Pub Date : 2024-03-13DOI: 10.1016/j.anireprosci.2024.107455
Pablo Torres, Daniela Marina Malcervelli, Jonathan Hernán Cartelle, María Laura Fischman, Humberto Osvaldo Cisale
Boar sperm is highly susceptible to cold damage. When temperature drops to 5°C, the plasmatic membrane is destabilized. The freezing process causes a reduction of the fertility window because frozen/thawed boar sperm has less survivability. The aim of this work was to analyze the effect on sperm characteristics and response to capacitation stimuli of cooling to 5°C using a controlled protocol. Also, we evaluated if the addition of Glycerol 2% or 3% at 5°C was able to modify these parameters. For this purpose, we assessed motility, plasmatic membrane integrity and acrosomal membrane status. Capacitation was induced using Tyrode´s capacitating medium (TCM) and assessed by chlortetracycline stain and induction of acrosomal reaction with Progesterone. Motility patterns were analyzed using a CASA system. These tests were performed at three different points of the freezing curve: 37°C; 17°C and 5°C. Response to TCM vs TBM was only significant at 37°C. While at 37°C and 17°C capacitated sperm was below 20%, at 5°C reached 50% both in the TBM and TCM. CASA analysis showed that spermatozoa exposed to TCM had higher LIN and WOB than those in TBM. All parameters were similar in the Glycerol concentrations studied. These results suggest that the chilling process may be causing an effect similar to cryocapacitation along the cooling curve, starting subtle at 17°C and reaching 50% of the sperm population at 5°C, being independent of Glycerol concentration.
{"title":"Cold-induced damage in boar sperm cooled to 5°C in Lactose Egg Yolk (LEY) medium with different Glycerol concentrations","authors":"Pablo Torres, Daniela Marina Malcervelli, Jonathan Hernán Cartelle, María Laura Fischman, Humberto Osvaldo Cisale","doi":"10.1016/j.anireprosci.2024.107455","DOIUrl":"10.1016/j.anireprosci.2024.107455","url":null,"abstract":"<div><p>Boar sperm is highly susceptible to cold damage. When temperature drops to 5°C, the plasmatic membrane is destabilized. The freezing process causes a reduction of the fertility window because frozen/thawed boar sperm has less survivability. The aim of this work was to analyze the effect on sperm characteristics and response to capacitation stimuli of cooling to 5°C using a controlled protocol. Also, we evaluated if the addition of Glycerol 2% or 3% at 5°C was able to modify these parameters. For this purpose, we assessed motility, plasmatic membrane integrity and acrosomal membrane status. Capacitation was induced using Tyrode´s capacitating medium (TCM) and assessed by chlortetracycline stain and induction of acrosomal reaction with Progesterone. Motility patterns were analyzed using a CASA system. These tests were performed at three different points of the freezing curve: 37°C; 17°C and 5°C. Response to TCM vs TBM was only significant at 37°C. While at 37°C and 17°C capacitated sperm was below 20%, at 5°C reached 50% both in the TBM and TCM. CASA analysis showed that spermatozoa exposed to TCM had higher LIN and WOB than those in TBM. All parameters were similar in the Glycerol concentrations studied. These results suggest that the chilling process may be causing an effect similar to cryocapacitation along the cooling curve, starting subtle at 17°C and reaching 50% of the sperm population at 5°C, being independent of Glycerol concentration.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140269894","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}
Pub Date : 2024-03-13DOI: 10.1016/j.anireprosci.2024.107457
Hui Xu , Yu Cai , Hua Yang , Shanglai Li , Peiyong Chen , Zongyou Wei , Feng Wang , Ziyu Wang , Yanli Zhang
The anterior pituitary plays a critical role in the endocrine system, contains gonadotrophs, which regulate reproductive efficiency by secreting follicle-stimulating hormone (FSH) and luteinizing hormone (LH). PPP2R2A is a serine-threonine phosphatase that regulates reproductive functions in both females and males, its function in pituitary cells remain unclear. Hu sheep is a highly prolific breed, which makes it suitable for studying reproductive mechanisms. In this study, the relative abundances of PPP2R2A mRNA expression were higher in the pituitary of high-prolificacy (HF) Hu sheep compared to those of low-prolificacy (LF) Hu sheep. Additionally, we demonstrated that PPP2R2A promotes pituitary cell proliferation and gonadotropin secretion using the EdU assay and ELISA, respectively. Moreover, it inhibits pituitary cell apoptosis using flow cytometry. Furthermore, PPP2R2A may affect pituitary cell function by regulating the AKT/mTOR signaling pathway. In summary, our findings suggest that PPP2R2A may play a role in regulating pituitary function and influencing the secretion of gonadotropins.
