> and <em>BIRC5</em>), antioxidant-related genes (<em>GPX1</em> and <em>SOD2</em>), and cell differentiation-related genes (<em>SOX2</em> and <em>OCT4</em>). In conclusion, supplementation with 2.0 × 10<sup>5</sup> cells/mL BOEC lysate during early <em>in vitro</em> culture improved the developmental competence and quality of bovine IVF embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 96-104"},"PeriodicalIF":2.4,"publicationDate":"2025-02-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143349365","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}

引用次数: 0

Developmental stage specific effect of Mito-TEMPO on the expression outline of antioxidant genes, ROS balance and cryo-resilience of bovine IVP embryos

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-02-02 DOI: 10.1016/j.theriogenology.2025.01.030

M. Schreiber , N. Ghanem , M. Rahimi , H. Habermann , E. Tholen , M. Hoelker , E. Held-Hoelker

In vitro culture impairs mitochondrial metabolism of IVP derived bovine embryos resulting in accumulation of reactive oxygen species. Recently, the antioxidant Mito-TEMPO has attracted attention due to its capability to accumulate within mitochondria. In order to investigate the potential of Mito-TEMPO to improve quality of IVP derived embryos, this study analyzed the developmental stage specific effect of Mito-TEMPO on developmental capacity, ROS balance, expression outline of antioxidative genes and cryo-resilience of blastocysts. In three subsequent experiments Mito-TEMPO (1 μM) was added either to the maturation medium (MTM), the culture medium (MTC) or to both the maturation medium and the culture medium (MTMC). Concerning cleavage- and blastocyst rates, no effect of Mito-TEMPO supplementation could be detected, although MTM and/or MTC groups revealed significantly (p < 0.05) lower levels of ROS. Expression outline of antioxidative genes under study was not affected in MTM group, whereas down-regulation of the proapoptotic gene BAX was observed in MTM as well as MTMC groups. Moreover, Mito-TEMPO significantly affected expression outline of genes with antioxidative functions within mitochondria (SOD2, GPX1, GSTA4) in MTC and/or MTMC groups and peroxisomes (CAT) in MTMC group. In contrast, expression of genes acting predominately outside mitochondria (NFE2L2 and PRDX1) was not affected. Of high impact, the present study revealed for the first time greatly improved reexpansion and hatching rates of bovine vitrified-warmed embryos as a consequence of supplementation of Mito-TEMPO to culture media. Collectively, the present study successfully proved that Mito-TEMPO alleviates negative effects of the in vitro culture environment in bovine pre-implantation embryos.

{"title":"Developmental stage specific effect of Mito-TEMPO on the expression outline of antioxidant genes, ROS balance and cryo-resilience of bovine IVP embryos","authors":"M. Schreiber , N. Ghanem , M. Rahimi , H. Habermann , E. Tholen , M. Hoelker , E. Held-Hoelker","doi":"10.1016/j.theriogenology.2025.01.030","DOIUrl":"10.1016/j.theriogenology.2025.01.030","url":null,"abstract":"<div><div>In vitro culture impairs mitochondrial metabolism of IVP derived bovine embryos resulting in accumulation of reactive oxygen species. Recently, the antioxidant Mito-TEMPO has attracted attention due to its capability to accumulate within mitochondria. In order to investigate the potential of Mito-TEMPO to improve quality of IVP derived embryos, this study analyzed the developmental stage specific effect of Mito-TEMPO on developmental capacity, ROS balance, expression outline of antioxidative genes and cryo-resilience of blastocysts. In three subsequent experiments Mito-TEMPO (1 μM) was added either to the maturation medium (MTM), the culture medium (MTC) or to both the maturation medium and the culture medium (MTMC). Concerning cleavage- and blastocyst rates, no effect of Mito-TEMPO supplementation could be detected, although MTM and/or MTC groups revealed significantly (p < 0.05) lower levels of ROS. Expression outline of antioxidative genes under study was not affected in MTM group, whereas down-regulation of the proapoptotic gene <em>BAX</em> was observed in MTM as well as MTMC groups. Moreover, Mito-TEMPO significantly affected expression outline of genes with antioxidative functions within mitochondria (<em>SOD2, GPX1, GSTA4</em>) in MTC and/or MTMC groups and peroxisomes (<em>CAT</em>) in MTMC group. In contrast, expression of genes acting predominately outside mitochondria (<em>NFE2L2</em> and <em>PRDX1</em>) was not affected. Of high impact, the present study revealed for the first time greatly improved reexpansion and hatching rates of bovine vitrified-warmed embryos as a consequence of supplementation of Mito-TEMPO to culture media. Collectively, the present study successfully proved that Mito-TEMPO alleviates negative effects of the in vitro culture environment in bovine pre-implantation embryos.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 105-113"},"PeriodicalIF":2.4,"publicationDate":"2025-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143349366","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}

