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Ascorbic acid and resveratrol improve the structural integrity of the extracellular matrix and enhance follicular survival in cultured bovine ovarian tissue
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-22 DOI: 10.1016/j.theriogenology.2025.01.020
F.C. Costa , B.R. Silva , F.F.C. Filho , V.S. Bezerra , V.A.N. Azevedo , A.A. Silva , J.R.V. Silva
This study aimed to investigate the changes induced by the culture system and the effect of ascorbic acid and resveratrol on collagen fibers, stromal cells, follicle growth and survival, as well as antioxidant enzyme activity in cultured bovine ovarian tissues. In experiment 1, bovine ovarian fragments were cultured in α-minimum essential medium (α-MEM+) for 6 days. Before and after culturing, the fragments were fixed and processed to assess follicular morphology and diameters, stromal cell survival, collagen fibers, and glycosaminoglycans (GAGs). Uncultured and cultured tissues were also used to measure mRNA expression for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and peroxiredoxin (PRDX). Thiol levels and activity of CAT, SOD, and GPX enzymes were also investigated. In experiment 2, bovine ovarian fragments were cultured in α-MEM+ alone or supplemented with 50 μg/mL ascorbic acid or both 50 μg/mL ascorbic acid and 20 μM resveratrol for 6 days. In experiment 1, cultured tissues had higher percentages of growing follicles, but higher percentage of degenerated follicles than uncultured slices (P < 0.05). Additionally, the collagen and GAGs network became disorganized, with reduced deposition around primordial and primary follicles (P < 0.05). The number of stromal and granulosa cells, as well as follicular and oocyte diameters were reduced in both follicular categories compared to uncultured tissue (P < 0.05). Expression of mRNA for CAT, SOD, GPX, and PRDX was downregulated in 6-day cultured tissues (P < 0.05). Similarly, thiol levels and CAT activity were also reduced (P < 0.05). In experiment 2, ascorbic acid or both ascorbic acid and resveratrol increased the rate of follicular diameters and survival, and the number of granulosa and stromal cells compared to tissues cultured in the control medium (P < 0.05). Both ascorbic acid and resveratrol improved collagen density and preserved the GAG network, as well as increased thiol levels and CAT activity (P < 0.05). In conclusion, in vitro culture of ovarian tissue favored follicular activation, but reduced the proportion of normal follicles, collagen, GAG network, stromal cell numbers, and tissue antioxidant protection. Ascorbic acid alone or in association with resveratrol improved the preservation of extracellular matrix components and enhanced follicular survival.
{"title":"Ascorbic acid and resveratrol improve the structural integrity of the extracellular matrix and enhance follicular survival in cultured bovine ovarian tissue","authors":"F.C. Costa ,&nbsp;B.R. Silva ,&nbsp;F.F.C. Filho ,&nbsp;V.S. Bezerra ,&nbsp;V.A.N. Azevedo ,&nbsp;A.A. Silva ,&nbsp;J.R.V. Silva","doi":"10.1016/j.theriogenology.2025.01.020","DOIUrl":"10.1016/j.theriogenology.2025.01.020","url":null,"abstract":"<div><div>This study aimed to investigate the changes induced by the culture system and the effect of ascorbic acid and resveratrol on collagen fibers, stromal cells, follicle growth and survival, as well as antioxidant enzyme activity in cultured bovine ovarian tissues. In experiment 1, bovine ovarian fragments were cultured in α-minimum essential medium (α-MEM<sup>+</sup>) for 6 days. Before and after culturing, the fragments were fixed and processed to assess follicular morphology and diameters, stromal cell survival, collagen fibers, and glycosaminoglycans (GAGs). Uncultured and cultured tissues were also used to measure mRNA expression for superoxide dismutase (<em>SOD</em>), catalase (<em>CAT</em>), glutathione peroxidase (<em>GPX</em>), and peroxiredoxin (<em>PRDX</em>). Thiol levels and activity of CAT, SOD, and GPX enzymes were also investigated. In experiment 2, bovine ovarian fragments were cultured in α-MEM<sup>+</sup> alone or supplemented with 50 μg/mL ascorbic acid or both 50 μg/mL ascorbic acid and 20 μM resveratrol for 6 days. In experiment 1, cultured tissues had higher percentages of growing follicles, but higher percentage of degenerated follicles than uncultured slices (P &lt; 0.05). Additionally, the collagen and GAGs network became disorganized, with reduced deposition around primordial and primary follicles (P &lt; 0.05). The number of stromal and granulosa cells, as well as follicular and oocyte diameters were reduced in both follicular categories compared to uncultured tissue (P &lt; 0.05). Expression of mRNA for <em>CAT, SOD, GPX</em>, and <em>PRDX</em> was downregulated in 6-day cultured tissues (P &lt; 0.05). Similarly, thiol levels and CAT activity were also reduced (P &lt; 0.05). In experiment 2, ascorbic acid or both ascorbic acid and resveratrol increased the rate of follicular diameters and survival, and the number of granulosa and stromal cells compared to tissues cultured in the control medium (P &lt; 0.05). Both ascorbic acid and resveratrol improved collagen density and preserved the GAG network, as well as increased thiol levels and CAT activity (P &lt; 0.05). In conclusion, <em>in vitro</em> culture of ovarian tissue favored follicular activation, but reduced the proportion of normal follicles, collagen, GAG network, stromal cell numbers, and tissue antioxidant protection. Ascorbic acid alone or in association with resveratrol improved the preservation of extracellular matrix components and enhanced follicular survival.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 231-244"},"PeriodicalIF":2.4,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143060828","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
Cumulus cells and the TNF-alpha signaling facilitate aging of ovine oocytes
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-21 DOI: 10.1016/j.theriogenology.2025.01.019
Shuai Gong , Yan Zhang , Chao-Qun Cong, Guo-Liang Wang, Lin Jiang, Hong-Jie Yuan, Jing-He Tan, Ming-Jiu Luo
Post-maturation oocyte aging (PMOA) is known to significantly impair the developmental potential of oocytes; however, comprehensive studies on ovine PMOA remain limited. In mice, cumulus cells (CCs) accelerate oocyte aging by releasing cytokines, but the roles of CCs and cytokines in PMOA of domestic animals are poorly understood. This study aimed to elucidate the involvement of CCs and tumor necrosis factor (TNF)-α in the PMOA of ovine oocytes. Our findings reveal that PMOA significantly reduced blastocyst rates and the expression of development-promoting genes, while increasing oocyte degeneration and activation rates, along with expression of development-inhibiting genes, compared to newly matured oocytes. These detrimental effects were more pronounced in oocytes aged as cumulus-oocyte complexes than as cumulus-denuded oocytes. Additionally, PMOA led to increased apoptotic rates, TNF-α production, and TNF receptor 1 (TNFR1) expression in CCs, coupled with a significant reduction in the expression of anti-apoptotic genes. Mature oocytes expressed TNFR1, with levels decreasing significantly during PMOA. Importantly, the addition of the TNF-α antagonist Etanercept to the aging medium markedly improved parthenogenetic embryo development and the expression of competence-related genes, while mitigating CC apoptosis during PMOA of COCs. In conclusion, PMOA compromises developmental potential while heightening oocyte degeneration and activation sensitivity in ovine oocytes. Cumulus cells exacerbate PMOA through increased TNF-α signaling activity, highlighting the potential of TNF-α antagonists as therapeutic agents to counteract the deleterious effects of PMOA.