{"title":"PPP2R2A promotes Hu sheep pituitary cell proliferation and gonadotropin secretion associated with prolificacy","authors":"Hui Xu , Yu Cai , Hua Yang , Shanglai Li , Peiyong Chen , Zongyou Wei , Feng Wang , Ziyu Wang , Yanli Zhang","doi":"10.1016/j.anireprosci.2024.107457","DOIUrl":"10.1016/j.anireprosci.2024.107457","url":null,"abstract":"<div><p>The anterior pituitary plays a critical role in the endocrine system, contains gonadotrophs, which regulate reproductive efficiency by secreting follicle-stimulating hormone (FSH) and luteinizing hormone (LH). PPP2R2A is a serine-threonine phosphatase that regulates reproductive functions in both females and males, its function in pituitary cells remain unclear. Hu sheep is a highly prolific breed, which makes it suitable for studying reproductive mechanisms. In this study, the relative abundances of PPP2R2A mRNA expression were higher in the pituitary of high-prolificacy (HF) Hu sheep compared to those of low-prolificacy (LF) Hu sheep. Additionally, we demonstrated that PPP2R2A promotes pituitary cell proliferation and gonadotropin secretion using the EdU assay and ELISA, respectively. Moreover, it inhibits pituitary cell apoptosis using flow cytometry. Furthermore, PPP2R2A may affect pituitary cell function by regulating the AKT/mTOR signaling pathway. In summary, our findings suggest that PPP2R2A may play a role in regulating pituitary function and influencing the secretion of gonadotropins.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140280513","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}
Pub Date : 2024-03-12DOI: 10.1016/j.anireprosci.2024.107452
Rodney D. Geisert , Fuller W. Bazer , Caroline G. Lucas , Caroline A. Pfeiffer , Ashley E. Meyer , Riley Sullivan , Destiny N. Johns , Mariana Sponchiado , Randall S. Prather
Maternal recognition of pregnancy (MRP) is a term utilized in mammals to describe pathways in which the conceptus alters the endometrial environment to prevent regression of corpora lutea to ensure continued production of progesterone (P4) required for establishment and maintenance of pregnancy. For nearly 40 years after publication of the endocrine/exocrine theory, conceptus estrogen (E2) was considered the primary maternal recognition signal in the pig. Conceptus production of prostaglandin E2 (PGE2) was also considered to be a major factor in preventing luteolysis. An addition to E2 and PGE2, pig conceptuses produce interleukin 1B2 (IL1B2) and interferons (IFN) delta (IFND) and gamma (IFNG). The present review provides brief history of the discovery of E2, PGs and IFNS which led to research investigating the role of these conceptus secreted factors in establishing and maintaining pregnancy in the pig. The recent utilization of gene editing technology allowed a more direct approach to investigate the in vivo roles of IL1B2, E2, PGE2, AND IFNG for establishment of pregnancy. These studies revealed unknown functions for IFNG and ILB2 in addition to PGE2 and E2. Thus, pregnancy recognition signal is via a servomechanism in requiring sequential effects of P4, E2, IL1B2, PGE2 and IFNG. Results indicate that the original established dogma for the role of conceptus E2 and PGs in MRP is a far too simplified model that involves the interplay of numerous mechanisms for inhibiting luteolysis, inducing critical elongation of the conceptuses and resolution of inflammation in pigs.
{"title":"Maternal recognition of pregnancy in the pig: A servomechanism involving sex steroids, cytokines and prostaglandins","authors":"Rodney D. Geisert , Fuller W. Bazer , Caroline G. Lucas , Caroline A. Pfeiffer , Ashley E. Meyer , Riley Sullivan , Destiny N. Johns , Mariana Sponchiado , Randall S. Prather","doi":"10.1016/j.anireprosci.2024.107452","DOIUrl":"10.1016/j.anireprosci.2024.107452","url":null,"abstract":"<div><p>Maternal recognition of pregnancy (MRP) is a term utilized in mammals to describe pathways in which the conceptus alters the endometrial environment to prevent regression of corpora lutea to ensure continued production of progesterone (P4) required for establishment and maintenance of pregnancy. For nearly 40 years after publication of the endocrine/exocrine theory, conceptus estrogen (E2) was considered the primary maternal recognition signal in the pig. Conceptus production of prostaglandin E2 (PGE2) was also considered to be a major factor in preventing luteolysis. An addition to E2 and PGE2, pig conceptuses produce interleukin 1B2 (IL1B2) and interferons (IFN) delta (IFND) and gamma (IFNG). The present review provides brief history of the discovery of E2, PGs and IFNS which led to research investigating the role of these conceptus secreted factors in establishing and maintaining pregnancy in the pig. The recent utilization of gene editing technology allowed a more direct approach to investigate the <em>in vivo</em> roles of IL1B2, E2, PGE2, AND IFNG for establishment of pregnancy. These studies revealed unknown functions for IFNG and ILB2 in addition to PGE2 and E2. Thus, pregnancy recognition signal is via a servomechanism in requiring sequential effects of P4, E2, IL1B2, PGE2 and IFNG. Results indicate that the original established dogma for the role of conceptus E2 and PGs in MRP is a far too simplified model that involves the interplay of numerous mechanisms for inhibiting luteolysis, inducing critical elongation of the conceptuses and resolution of inflammation in pigs.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140128993","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}
Pub Date : 2024-03-12DOI: 10.1016/j.anireprosci.2024.107454
Erfan Akbari Nargesi , Danial Gorouhi