引用次数: 0

Characteristics of porcine oocyte-cumulus complexes derived from various sizes of antral follicles and classified by brilliant cresyl blue staining, and developmental competence of the oocytes

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-02-02 DOI: 10.1016/j.theriogenology.2025.02.001

Phong Ngoc Van, Son Quang Do, Wanniarachchige Tharindu Lakshitha Fonseka , Takuya Wakai, Hiroaki Funahashi

The present study sought to determine the characteristics of porcine oocyte-cumulus complexes (OCCs) derived from very small and small antral follicles (with less than 1 mm and 1–3 mm in diameter, respectively; VSF and SF) in comparison with controls from medium ones (with 3–6 mm in diameter; MF). Additionally, the present study examined the utility of brilliant cresyl blue (BCB) staining for assessing these OCCs. The incidence of BCB- oocytes in VSF- and SF-derived OCCs was higher than that in MF-derived OCCs. Although the meiotic and developmental competences of BCB+ oocytes from MF were superior to those from VSF and SF, blastocysts were successfully obtained from BCB+ oocytes even derived from VSF. The mean numbers of both total and viable cumulus cells surrounding an oocyte were significantly affected not only by the origin of the OCCs, but also by the BCB status of the oocytes (largest in MF-derived OCCs containing BCB+ oocytes). Although the outer and inner diameters of zona pellucida were affected by the origin of OCCs and the BCB status of oocytes (largest in MF-derived oocytes), the ooplasmic diameter of BCB+ oocytes did not differ among those derived from VSF, SF, and MF. Regardless of the BCB status, the transcriptional levels of G6PD and TKT in cumulus cells decreased during follicular development from VSF to MF, whereas the RPIA mRNA level in cumulus cells of MF-derived BCB+ OCCs was lower than in the others. These results underscore the utility of BCB staining for selecting MF-, SF-, and even VSF-derived OCCs containing oocytes with relatively higher meiotic and developmental competences, as well as the importance of having a sufficient number of healthy cumulus cells expressing genes related to the pentose phosphate pathway at lower levels.

{"title":"Characteristics of porcine oocyte-cumulus complexes derived from various sizes of antral follicles and classified by brilliant cresyl blue staining, and developmental competence of the oocytes","authors":"Phong Ngoc Van, Son Quang Do, Wanniarachchige Tharindu Lakshitha Fonseka , Takuya Wakai, Hiroaki Funahashi","doi":"10.1016/j.theriogenology.2025.02.001","DOIUrl":"10.1016/j.theriogenology.2025.02.001","url":null,"abstract":"<div><div>The present study sought to determine the characteristics of porcine oocyte-cumulus complexes (OCCs) derived from very small and small antral follicles (with less than 1 mm and 1–3 mm in diameter, respectively; VSF and SF) in comparison with controls from medium ones (with 3–6 mm in diameter; MF). Additionally, the present study examined the utility of brilliant cresyl blue (BCB) staining for assessing these OCCs. The incidence of BCB- oocytes in VSF- and SF-derived OCCs was higher than that in MF-derived OCCs. Although the meiotic and developmental competences of BCB+ oocytes from MF were superior to those from VSF and SF, blastocysts were successfully obtained from BCB+ oocytes even derived from VSF. The mean numbers of both total and viable cumulus cells surrounding an oocyte were significantly affected not only by the origin of the OCCs, but also by the BCB status of the oocytes (largest in MF-derived OCCs containing BCB+ oocytes). Although the outer and inner diameters of zona pellucida were affected by the origin of OCCs and the BCB status of oocytes (largest in MF-derived oocytes), the ooplasmic diameter of BCB+ oocytes did not differ among those derived from VSF, SF, and MF. Regardless of the BCB status, the transcriptional levels of <em>G6PD</em> and <em>TKT</em> in cumulus cells decreased during follicular development from VSF to MF, whereas the <em>RPIA</em> mRNA level in cumulus cells of MF-derived BCB+ OCCs was lower than in the others. These results underscore the utility of BCB staining for selecting MF-, SF-, and even VSF-derived OCCs containing oocytes with relatively higher meiotic and developmental competences, as well as the importance of having a sufficient number of healthy cumulus cells expressing genes related to the pentose phosphate pathway at lower levels.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 74-81"},"PeriodicalIF":2.4,"publicationDate":"2025-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143298027","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}