{"title":"Cumulus cells and the TNF-alpha signaling facilitate aging of ovine oocytes","authors":"Shuai Gong ,&nbsp;Yan Zhang ,&nbsp;Chao-Qun Cong,&nbsp;Guo-Liang Wang,&nbsp;Lin Jiang,&nbsp;Hong-Jie Yuan,&nbsp;Jing-He Tan,&nbsp;Ming-Jiu Luo","doi":"10.1016/j.theriogenology.2025.01.019","DOIUrl":"10.1016/j.theriogenology.2025.01.019","url":null,"abstract":"<div><div>Post-maturation oocyte aging (PMOA) is known to significantly impair the developmental potential of oocytes; however, comprehensive studies on ovine PMOA remain limited. In mice, <em>cumulus</em> cells (CCs) accelerate oocyte aging by releasing cytokines, but the roles of CCs and cytokines in PMOA of domestic animals are poorly understood. This study aimed to elucidate the involvement of CCs and tumor necrosis factor (TNF)-α in the PMOA of ovine oocytes. Our findings reveal that PMOA significantly reduced blastocyst rates and the expression of development-promoting genes, while increasing oocyte degeneration and activation rates, along with expression of development-inhibiting genes, compared to newly matured oocytes. These detrimental effects were more pronounced in oocytes aged as <em>cumulus</em>-oocyte complexes than as <em>cumulus</em>-denuded oocytes. Additionally, PMOA led to increased apoptotic rates, TNF-α production, and TNF receptor 1 (TNFR1) expression in CCs, coupled with a significant reduction in the expression of anti-apoptotic genes. Mature oocytes expressed TNFR1, with levels decreasing significantly during PMOA. Importantly, the addition of the TNF-α antagonist Etanercept to the aging medium markedly improved parthenogenetic embryo development and the expression of competence-related genes, while mitigating CC apoptosis during PMOA of COCs. In conclusion, PMOA compromises developmental potential while heightening oocyte degeneration and activation sensitivity in ovine oocytes. Cumulus cells exacerbate PMOA through increased TNF-α signaling activity, highlighting the potential of TNF-α antagonists as therapeutic agents to counteract the deleterious effects of PMOA.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 210-220"},"PeriodicalIF":2.4,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041428","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
Pseudopregnancy in dromedary camels: Characterization, prevalence, and clinical and hormonal properties
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-20 DOI: 10.1016/j.theriogenology.2025.01.010
Khalid S. Almushawwah , Derar R. Derar , Ahmed Ali
Pseudopregnancy is the development of signs of pregnancy in the absence of an embryo or fetus. The objectives of this study were to characterize pseudopregnancy in dromedary camels, determine its prevalence in camel farms and practice, identify associated risk factors, and describe its clinical and hormonal properties. The prevalence of pseudopregnancy on 100 camel farms with 4264 total female camels was determined to be 2.07 % (86/4264) overall, while the rate among infertile animals was 17.68 % (1385/7833). The genital tracts of 58 pseudopregnant female camels were examined, and their breeding histories were examined. Serum concentrations of estradiol-17 β (E2), progesterone (P4), and prolactin (PRL) in these animals were assessed. Five cyclic camels and five in early pregnancy were used as control subjects. Signs of pseudopregnancy included being anestrous, refusing to mate, and adopting a stiffened posture—with the head held high and the tail cocked—when approached by a male. Normal pregnancy-associated mammary and abdominal changes were absent. Risk factors associated with pseudopregnancy included age (odds ratio [OR] = 21.63, P = 0.0001) and a history of reproductive disorders (OR = 4.155, P = 0.042). Based on their P4 levels, the pseudopregnant camels were classified as either typical (high P4, 16/58, 27.59 %) or atypical (low P4, 42/58, 72.41 %). The main clinical findings in the camels with typical pseudopregnancies were a narrow/closed cervix (56.25 %), clinical endometritis (CE; 43.75 %), and pyometra (31.25 %), while those with atypical pseudopregnancies exhibited CE (50 %) and luteinized follicles (43.1 %). The pregnant camels had significantly (P = 0.0001) higher serum P4 concentrations (2.44 ± 0.32 ng/mL) than the pseudopregnant (0.68 ± 0.12 ng/mL) and cyclic camels (0.16 ± 0.01 ng/mL). Serum E2 levels did not differ significantly among the pseudopregnant (43.2 ± 1.05 pg/mL), pregnant (47.72 ± 4.06 pg/mL), and cyclic (40.72 ± 1.03 pg/mL) camels. The pregnant camels had a significantly (P = 0.04) higher average serum PRL concentration (3.61 ± 0.45 ng/mL) than the pseudopregnant (2.77 ± 0.12 ng/mL) and cyclic camels (2.18 ± 0.11). In conclusion, pseudopregnancy in camels exhibits the same external signs that characterize pregnancy, but pseudopregnancy involves an absence of edema of the udder, milk production, and high PRL levels. We propose a division of pseudopregnant camels into typical and atypical classes, depending on whether they have high P4 levels. Pseudopregnancy is associated with a high incidence of other reproductive disorders, particularly in older camels.