This study aimed to compare the effectiveness of the ovarian lavage / artificial insemination method with the traditional hormonal administration and fertilization methods over the artificial reproduction of the northern pike (Esox lucius). For this purpose, groups of five females were treated as follows: intraperitoneal injection of saline (C1); ovarian lavage with saline (C2); intraperitoneal injection of carp pituitary extract (CPE, T1); ovarian lavage with CPE (T2); intraperitoneal injection of CPE and ovarian lavage with semen after 72 h (T3); ovarian lavage with CPE and ovarian lavage with semen after 72 h (T4). According to the results, no fish ovulated in the control groups (C1 and C2). There were no significant differences (n.s.) among experimental treatments (P > 0.05, n.s.) in the reproductive parameters, such as latency time, ovulation rate, stripped egg amount, and pseudo-gonadosomatic index. The lowest fertilization rate (54.8%) was observed in the T4 treatment and significantly differed from the T1 and T2 treatments (P < 0.05). Moreover, the highest survival at swim-up stage was measured in the T4 treatment with a significant difference compared to the T1 group (P < 0.05). The survival at the eyed-egg stage, hatching rate, and malformations were similar (P > 0.05, n.s.) in all applied strategies. The results demonstrated that the ovarian lavage / artificial insemination method could be applied to control northern pike reproduction, like the traditional fertilization method. Consequently, this novel technique can be suggested as an alternative strategy to facilitate the hatchery operations in the controlled reproduction of this species.
{"title":"Ovarian lavage / artificial insemination method as an alternative approach for controlled reproduction of the northern pike (Esox lucius)","authors":"Erfan Akbari Nargesi , Danial Gorouhi","doi":"10.1016/j.anireprosci.2024.107454","DOIUrl":"10.1016/j.anireprosci.2024.107454","url":null,"abstract":"<div><p>This study aimed to compare the effectiveness of the ovarian lavage / artificial insemination method with the traditional hormonal administration and fertilization methods over the artificial reproduction of the northern pike (<em>Esox lucius</em>). For this purpose, groups of five females were treated as follows: intraperitoneal injection of saline (C1); ovarian lavage with saline (C2); intraperitoneal injection of carp pituitary extract (CPE, T1); ovarian lavage with CPE (T2); intraperitoneal injection of CPE and ovarian lavage with semen after 72 h (T3); ovarian lavage with CPE and ovarian lavage with semen after 72 h (T4). According to the results, no fish ovulated in the control groups (C1 and C2). There were no significant differences (n.s.) among experimental treatments (<em>P</em> > 0.05, n.s.) in the reproductive parameters, such as latency time, ovulation rate, stripped egg amount, and pseudo-gonadosomatic index. The lowest fertilization rate (54.8%) was observed in the T4 treatment and significantly differed from the T1 and T2 treatments (<em>P</em> < 0.05). Moreover, the highest survival at swim-up stage was measured in the T4 treatment with a significant difference compared to the T1 group (<em>P</em> < 0.05). The survival at the eyed-egg stage, hatching rate, and malformations were similar (<em>P</em> > 0.05, n.s.) in all applied strategies. The results demonstrated that the ovarian lavage / artificial insemination method could be applied to control northern pike reproduction, like the traditional fertilization method. Consequently, this novel technique can be suggested as an alternative strategy to facilitate the hatchery operations in the controlled reproduction of this species.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140129909","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}
Pub Date : 2024-03-08DOI: 10.1016/j.anireprosci.2024.107451
Deepak Shinde, Shilpa K. Bhat, C.B. Ganesh
In vertebrates, opioid peptides are thought to be involved in the regulation of reproduction; however, the significance of enkephalins in testicular function remains unclear. We examined the influence of δ-opioid receptor agonist leucine enkephalin (L-ENK) on the hypophysial-testicular axis of the cichlid fish Oreochromis mossambicus. Treatment with a low dose of L-ENK (60 µg) caused a significant increase in the numbers of primary and secondary spermatocytes and early and late spermatids, concomitant with intense immunolabelling of testicular androgen receptors, but did not significantly alter serum luteinizing hormone (LH) and 11-ketotestosterone (11-KT) levels compared to those of controls. Nevertheless, treatment with a high dose of L-ENK (200 µg) caused a significant reduction in the numbers of secondary spermatocytes as well as late spermatids associated with marginal immunolabelling of androgen receptors and significantly lower concentrations of serum 11-KT and LH compared to controls. In addition, the serum cortisol level was not affected in low-dose L-ENK-treated fish, but its level was significantly increased in the high-dose L-ENK-treated group. Together, these findings indicate that a low dose of L-ENK stimulates the germ cells at the meiosis stage and promotes further stages of spermatogenesis, whereas a high concentration of L-ENK inhibits spermatogenesis at the advanced stages. This effect appears to be mediated through the suppression of testicular steroidogenesis and the reduction of LH release in the pituitary gland of tilapia. The findings also suggest that elevated L-ENK levels in teleosts may exert their inhibitory influence on the hypophysial-testicular axis via glucocorticoids.