引用次数: 0

Deciphering the microbiome, lipopolysaccharides, and metabolome interplay: Unveiling putrescine's mechanism for enhancing sperm quality in heat-stressed boars

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-01-31 DOI: 10.1016/j.theriogenology.2025.01.027

Chenglong Yu , Hao Li , Lun Hua , Lianqiang Che , Bin Feng , Zhengfeng Fang , Shengyu Xu , Yong Zhuo , Jian Li , De Wu , Junjie Zhang , Yan Lin

Global warming has added to concerns regarding declining male fertility due to high temperatures. As a metabolite of arginine, putrescine improves gut health and promotes testicular development in boar; however, its action in improving semen quality under heat stress is unknown. Therefore, we assessed the effect of putrescine on the semen quality of boars in a heat stress model. Results showed that putrescine ameliorated the heat stress-induced decline in semen quality and testosterone levels in boars, confirmed by sperm viability, immobility rate, and apoptosis levels. Fecal microbial 16S rRNA sequencing showed that heat stress induces intestinal microecological dysregulation triggering an increase in the serum lipopolysaccharide (LPS) levels and reducing boar semen quality. A negative correlation between the Lachnospiraceae_XPB1014_group and LPS-binding protein (LBP) levels was observed. The Lachnospiraceae_XPB1014_group was reduced significantly under heat stress, and its relative abundance significantly increased after putrescine diet, which reduced both LPS and LBP in the serum of heat-stressed boars. Heat stress also affected plasma amino acid metabolism, and the regulation of plasma metabolism by putrescine can be attributed to its effects on LPS and the LBP owing to the significantly correlation of both with multiple plasma differential metabolites. Putrescine is thus considered to inhibit the increased serum LPS by acting on intestinal microorganisms, particularly by increasing the relative abundance of the Lachnospiraceae_XPB1014_group, and further modulate plasma amino acid metabolism to improve the semen quality in heat-stressed boars.

{"title":"Deciphering the microbiome, lipopolysaccharides, and metabolome interplay: Unveiling putrescine's mechanism for enhancing sperm quality in heat-stressed boars","authors":"Chenglong Yu , Hao Li , Lun Hua , Lianqiang Che , Bin Feng , Zhengfeng Fang , Shengyu Xu , Yong Zhuo , Jian Li , De Wu , Junjie Zhang , Yan Lin","doi":"10.1016/j.theriogenology.2025.01.027","DOIUrl":"10.1016/j.theriogenology.2025.01.027","url":null,"abstract":"<div><div>Global warming has added to concerns regarding declining male fertility due to high temperatures. As a metabolite of arginine, putrescine improves gut health and promotes testicular development in boar; however, its action in improving semen quality under heat stress is unknown. Therefore, we assessed the effect of putrescine on the semen quality of boars in a heat stress model. Results showed that putrescine ameliorated the heat stress-induced decline in semen quality and testosterone levels in boars, confirmed by sperm viability, immobility rate, and apoptosis levels. Fecal microbial 16S rRNA sequencing showed that heat stress induces intestinal microecological dysregulation triggering an increase in the serum lipopolysaccharide (LPS) levels and reducing boar semen quality. A negative correlation between the <em>Lachnospiraceae_XPB1014_group</em> and LPS-binding protein (LBP) levels was observed. The <em>Lachnospiraceae_XPB1014_group</em> was reduced significantly under heat stress, and its relative abundance significantly increased after putrescine diet, which reduced both LPS and LBP in the serum of heat-stressed boars. Heat stress also affected plasma amino acid metabolism, and the regulation of plasma metabolism by putrescine can be attributed to its effects on LPS and the LBP owing to the significantly correlation of both with multiple plasma differential metabolites. Putrescine is thus considered to inhibit the increased serum LPS by acting on intestinal microorganisms, particularly by increasing the relative abundance of the <em>Lachnospiraceae_XPB1014_group</em>, and further modulate plasma amino acid metabolism to improve the semen quality in heat-stressed boars.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 60-73"},"PeriodicalIF":2.4,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143298630","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}