{"title":"Pseudopregnancy in dromedary camels: Characterization, prevalence, and clinical and hormonal properties","authors":"Khalid S. Almushawwah ,&nbsp;Derar R. Derar ,&nbsp;Ahmed Ali","doi":"10.1016/j.theriogenology.2025.01.010","DOIUrl":"10.1016/j.theriogenology.2025.01.010","url":null,"abstract":"<div><div>Pseudopregnancy is the development of signs of pregnancy in the absence of an embryo or fetus. The objectives of this study were to characterize pseudopregnancy in dromedary camels, determine its prevalence in camel farms and practice, identify associated risk factors, and describe its clinical and hormonal properties. The prevalence of pseudopregnancy on 100 camel farms with 4264 total female camels was determined to be 2.07 % (86/4264) overall, while the rate among infertile animals was 17.68 % (1385/7833). The genital tracts of 58 pseudopregnant female camels were examined, and their breeding histories were examined. Serum concentrations of estradiol-17 β (E2), progesterone (P4), and prolactin (PRL) in these animals were assessed. Five cyclic camels and five in early pregnancy were used as control subjects. Signs of pseudopregnancy included being anestrous, refusing to mate, and adopting a stiffened posture—with the head held high and the tail cocked—when approached by a male. Normal pregnancy-associated mammary and abdominal changes were absent. Risk factors associated with pseudopregnancy included age (odds ratio [OR] = 21.63, P = 0.0001) and a history of reproductive disorders (OR = 4.155, P = 0.042). Based on their P4 levels, the pseudopregnant camels were classified as either typical (high P4, 16/58, 27.59 %) or atypical (low P4, 42/58, 72.41 %). The main clinical findings in the camels with typical pseudopregnancies were a narrow/closed cervix (56.25 %), clinical endometritis (CE; 43.75 %), and pyometra (31.25 %), while those with atypical pseudopregnancies exhibited CE (50 %) and luteinized follicles (43.1 %). The pregnant camels had significantly (P = 0.0001) higher serum P4 concentrations (2.44 ± 0.32 ng/mL) than the pseudopregnant (0.68 ± 0.12 ng/mL) and cyclic camels (0.16 ± 0.01 ng/mL). Serum E2 levels did not differ significantly among the pseudopregnant (43.2 ± 1.05 pg/mL), pregnant (47.72 ± 4.06 pg/mL), and cyclic (40.72 ± 1.03 pg/mL) camels. The pregnant camels had a significantly (P = 0.04) higher average serum PRL concentration (3.61 ± 0.45 ng/mL) than the pseudopregnant (2.77 ± 0.12 ng/mL) and cyclic camels (2.18 ± 0.11). In conclusion, pseudopregnancy in camels exhibits the same external signs that characterize pregnancy, but pseudopregnancy involves an absence of edema of the udder, milk production, and high PRL levels. We propose a division of pseudopregnant camels into typical and atypical classes, depending on whether they have high P4 levels. Pseudopregnancy is associated with a high incidence of other reproductive disorders, particularly in older camels.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 196-202"},"PeriodicalIF":2.4,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041539","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
Reproduction control in a feral feline population using an anti-GnRH vaccine
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-19 DOI: 10.1016/j.theriogenology.2025.01.017
García María Florencia , Nuñez Favre Romina , Stornelli María Cecilia , Garcia Mitacek María Carla , Rearte Ramiro , de la Sota Rodolfo Luzbel , Stornelli María Alejandra
This study aimed to evaluate the use of the Improvac® vaccine to avoid heat and pregnancies in queens and fertility in males during the breeding season. Twenty-eight intact animals were divided into treated males (G1, n = 7), treated females (G2, n = 18), and untreated males (G3, n = 3) that were untamed and could not be captured for immunization. In cats from the G1 group, the testicular volume (337.35 ± 95.74 mm3) and testosterone concentration (1.31 ± 0.49 ng/mL) reached the lowest value 16 weeks after the first vaccination. At week 40, all queens exhibited both estrus cytology and estrus behavior, with serum estrogen (38.5 ± 1.93 pg/mL) and progesterone (0.5 ng/mL) concentrations within the physiological range for the phase. Eleven queens received a third dose of the vaccine at week 40, and none became pregnant by week 64. The remaining queens (n = 7) did not receive the third dose of the vaccine and became pregnant by week 44. In cats from the G1 and G2 groups, the hematologic parameters were within the physiological range for the species. The results of this study indicate that the Improvac® vaccine is safe and effective in the short to medium term in preventing cat reproduction.