{"title":"The opioid peptide leucine enkephalin modulates hypothalamic-hypophysial axis in the cichlid fish Oreochromis mossambicus","authors":"Deepak Shinde, Shilpa K. Bhat, C.B. Ganesh","doi":"10.1016/j.anireprosci.2024.107451","DOIUrl":"https://doi.org/10.1016/j.anireprosci.2024.107451","url":null,"abstract":"<div><p>In vertebrates, opioid peptides are thought to be involved in the regulation of reproduction; however, the significance of enkephalins in testicular function remains unclear. We examined the influence of δ-opioid receptor agonist leucine enkephalin (L-ENK) on the hypophysial-testicular axis of the cichlid fish <em>Oreochromis mossambicus</em>. Treatment with a low dose of L-ENK (60 µg) caused a significant increase in the numbers of primary and secondary spermatocytes and early and late spermatids, concomitant with intense immunolabelling of testicular androgen receptors, but did not significantly alter serum luteinizing hormone (LH) and 11-ketotestosterone (11-KT) levels compared to those of controls. Nevertheless, treatment with a high dose of L-ENK (200 µg) caused a significant reduction in the numbers of secondary spermatocytes as well as late spermatids associated with marginal immunolabelling of androgen receptors and significantly lower concentrations of serum 11-KT and LH compared to controls. In addition, the serum cortisol level was not affected in low-dose L-ENK-treated fish, but its level was significantly increased in the high-dose L-ENK-treated group. Together, these findings indicate that a low dose of L-ENK stimulates the germ cells at the meiosis stage and promotes further stages of spermatogenesis, whereas a high concentration of L-ENK inhibits spermatogenesis at the advanced stages. This effect appears to be mediated through the suppression of testicular steroidogenesis and the reduction of LH release in the pituitary gland of tilapia. The findings also suggest that elevated L-ENK levels in teleosts may exert their inhibitory influence on the hypophysial-testicular axis via glucocorticoids.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140122437","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}
Pub Date : 2024-03-02DOI: 10.1016/j.anireprosci.2024.107450
Stanimir Yotov , Desislava Abadjieva , Anatoli Atanasov , Boyana Ivanova , Paulina Taushanova , Ivan Fasulkov , Elena Kistanova
The quality of the separated fractions in sex-sorted semen is very important for the success of the artificial insemination. This study aimed to evaluate some in vitro characteristics (DNA quantity, kinematic parameters and enzymes activity) of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and toll-like receptors (TLR)7/8 ligand R848. The ejaculates from six rams were collected by artificial vagina and subjected to a computer-assisted semen analysis (CASA). Total motility and percentage of the sperms with rapid and medium progressivity or non-progressivity in whole ejaculates and in X and Y fractions were analyzed. Activity of the enzymes ALP, GGT, CK, LDH and accumulation of lactate in the seminal plasma of ejaculates and in the environmental fluid of sexed spermatozoa were measured by biochemical analyzer. DNA was isolated from precipitated spermatozoa, and its quantity was measured. For both protocols the DNA mass from X-bearing fractions was higher, than from Y-bearing fractions. The high total motility of X- and Y-bearing spermatozoa as well as greater percent sperms with progressive motility were observed after use of BSA protocol. The application of TLR7/8 ligand R848 protocol led to reducing of Y-sperm motility and enhancement of non-progressivity in both fractions, which corresponded to the determined high amount of the extracellular lactate. For both methods, the significantly reduced activity of enzymes in the X and Y spermatozoa environmental fluids was established. Both protocols produce X- and Y-sperm fractions with satisfactory quality (over 80% total motility and over 50% rapid and medium progressive spermatozoa in each fraction).