引用次数: 0

Follicle mural granulosa cells encapsulated in sodium alginate gel improve developmental competence of porcine oocytes of in vitro maturation

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-01-31 DOI: 10.1016/j.theriogenology.2025.01.029

Qi Zhang, Yongfeng Zhou, Ran Ding, Qi Li, Xinglan An, Sheng Zhang, Ziyi Li

The maturation of oocytes has an important impact on the subsequent development of the embryo. However, during the in vitro maturation (IVM) of oocytes, oocytes are separated from the follicular environment, resulting in a low maturation rate of oocytes in vitro. In order to improve maturation rate of IVM of porcine oocytes, this study was conducted to investigate using sodium alginate (SA) to encapsulate porcine mural granulosa cells (MGs) to develop an SA three-dimensional (3D) co-culture system for IVM of porcine oocytes. And, gene expression, reactive oxygen species (ROS), ATP level, mitochondrial membrane potential, parthenogenetic activation development results of cultured oocytes, and as well as ROS and glutathione (GSH) levels in cumulus granulosa cells (CGs) were detected. Our results showed that the maturation rate of the SA 3D co-culture group was 85.41 %, that of the negative control (NC) group was 79.24 %, and that of the MGs co-culture group was 81.62 %. In SA 3D co-culture group, mitochondrial membrane potential level of oocytes was 1.6, ROS level was 19 and the ATP level was 1.7. While in NC group, mitochondrial membrane potential level of oocytes was 1.2, the ROS level was 52, and the ATP level was 0.4. The ROS level in the CGs of SA 3D co-culture group decreased by 1.5 times, and the glutathione content increased by 2.3 times. In the SA 3D co-culture group, GDF9 gene expression level was 2.0, and BMP15 gene expression level was 1.2. While in NC group, GDF9 gene expression level was 0.7, and BMP15 gene expression level was 0.6. The blastocyst rate in the SA 3D co-culture group was 41.4 %, and that in the NC group was 36.6 %. In conclusion, encapsulating MGs in SA gel and co-culturing them with porcine oocytes in 3D during IVM can improve the developmental potential of oocytes. This result will provide an important reference for improving the methods of in vitro maturation of oocytes.

{"title":"Follicle mural granulosa cells encapsulated in sodium alginate gel improve developmental competence of porcine oocytes of in vitro maturation","authors":"Qi Zhang, Yongfeng Zhou, Ran Ding, Qi Li, Xinglan An, Sheng Zhang, Ziyi Li","doi":"10.1016/j.theriogenology.2025.01.029","DOIUrl":"10.1016/j.theriogenology.2025.01.029","url":null,"abstract":"<div><div>The maturation of oocytes has an important impact on the subsequent development of the embryo. However, during the <em>in vitro</em> maturation (IVM) of oocytes, oocytes are separated from the follicular environment, resulting in a low maturation rate of oocytes <em>in vitro</em>. In order to improve maturation rate of IVM of porcine oocytes, this study was conducted to investigate using sodium alginate (SA) to encapsulate porcine mural granulosa cells (MGs) to develop an SA three-dimensional (3D) co-culture system for IVM of porcine oocytes. And, gene expression, reactive oxygen species (ROS), ATP level, mitochondrial membrane potential, parthenogenetic activation development results of cultured oocytes, and as well as ROS and glutathione (GSH) levels in cumulus granulosa cells (CGs) were detected. Our results showed that the maturation rate of the SA 3D co-culture group was 85.41 %, that of the negative control (NC) group was 79.24 %, and that of the MGs co-culture group was 81.62 %. In SA 3D co-culture group, mitochondrial membrane potential level of oocytes was 1.6, ROS level was 19 and the ATP level was 1.7. While in NC group, mitochondrial membrane potential level of oocytes was 1.2, the ROS level was 52, and the ATP level was 0.4. The ROS level in the CGs of SA 3D co-culture group decreased by 1.5 times, and the glutathione content increased by 2.3 times. In the SA 3D co-culture group, GDF9 gene expression level was 2.0, and BMP15 gene expression level was 1.2. While in NC group, GDF9 gene expression level was 0.7, and BMP15 gene expression level was 0.6. The blastocyst rate in the SA 3D co-culture group was 41.4 %, and that in the NC group was 36.6 %. In conclusion, encapsulating MGs in SA gel and co-culturing them with porcine oocytes in 3D during IVM can improve the developmental potential of oocytes. This result will provide an important reference for improving the methods of <em>in vitro</em> maturation of oocytes.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 52-59"},"PeriodicalIF":2.4,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143298629","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}