{"title":"Reproduction control in a feral feline population using an anti-GnRH vaccine","authors":"García María Florencia ,&nbsp;Nuñez Favre Romina ,&nbsp;Stornelli María Cecilia ,&nbsp;Garcia Mitacek María Carla ,&nbsp;Rearte Ramiro ,&nbsp;de la Sota Rodolfo Luzbel ,&nbsp;Stornelli María Alejandra","doi":"10.1016/j.theriogenology.2025.01.017","DOIUrl":"10.1016/j.theriogenology.2025.01.017","url":null,"abstract":"<div><div>This study aimed to evaluate the use of the Improvac® vaccine to avoid heat and pregnancies in queens and fertility in males during the breeding season. Twenty-eight intact animals were divided into treated males (G1, n = 7), treated females (G2, n = 18), and untreated males (G3, n = 3) that were untamed and could not be captured for immunization. In cats from the G1 group, the testicular volume (337.35 ± 95.74 mm<sup>3</sup>) and testosterone concentration (1.31 ± 0.49 ng/mL) reached the lowest value 16 weeks after the first vaccination. At week 40, all queens exhibited both estrus cytology and estrus behavior, with serum estrogen (38.5 ± 1.93 pg/mL) and progesterone (0.5 ng/mL) concentrations within the physiological range for the phase. Eleven queens received a third dose of the vaccine at week 40, and none became pregnant by week 64. The remaining queens (n = 7) did not receive the third dose of the vaccine and became pregnant by week 44. In cats from the G1 and G2 groups, the hematologic parameters were within the physiological range for the species. The results of this study indicate that the Improvac® vaccine is safe and effective in the short to medium term in preventing cat reproduction.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 203-209"},"PeriodicalIF":2.4,"publicationDate":"2025-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041506","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
Metabolome and oxidative stress markers in the seminal plasma of Holstein bulls and their relationship with the characteristics of fresh and frozen/thawed sperm
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-17 DOI: 10.1016/j.theriogenology.2025.01.015
E. Giaretta , A. Damato , L. Zennaro , V. Bonfatti , B. Mislei , V. Vigolo , M.E. Falomo , F. Bertuzzo , G. Gabai , D. Bucci
Seminal plasma composition has important role in sperm functionality and its freezability. The objective of this study was to test the hypothesis that seminal plasma (SP) oxidative status and metabolome are associated with fresh semen characteristics and freezability of bull sperm. To accomplish this objective, oxidative status markers and metabolome of SP of ejaculates obtained from 20 Holstein bulls (3 for each bull) were analyzed using spectrophotometry and nuclear magnetic resonance (1H NMR). The ejaculates were classified into higher motility fresh semen (HMF) and lower motility fresh semen (LMF), according to total motility (TM) and progressive motility (PM) values of fresh semen. Then the ejaculates was cryopreserved and assigned to higher motility thawed group (HMT) or lower motility thawed group (LMT) according to TM and PM at 0 h post-thawing. Multivariate analyses were performed to identify the association between the functional characteristics of fresh and thawed semen and the SP parameters, in terms of the oxidative status and the metabolomic composition. According to our results, the advanced oxidative protein products (AOPP) and thiol concentrations in SP are significantly related to some physiological characteristics of the thawed sperm, such as higher viability, TM, PM and LIN and lower mitochondrial and cytoplasmic superoxide production in viable thawed cells. In contrast, a higher amount of C in the SP was negatively related to TM and PM of thawed semen and was associated with higher mitochondrial and cytoplasmic superoxide production.
In addition, partial least squares-discriminant analysis (PLS-DA) performed on the 1H NMR spectra indicated a discrete separation between HMF and LMF groups, and good discrimination between HMT and LMT groups. Higher levels of formic acid, lactate, glycerol and phosphocholine, were found in the SP of the HMF group than in the LMF group. On the other hand, alanine, phenylalanine, and tyrosine were higher in the SP of the LMF group than in the HMF group. GABA, glutamate, histidine and glycerol were found in higher concentrations in the HMT group than in the LMT group, while fructose decreased in the HMT group. Our results showed that the oxidative and metabolomic status of SP is related to the physiological properties of semen and its freezability and open new fields in research of SP biomarkers of bull semen preservation and fertility.