性别分选精液中分离部分的质量对人工授精的成功非常重要。本研究旨在评估用牛血清白蛋白(BSA)柱和收费样受体(TLR)7/8配体 R848 分选的 X 型和 Y 型公羊精子的一些体外特征(DNA 数量、运动参数和酶活性)。通过人工阴道采集了六只公羊的射精,并对其进行了计算机辅助精液分析(CASA)。分析了整个精液以及 X 和 Y 部分精子的总运动能力、快速和中等进展性或非进展性精子的百分比。生化分析仪测量了射精的精浆和有性别精子环境液中 ALP、GGT、CK、LDH 等酶的活性和乳酸的积累。从沉淀的精子中分离出 DNA 并测量其数量。在两种方案中,X 型精子的 DNA 质量都高于 Y 型精子。使用 BSA 方案后,X 型和 Y 型精子的总运动能力较高,具有渐进运动能力的精子百分比也较高。使用 TLR7/8 配体 R848 方案会降低 Y 型精子的运动能力,并提高两个馏分中的非运动能力,这与确定的高细胞外乳酸盐含量相符。对于这两种方法,X 精子和 Y 精子环境液中酶的活性都明显降低。两种方法都能产生质量令人满意的X和Y精子馏分(每个馏分中总运动率超过80%,快速和中等进展精子超过50%)。
{"title":"In vitro characteristics of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and TLR7/8 ligand R848","authors":"Stanimir Yotov , Desislava Abadjieva , Anatoli Atanasov , Boyana Ivanova , Paulina Taushanova , Ivan Fasulkov , Elena Kistanova","doi":"10.1016/j.anireprosci.2024.107450","DOIUrl":"10.1016/j.anireprosci.2024.107450","url":null,"abstract":"<div><p>The quality of the separated fractions in sex-sorted semen is very important for the success of the artificial insemination. This study aimed to evaluate some in vitro characteristics (DNA quantity, kinematic parameters and enzymes activity) of X- and Y-bearing ram spermatozoa sorted by bovine serum albumin (BSA) column and toll-like receptors (TLR)7/8 ligand R848. The ejaculates from six rams were collected by artificial vagina and subjected to a computer-assisted semen analysis (CASA). Total motility and percentage of the sperms with rapid and medium progressivity or non-progressivity in whole ejaculates and in X and Y fractions were analyzed. Activity of the enzymes ALP, GGT, CK, LDH and accumulation of lactate in the seminal plasma of ejaculates and in the environmental fluid of sexed spermatozoa were measured by biochemical analyzer. DNA was isolated from precipitated spermatozoa, and its quantity was measured. For both protocols the DNA mass from X-bearing fractions was higher, than from Y-bearing fractions. The high total motility of X- and Y-bearing spermatozoa as well as greater percent sperms with progressive motility were observed after use of BSA protocol. The application of TLR7/8 ligand R848 protocol led to reducing of Y-sperm motility and enhancement of non-progressivity in both fractions, which corresponded to the determined high amount of the extracellular lactate. For both methods, the significantly reduced activity of enzymes in the X and Y spermatozoa environmental fluids was established. Both protocols produce X- and Y-sperm fractions with satisfactory quality (over 80% total motility and over 50% rapid and medium progressive spermatozoa in each fraction).</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-03-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140054892","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}
Early embryonic mortality resulting from insufficient interaction between the embryo and the uterus leads to the failure of pregnancy in livestock animals. Thus, it is imperative to comprehend the multifaceted process of implantation at molecular levels, which requires synchronized feto-maternal interaction. The in-vitro models serve as valuable tools to investigate the specific stages of implantation. The present study was undertaken to develop a simple method to isolate and culture the primary buffalo endometrial epithelial cells (pBuEECs), followed by proteome profiling of the proliferating cells. Collagenase I was used to separate uterine epithelial cells (UECs) from the ipsilateral uterine horn, and then the cells were separated using a cell strainer. After being seeded on culture plates, UECs developed colonies with characteristic epithelial shape and expressed important markers such as cytokeratin 18 (KRT18), progesterone receptor (PGR), β-estrogen receptor (ESR1), and leukemia inhibitory factor (LIF), which were confirmed by PCR. The purity of epithelial cells was assessed using cytokeratin 18 immunostaining, which indicated approximately 99% purity in cultured cells. The proteome profiling of pBuEECs via high-throughput tandem mass spectrometry (MS), identified a total of 3383 proteins. Bioinformatics analysis revealed enrichment in various biological processes, including cellular processes, metabolic processes, biological regulation, localization, signaling, and developmental processes. Moreover, the KEGG pathway analysis highlighted associations with the ribosome, proteosome, oxidative phosphorylation, spliceosome, and cytoskeleton regulation pathways. In conclusion, these well characterized cells offer valuable in-vitro model to enhance the understanding of implantation and uterine pathophysiology in livestock animals, particularly buffaloes.