引用次数: 0

Expression of the glucose transporter 1 is associated with increased glucose uptake by granulosa cells during ovulation in mice

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-01-30 DOI: 10.1016/j.theriogenology.2025.01.028

Melissa Pansera, Neeraj Neeraj, Dayanada Siddappa, Yasmin Schuermann, Raj Duggavathi

The preovulatory luteinizing hormone surge is known to increase glucose uptake in ovulating follicles, but the underlying mechanisms have not been explored. Members of the Slc2a family of proteins called glucose transporters mediate glucose uptake in various cell types. Our objective was to characterize the expression pattern and temporal relationship with glucose uptake of the four best-characterized glucose transporters, Slc2a1-4 in mouse ovarian granulosa cells. Analyses of mRNA levels showed that Slc2a1 was induced in granulosa cells with a peak expression at 4h after human chorionic gonadotropin (hCG) treatment. We then examined signaling cascades involved in Slc2a1 expression by pharmacological inhibitors of the ERK1/2 and mTOR pathways. Inhibition of the ERK1/2 pathway by PD0325901 reduced Slc2a1 mRNA abundance demonstrating that the ERK1/2 signaling pathway is required for Slc2a1 expression. Conversely, inhibition of the mTOR pathway with rapamycin increased the Slc2a1 transcript level, which could be attributed to the compensatory hyperactivation of ERK1/2 activity. Bioinformatic analysis followed by chromatin immunoprecipitation showed that the transcription factor Cebpb binds to the Slc2a1 promoter in hCG-stimulated granulosa cells. Finally, the glucose uptake was higher in granulosa cells collected at 4h post-hCG than those collected at 0h hCG. These results indicate that the preovulatory LH surge increases glucose uptake in granulosa cells of the ovulating follicle by inducing Slc2a1 expression through the ERK1/2 pathway and its downstream effector transcription factor Cebpb.

{"title":"Expression of the glucose transporter 1 is associated with increased glucose uptake by granulosa cells during ovulation in mice","authors":"Melissa Pansera, Neeraj Neeraj, Dayanada Siddappa, Yasmin Schuermann, Raj Duggavathi","doi":"10.1016/j.theriogenology.2025.01.028","DOIUrl":"10.1016/j.theriogenology.2025.01.028","url":null,"abstract":"<div><div>The preovulatory luteinizing hormone surge is known to increase glucose uptake in ovulating follicles, but the underlying mechanisms have not been explored. Members of the Slc2a family of proteins called glucose transporters mediate glucose uptake in various cell types. Our objective was to characterize the expression pattern and temporal relationship with glucose uptake of the four best-characterized glucose transporters, Slc2a1-4 in mouse ovarian granulosa cells. Analyses of mRNA levels showed that <em>Slc2a1</em> was induced in granulosa cells with a peak expression at 4h after human chorionic gonadotropin (hCG) treatment. We then examined signaling cascades involved in <em>Slc2a1</em> expression by pharmacological inhibitors of the ERK1/2 and mTOR pathways. Inhibition of the ERK1/2 pathway by PD0325901 reduced <em>Slc2a1</em> mRNA abundance demonstrating that the ERK1/2 signaling pathway is required for <em>Slc2a1</em> expression. Conversely, inhibition of the mTOR pathway with rapamycin increased the <em>Slc2a1</em> transcript level, which could be attributed to the compensatory hyperactivation of ERK1/2 activity. Bioinformatic analysis followed by chromatin immunoprecipitation showed that the transcription factor Cebpb binds to the <em>Slc2a1</em> promoter in hCG-stimulated granulosa cells. Finally, the glucose uptake was higher in granulosa cells collected at 4h post-hCG than those collected at 0h hCG. These results indicate that the preovulatory LH surge increases glucose uptake in granulosa cells of the ovulating follicle by inducing <em>Slc2a1</em> expression through the ERK1/2 pathway and its downstream effector transcription factor Cebpb.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 13-20"},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143081117","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}