{"title":"Simple method for isolation and culture of primary buffalo (Bubalus bubalis) endometrial epithelial cells (pBuEECs) and its characterization using high throughput proteomics approach","authors":"Shradha Jamwal , Nikunj Tyagi , Jaideep Kumar , Jai Kumar Kaushik , Sudarshan Kumar , Ashok Kumar Mohanty","doi":"10.1016/j.anireprosci.2024.107449","DOIUrl":"10.1016/j.anireprosci.2024.107449","url":null,"abstract":"<div><p>Early embryonic mortality resulting from insufficient interaction between the embryo and the uterus leads to the failure of pregnancy in livestock animals. Thus, it is imperative to comprehend the multifaceted process of implantation at molecular levels, which requires synchronized feto-maternal interaction. The <em>in-vitro</em> models serve as valuable tools to investigate the specific stages of implantation. The present study was undertaken to develop a simple method to isolate and culture the primary buffalo endometrial epithelial cells (pBuEECs), followed by proteome profiling of the proliferating cells. Collagenase I was used to separate uterine epithelial cells (UECs) from the ipsilateral uterine horn, and then the cells were separated using a cell strainer. After being seeded on culture plates, UECs developed colonies with characteristic epithelial shape and expressed important markers such as cytokeratin 18 (KRT18), progesterone receptor (PGR), β-estrogen receptor (ESR1), and leukemia inhibitory factor (LIF), which were confirmed by PCR. The purity of epithelial cells was assessed using cytokeratin 18 immunostaining, which indicated approximately 99% purity in cultured cells. The proteome profiling of pBuEECs via high-throughput tandem mass spectrometry (MS), identified a total of 3383 proteins. Bioinformatics analysis revealed enrichment in various biological processes, including cellular processes, metabolic processes, biological regulation, localization, signaling, and developmental processes. Moreover, the KEGG pathway analysis highlighted associations with the ribosome, proteosome, oxidative phosphorylation, spliceosome, and cytoskeleton regulation pathways. In conclusion, these well characterized cells offer valuable <em>in-vitro</em> model to enhance the understanding of implantation and uterine pathophysiology in livestock animals, particularly buffaloes.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140055011","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}
Pub Date : 2024-02-27DOI: 10.1016/j.anireprosci.2024.107448
Jingle Jiang, Di Hu, Enle Pei
The egg production of captive African penguins differs considerably between individuals. An understanding of the physiological differences in African penguins with relatively greater and lesser egg production is meaningful for the captive breeding program of this endangered species. The objective of this study was to investigate differential microbial composition and metabolites in captive African penguins with different egg production. Fecal samples were collected from captive female African penguins during the breeding season. The results of 16 S rRNA gene sequencing showed that African penguins with different egg production had similar microbial diversities, whereas a significant difference was observed between their microbial community structure. African penguins with relatively greater egg production exhibited a higher relative abundance of Alphaproteobacteria, Rhizobiales, Bradyrhizobiaceae, Bradyrhizobium and Bosea. Meanwhile, penguins with relatively lesser egg production had an increased proportion of Klebsiella and Plesiomonas. We further identified a total of 1858 metabolites in female African penguins by liquid chromatography-mass spectrometry analysis. Among these metabolites, 13 kinds of metabolites were found to be significantly differential between African penguins with different egg production. In addition, the correlation analysis revealed that the egg production had significant correlations with most of the differential microbial bacteria and metabolites. Our findings might aid in understanding the potential mechanism underlying the phenomenon of abnormal egg production in captive African penguins, and provide novel insights into the relationship between gut microbiota and reproduction in penguins.