引用次数: 0

The regulatory repertoire of ZBTB16 in porcine immature spermatogonia

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-01-30 DOI: 10.1016/j.theriogenology.2025.01.026

Youjie Cui , Wei Liu , Xueni You , Wanying Li , Ruiqi Wu , Wenxian Zeng , Weijun Pang , Peng Wang , Yi Zheng

Spermatogenesis is a highly productive and intricate process occurring in testes that produces functional haploid sperm capable of fertilization therefore sustaining lifelong male fertility. A cornerstone of spermatogenesis is primitive spermatogonia, including spermatogonial stem cells (SSCs), that are able to self-renew and differentiate. The molecular mechanisms for spermatogonial self-renewal and differentiation in large domestic animals such as pigs, in comparison with their counterparts in mice, are poorly understood. In this study, we explored the expression pattern of ZBTB16 (a key transcription factor also known as PLZF) and its regulatory repertoire in porcine immature spermatogonia. We first co-stained ZBTB16 with spermatogonial/proliferative markers (DBA, SALL4, UCHL1 or Ki67) on testis sections from four ages of boars, demonstrating that ZBTB16⁺ cells in prepubertal porcine testes are a subpopulation of immature spermatogonia. Then, we knocked down ZBTB16 in enriched porcine immature spermatogonia, and the following RNA-sequencing (RNA-seq) analysis showed that ZBTB16 knockdown resulted in the manifest transcriptomic change, characterized by downregulation of genes related to spermatogonial self-renewal as well as upregulation of differentiation genes, corroborating ZBTB16 as a factor crucial to porcine spermatogonial self-renewal. Later, by performing a CUT&Tag analysis, we identified the genomic targets of ZBTB16 in porcine immature spermatogonia, and the final integrative analysis for RNA-seq and CUT&Tag data revealed the correlation of ZBTB16 with GDNF and mTOR signaling that facilitates porcine immature spermatogonial self-renewal. Altogether, our results enhance the understanding of molecular mechanisms for spermatogonial self-renewal in pigs, thereby facilitating the in vitro culture of porcine SSCs.

{"title":"The regulatory repertoire of ZBTB16 in porcine immature spermatogonia","authors":"Youjie Cui , Wei Liu , Xueni You , Wanying Li , Ruiqi Wu , Wenxian Zeng , Weijun Pang , Peng Wang , Yi Zheng","doi":"10.1016/j.theriogenology.2025.01.026","DOIUrl":"10.1016/j.theriogenology.2025.01.026","url":null,"abstract":"<div><div>Spermatogenesis is a highly productive and intricate process occurring in testes that produces functional haploid sperm capable of fertilization therefore sustaining lifelong male fertility. A cornerstone of spermatogenesis is primitive spermatogonia, including spermatogonial stem cells (SSCs), that are able to self-renew and differentiate. The molecular mechanisms for spermatogonial self-renewal and differentiation in large domestic animals such as pigs, in comparison with their counterparts in mice, are poorly understood. In this study, we explored the expression pattern of ZBTB16 (a key transcription factor also known as PLZF) and its regulatory repertoire in porcine immature spermatogonia. We first co-stained ZBTB16 with spermatogonial/proliferative markers (DBA, SALL4, UCHL1 or Ki67) on testis sections from four ages of boars, demonstrating that ZBTB16<sup>+</sup> cells in prepubertal porcine testes are a subpopulation of immature spermatogonia. Then, we knocked down <em>ZBTB16</em> in enriched porcine immature spermatogonia, and the following RNA-sequencing (RNA-seq) analysis showed that <em>ZBTB16</em> knockdown resulted in the manifest transcriptomic change, characterized by downregulation of genes related to spermatogonial self-renewal as well as upregulation of differentiation genes, corroborating ZBTB16 as a factor crucial to porcine spermatogonial self-renewal. Later, by performing a CUT&Tag analysis, we identified the genomic targets of ZBTB16 in porcine immature spermatogonia, and the final integrative analysis for RNA-seq and CUT&Tag data revealed the correlation of ZBTB16 with GDNF and mTOR signaling that facilitates porcine immature spermatogonial self-renewal. Altogether, our results enhance the understanding of molecular mechanisms for spermatogonial self-renewal in pigs, thereby facilitating the <em>in vitro</em> culture of porcine SSCs.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 21-32"},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143081121","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}