人工饲养的非洲企鹅的产卵量在个体之间存在很大差异。了解产蛋量相对较高和较低的非洲企鹅的生理差异,对这一濒危物种的人工繁殖计划很有意义。本研究的目的是调查不同产蛋量的圈养非洲企鹅体内不同的微生物组成和代谢物。研究人员在繁殖季节收集了圈养雌性非洲企鹅的粪便样本。16 S rRNA基因测序结果表明,产卵量不同的非洲企鹅具有相似的微生物多样性,但其微生物群落结构存在显著差异。产蛋量相对较高的非洲企鹅表现出较高的Alphaproteobacteria、Rhizobiales、Bradyrhizobiaceae、Bradyrhizobium和Bosea的相对丰度。与此同时,产卵量相对较少的企鹅体内克雷伯氏菌和普莱西单胞菌的比例有所增加。通过液相色谱-质谱分析,我们进一步鉴定了雌性非洲企鹅体内的1858种代谢物。在这些代谢物中,有13种代谢物在不同产蛋量的非洲企鹅之间存在显著差异。此外,相关分析表明,产蛋量与大多数不同的微生物细菌和代谢物有明显的相关性。我们的研究结果可能有助于理解圈养非洲企鹅异常产蛋现象的潜在机制,并为企鹅肠道微生物群与繁殖之间的关系提供了新的见解。
{"title":"Integrated omics analysis reveals a correlation between gut microbiota and egg production in captive African penguins (Spheniscus demersus)","authors":"Jingle Jiang, Di Hu, Enle Pei","doi":"10.1016/j.anireprosci.2024.107448","DOIUrl":"https://doi.org/10.1016/j.anireprosci.2024.107448","url":null,"abstract":"<div><p>The egg production of captive African penguins differs considerably between individuals. An understanding of the physiological differences in African penguins with relatively greater and lesser egg production is meaningful for the captive breeding program of this endangered species. The objective of this study was to investigate differential microbial composition and metabolites in captive African penguins with different egg production. Fecal samples were collected from captive female African penguins during the breeding season. The results of 16 S rRNA gene sequencing showed that African penguins with different egg production had similar microbial diversities, whereas a significant difference was observed between their microbial community structure. African penguins with relatively greater egg production exhibited a higher relative abundance of Alphaproteobacteria, Rhizobiales, Bradyrhizobiaceae, <em>Bradyrhizobium</em> and <em>Bosea</em>. Meanwhile, penguins with relatively lesser egg production had an increased proportion of <em>Klebsiella</em> and <em>Plesiomonas</em>. We further identified a total of 1858 metabolites in female African penguins by liquid chromatography-mass spectrometry analysis. Among these metabolites, 13 kinds of metabolites were found to be significantly differential between African penguins with different egg production. In addition, the correlation analysis revealed that the egg production had significant correlations with most of the differential microbial bacteria and metabolites. Our findings might aid in understanding the potential mechanism underlying the phenomenon of abnormal egg production in captive African penguins, and provide novel insights into the relationship between gut microbiota and reproduction in penguins.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139999381","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}
Pub Date : 2024-02-24DOI: 10.1016/j.anireprosci.2024.107439
Aderson M. Viana Neto , Denise D. Guerreiro , Jorge A.M. Martins , F.ábio R. Vasconcelos , R.évila B.F. Melo , Ana Luiza M.C. S. Velho , Marta Neila-Montero , Rafael Montes-Garrido , Celso S. Nagano , Airton A. Araújo , Arlindo A. Moura
The present study evaluated the effects of heat stress on reproductive parameters of hairy rams. Six animals were subjected to scrotal insulation during four consecutive nights (6 PM – 6 AM). Day (D) 0 was the first day of insulation. Scrotal circumference increased from 30.5 ± 0.3 cm (at pre-insulation) to 31.8 ± 0.4 cm on D4, decreased 3.9 cm on D28, returning to 30.6 ± 0.6 cm on D57. Sperm concentration decreased from 3.7 ± 0.12 ×109 sperm/mL before insulation to 2.6 ± 0.1 ×109 on D23, returning to normal on D57. Sperm motility averaged 75 ± 2.9% before insulation, was undetectable on D23, and became normal on D77. Sperm with normal morphology reached 5.9 ± 2.6% on D35 but recovered (86.8 ± 2.1%) on D91. Sperm DNA integrity decreased from 86.5 ± 4.7% before insulation to 11.1 ± 3.7% on D63, returning to pre-insulation values on D120. Sperm BSP immunostaining was reduced after scrotal insulation. Variations in seminal protein abundances coincided with changes in sperm parameters. Seminal plasma superoxide dismutase, carboxypeptidase Q-precursor and NPC intracellular cholesterol transporter 2 decreased on D18, returning to normal after D28. Albumin, inhibitor of carbonic anhydrase precursor, EGF-like repeat and discoid I-like domain-containing protein 3 and polymeric immunoglobulin receptor increased after insulation. In summary, intermittent scrotal insulation drastically altered ram sperm attributes and seminal proteins, especially those associated with oxidative stress. Knowledge of animal´s response to thermal stress is vital in the scenario of climate changes.