引用次数: 0

Fertility with cervical insemination and boar effects using hypothermic stored semen

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-01-30 DOI: 10.1016/j.theriogenology.2025.01.025

Florian Reckinger , Anne-Marie Luther , Anja Riesenbeck , Florian Sitzenstock , Ralf Wassmuth , Dagmar Waberski

Cold-storage of semen at 5 °C ± 1 °C provides a novel approach for counteracting antimicrobial resistance in artificial insemination (AI) of pigs. The first objective was to test the suitability of an AI boar population for cold semen storage. The second goal was to test cold semen storage in sow farms using the traditional cervical insemination (CAI) technique. For Experiment 1, semen from four repeated ejaculates of 20 AI boars was stored for 144h at 5 °C in AndroStar® Premium (APrem) and at 17 °C in Beltsville Thawing Solution (BTS). Sperm kinematics and acrosome integrity were assessed to identify inter- and intra-boar variability in sperm quality traits. All 80 semen samples met the minimum requirements for use in AI after long-term storage. Two boars demonstrated lower motility in 5 °C semen doses compared to 17 °C (p < 0.05). Intra-boar variation, analyzed as a coefficient of variance, was low in both semen storage groups. For Experiment 2, semen aliquots stored at 5 °C in APrem and at 17 °C in BTS were used for CAI of 579 sows in two farms under routine AI management conditions. Non-return rates, farrowing rates, and litter sizes were high in both farms and did not vary between the two semen storage groups (p > 0.05). In conclusion, the hypothermic semen storage concept is applicable with CAI and the use of relatively low sperm numbers. The in vitro data suggest a broad suitability of AI boars for semen storage at 5 °C.

{"title":"Fertility with cervical insemination and boar effects using hypothermic stored semen","authors":"Florian Reckinger , Anne-Marie Luther , Anja Riesenbeck , Florian Sitzenstock , Ralf Wassmuth , Dagmar Waberski","doi":"10.1016/j.theriogenology.2025.01.025","DOIUrl":"10.1016/j.theriogenology.2025.01.025","url":null,"abstract":"<div><div>Cold-storage of semen at 5 °C ± 1 °C provides a novel approach for counteracting antimicrobial resistance in artificial insemination (AI) of pigs. The first objective was to test the suitability of an AI boar population for cold semen storage. The second goal was to test cold semen storage in sow farms using the traditional cervical insemination (CAI) technique. For Experiment 1, semen from four repeated ejaculates of 20 AI boars was stored for 144h at 5 °C in AndroStar® Premium (APrem) and at 17 °C in Beltsville Thawing Solution (BTS). Sperm kinematics and acrosome integrity were assessed to identify inter- and intra-boar variability in sperm quality traits. All 80 semen samples met the minimum requirements for use in AI after long-term storage. Two boars demonstrated lower motility in 5 °C semen doses compared to 17 °C (p < 0.05). Intra-boar variation, analyzed as a coefficient of variance, was low in both semen storage groups. For Experiment 2, semen aliquots stored at 5 °C in APrem and at 17 °C in BTS were used for CAI of 579 sows in two farms under routine AI management conditions. Non-return rates, farrowing rates, and litter sizes were high in both farms and did not vary between the two semen storage groups (<em>p</em> > 0.05). In conclusion, the hypothermic semen storage concept is applicable with CAI and the use of relatively low sperm numbers. The <em>in vitro</em> data suggest a broad suitability of AI boars for semen storage at 5 °C.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 45-51"},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143168500","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}

引用次数: 0

Association between conditions affecting the vaginal portion of the cervix or the cranial portion of the vagina and reproductive performance in lactating dairy cows

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology

Pub Date : 2025-01-29 DOI: 10.1016/j.theriogenology.2025.01.024

M.B. Ugarte Marin , M.L. Newman , M.E. Hernandez , T.D. Gonzalez , C. Rouillon , E. Schmitt , T. Allard , N. Bliznyuk , K.N. Galvão , R.S. Bisinotto