{"title":"Sperm traits and seminal plasma proteome of locally adapted hairy rams subjected to intermittent scrotal insulation","authors":"Aderson M. Viana Neto , Denise D. Guerreiro , Jorge A.M. Martins , F.ábio R. Vasconcelos , R.évila B.F. Melo , Ana Luiza M.C. S. Velho , Marta Neila-Montero , Rafael Montes-Garrido , Celso S. Nagano , Airton A. Araújo , Arlindo A. Moura","doi":"10.1016/j.anireprosci.2024.107439","DOIUrl":"10.1016/j.anireprosci.2024.107439","url":null,"abstract":"<div><p>The present study evaluated the effects of heat stress on reproductive parameters of hairy rams. Six animals were subjected to scrotal insulation during four consecutive nights (6 PM – 6 AM). Day (D) 0 was the first day of insulation. Scrotal circumference increased from 30.5 ± 0.3 cm (at pre-insulation) to 31.8 ± 0.4 cm on D4, decreased 3.9 cm on D28, returning to 30.6 ± 0.6 cm on D57. Sperm concentration decreased from 3.7 ± 0.12 ×10<sup>9</sup> sperm/mL before insulation to 2.6 ± 0.1 ×10<sup>9</sup> on D23, returning to normal on D57. Sperm motility averaged 75 ± 2.9% before insulation, was undetectable on D23, and became normal on D77. Sperm with normal morphology reached 5.9 ± 2.6% on D35 but recovered (86.8 ± 2.1%) on D91. Sperm DNA integrity decreased from 86.5 ± 4.7% before insulation to 11.1 ± 3.7% on D63, returning to pre-insulation values on D120. Sperm BSP immunostaining was reduced after scrotal insulation. Variations in seminal protein abundances coincided with changes in sperm parameters. Seminal plasma superoxide dismutase, carboxypeptidase Q-precursor and NPC intracellular cholesterol transporter 2 decreased on D18, returning to normal after D28. Albumin, inhibitor of carbonic anhydrase precursor, EGF-like repeat and discoid I-like domain-containing protein 3 and polymeric immunoglobulin receptor increased after insulation. In summary, intermittent scrotal insulation drastically altered ram sperm attributes and seminal proteins, especially those associated with oxidative stress. Knowledge of animal´s response to thermal stress is vital in the scenario of climate changes.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140006222","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}
Pub Date : 2024-02-23DOI: 10.1016/j.anireprosci.2024.107436
G. Gebreyesus , J.B. Secher , M.S. Lund , K. Kupisiewicz , M. Ivask , T. Hallap , P. Pärn , G. Su
In vitro production of embryos (IVP) is increasingly applied in dairy cattle breeding and promises widespread use of females of superior genetic merits. One of the current challenges with implementation of IVP is the variability in blastocyst rates. Several factors contribute to these variabilities, among which is known to be the bull used for oocytes fertilization. The extent of genetic control of bulls’ effect on IVP performances is yet to be investigated. This study estimates genetic parameters for bull effects on IVP performance traits including blastocyst rate, hatching rate and an index trait combining Blastocyst rate, Kinetic Score, and Morphology score (BL_M_K). The IVP experiments were performed using oocytes aspirated from slaughterhouse ovaries from Holstein cows, fertilized with semen from 123 Holstein bulls. A total of 77 in vitro fertilization (IVF) experiments with 163 records (different IVF groups) were available for the analysis. The results indicate low to moderate heritability and moderate to high repeatability estimates for bull effects on IVP performance traits. Our study also showed that some semen quality traits had significant effects on IVP performance. This included strong genetic correlations between pre-cryopreservation sperm viability and blastocyst rate as well as BL_M_K score at days 7 and 8. Despite the generally weak bull effect correlations and the high standard errors of the estimates, our results provide initial evidence of a measurable genetic component in the bull's impact on IVP performance traits. However, the high standard errors underscore the need for further studies with a larger sample size.
{"title":"Genetic parameters for bull effects on in vitro embryo production (IVP) and relationship between semen quality traits and IVP performance","authors":"G. Gebreyesus , J.B. Secher , M.S. Lund , K. Kupisiewicz , M. Ivask , T. Hallap , P. Pärn , G. Su","doi":"10.1016/j.anireprosci.2024.107436","DOIUrl":"10.1016/j.anireprosci.2024.107436","url":null,"abstract":"<div><p><em>In vitro</em> production of embryos (IVP) is increasingly applied in dairy cattle breeding and promises widespread use of females of superior genetic merits. One of the current challenges with implementation of IVP is the variability in blastocyst rates. Several factors contribute to these variabilities, among which is known to be the bull used for oocytes fertilization. The extent of genetic control of bulls’ effect on IVP performances is yet to be investigated. This study estimates genetic parameters for bull effects on IVP performance traits including blastocyst rate, hatching rate and an index trait combining Blastocyst rate, Kinetic Score, and Morphology score (BL_M_K). The IVP experiments were performed using oocytes aspirated from slaughterhouse ovaries from Holstein cows, fertilized with semen from 123 Holstein bulls. A total of 77 <em>in vitro</em> fertilization (IVF) experiments with 163 records (different IVF groups) were available for the analysis. The results indicate low to moderate heritability and moderate to high repeatability estimates for bull effects on IVP performance traits. Our study also showed that some semen quality traits had significant effects on IVP performance. This included strong genetic correlations between pre-cryopreservation sperm viability and blastocyst rate as well as BL_M_K score at days 7 and 8. Despite the generally weak bull effect correlations and the high standard errors of the estimates, our results provide initial evidence of a measurable genetic component in the bull's impact on IVP performance traits. However, the high standard errors underscore the need for further studies with a larger sample size.</p></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2024-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0378432024000277/pdfft?md5=8db5ce2302afa8720feedec308fa92f0&pid=1-s2.0-S0378432024000277-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139951967","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}