Objectives

were to characterize the prevalence of anatomical abnormalities and pathological conditions involving the vaginal portion of the cervix or the cranial portion of the vagina and evaluate its association with reproductive outcomes and removal form herd in lactating dairy cows. Primiparous (n = 1675) and multiparous (n = 3594) cows from 3 herds were enrolled in a prospective cohort study. The vaginal portion of the cervix and the cranial portion of the vagina were examined using a digital vaginoscope in cows ranging from 0 to 70 days in milk (DIM) or at insemination. Proportion cows with well-defined longitudinal cervical folds (≥75 % of surface) was affected by the interaction between DIM category and parity, as it was smaller for primiparous compared with multiparous cows evaluated 0 to 35 DIM, but not from 36 to 70 DIM. Proportion of cows with well-defined longitudinal cervical folds increased with greater postpartum intervals, but no further change was observed after 36 to 50 DIM. Cows with calving problems were less likely to have well-defined longitudinal cervical folds from 36 to 50 DIM. Cows diagnosed with adhesions (0.5 %), double external uterine ostium (1.7 %), vaginal septum (0.6 %), masses (1.2 %), urovagina (0.7 %), and scar tissue (0.9 %) represented a small proportion of the population, however only cows with urovagina had greater odds of becoming reproductive ineligible. Proportion of cows with metritis was 5.1 % (0–7 DIM) and 7.4 % (8–21 DIM), which was not associated with outcomes of interest. Proportion of cows diagnosed with purulent vaginal discharge (PVD) was 11.9 % (22–35 DIM), 8.5 % (36–50 DIM), and 6.3 % (51–70 DIM). Cows with PVD had reduced risk of receiving AI (51–70 DIM), smaller proportion of cows pregnant by 305 DIM (36–70 DIM), and increased odds of reproductive culling (36–70 DIM). The presence of bloody discharge at the day of AI was negatively associated with the odds of becoming pregnant.

{"title":"Association between conditions affecting the vaginal portion of the cervix or the cranial portion of the vagina and reproductive performance in lactating dairy cows","authors":"M.B. Ugarte Marin , M.L. Newman , M.E. Hernandez , T.D. Gonzalez , C. Rouillon , E. Schmitt , T. Allard , N. Bliznyuk , K.N. Galvão , R.S. Bisinotto","doi":"10.1016/j.theriogenology.2025.01.024","DOIUrl":"10.1016/j.theriogenology.2025.01.024","url":null,"abstract":"<div><h3>Objectives</h3><div>were to characterize the prevalence of anatomical abnormalities and pathological conditions involving the vaginal portion of the cervix or the cranial portion of the vagina and evaluate its association with reproductive outcomes and removal form herd in lactating dairy cows. Primiparous (n = 1675) and multiparous (n = 3594) cows from 3 herds were enrolled in a prospective cohort study. The vaginal portion of the cervix and the cranial portion of the vagina were examined using a digital vaginoscope in cows ranging from 0 to 70 days in milk (DIM) or at insemination. Proportion cows with well-defined longitudinal cervical folds (≥75 % of surface) was affected by the interaction between DIM category and parity, as it was smaller for primiparous compared with multiparous cows evaluated 0 to 35 DIM, but not from 36 to 70 DIM. Proportion of cows with well-defined longitudinal cervical folds increased with greater postpartum intervals, but no further change was observed after 36 to 50 DIM. Cows with calving problems were less likely to have well-defined longitudinal cervical folds from 36 to 50 DIM. Cows diagnosed with adhesions (0.5 %), double external uterine ostium (1.7 %), vaginal septum (0.6 %), masses (1.2 %), urovagina (0.7 %), and scar tissue (0.9 %) represented a small proportion of the population, however only cows with urovagina had greater odds of becoming reproductive ineligible. Proportion of cows with metritis was 5.1 % (0–7 DIM) and 7.4 % (8–21 DIM), which was not associated with outcomes of interest. Proportion of cows diagnosed with purulent vaginal discharge (PVD) was 11.9 % (22–35 DIM), 8.5 % (36–50 DIM), and 6.3 % (51–70 DIM). Cows with PVD had reduced risk of receiving AI (51–70 DIM), smaller proportion of cows pregnant by 305 DIM (36–70 DIM), and increased odds of reproductive culling (36–70 DIM). The presence of bloody discharge at the day of AI was negatively associated with the odds of becoming pregnant.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"236 ","pages":"Pages 82-95"},"PeriodicalIF":2.4,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143349364","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}

引用次数: 0