{"title":"Metabolome and oxidative stress markers in the seminal plasma of Holstein bulls and their relationship with the characteristics of fresh and frozen/thawed sperm","authors":"E. Giaretta ,&nbsp;A. Damato ,&nbsp;L. Zennaro ,&nbsp;V. Bonfatti ,&nbsp;B. Mislei ,&nbsp;V. Vigolo ,&nbsp;M.E. Falomo ,&nbsp;F. Bertuzzo ,&nbsp;G. Gabai ,&nbsp;D. Bucci","doi":"10.1016/j.theriogenology.2025.01.015","DOIUrl":"10.1016/j.theriogenology.2025.01.015","url":null,"abstract":"<div><div>Seminal plasma composition has important role in sperm functionality and its freezability. The objective of this study was to test the hypothesis that seminal plasma (SP) oxidative status and metabolome are associated with fresh semen characteristics and freezability of bull sperm. To accomplish this objective, oxidative status markers and metabolome of SP of ejaculates obtained from 20 Holstein bulls (3 for each bull) were analyzed using spectrophotometry and nuclear magnetic resonance (<sup>1</sup>H NMR). The ejaculates were classified into higher motility fresh semen (HMF) and lower motility fresh semen (LMF), according to total motility (TM) and progressive motility (PM) values of fresh semen. Then the ejaculates was cryopreserved and assigned to higher motility thawed group (HMT) or lower motility thawed group (LMT) according to TM and PM at 0 h post-thawing. Multivariate analyses were performed to identify the association between the functional characteristics of fresh and thawed semen and the SP parameters, in terms of the oxidative status and the metabolomic composition. According to our results, the advanced oxidative protein products (AOPP) and thiol concentrations in SP are significantly related to some physiological characteristics of the thawed sperm, such as higher viability, TM, PM and LIN and lower mitochondrial and cytoplasmic superoxide production in viable thawed cells. In contrast, a higher amount of C in the SP was negatively related to TM and PM of thawed semen and was associated with higher mitochondrial and cytoplasmic superoxide production.</div><div>In addition, partial least squares-discriminant analysis (PLS-DA) performed on the <sup>1</sup>H NMR spectra indicated a discrete separation between HMF and LMF groups, and good discrimination between HMT and LMT groups. Higher levels of formic acid, lactate, glycerol and phosphocholine, were found in the SP of the HMF group than in the LMF group. On the other hand, alanine, phenylalanine, and tyrosine were higher in the SP of the LMF group than in the HMF group. GABA, glutamate, histidine and glycerol were found in higher concentrations in the HMT group than in the LMT group, while fructose decreased in the HMT group. Our results showed that the oxidative and metabolomic status of SP is related to the physiological properties of semen and its freezability and open new fields in research of SP biomarkers of bull semen preservation and fertility.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 262-274"},"PeriodicalIF":2.4,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143075572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Porcine endometrial epithelial organoids: Generation, characterization, and the impact of stromal cells
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-17 DOI: 10.1016/j.theriogenology.2025.01.013
Meng-Die Li, Hai-Yue Xu, Yu-Shi Chen, Xin-Cheng Zhang, Hua Ni
Endometrial organoids (EOs) are three-dimensional models that emulate the endometrium, serving as an invaluable in vitro tool for investigating the cellular and molecular mechanisms underlying endometrial physiology and pathology during the estrous cycle and pregnancy. While significant progress has been made in the establishment and optimization of EOs for both humans and mice, research on such models in other species remains limited. This study aimed to develop porcine endometrial epithelial organoids (EEOs) to explore the regulatory mechanisms of uterine function and maternal-fetal interactions during porcine pregnancy, which are critical for enhancing reproductive efficiency and improving embryo transfer techniques. Utilizing a defined protocol involving Matrigel and a simplified culture medium, we successfully established EEOs from porcine endometrial epithelial cells, which demonstrated epithelial characteristics, proliferative capacity, glycogen secretion, and hormonal responsiveness. Co-culturing with porcine endometrial stromal cells not only enhanced EEO formation but also mitigated the negative effect of progesterone on glycogen secretion. This cost-effective and efficient method provides a valuable reference for EO generation in other large animal species.
{"title":"Porcine endometrial epithelial organoids: Generation, characterization, and the impact of stromal cells","authors":"Meng-Die Li,&nbsp;Hai-Yue Xu,&nbsp;Yu-Shi Chen,&nbsp;Xin-Cheng Zhang,&nbsp;Hua Ni","doi":"10.1016/j.theriogenology.2025.01.013","DOIUrl":"10.1016/j.theriogenology.2025.01.013","url":null,"abstract":"<div><div>Endometrial organoids (EOs) are three-dimensional models that emulate the endometrium, serving as an invaluable <em>in vitro</em> tool for investigating the cellular and molecular mechanisms underlying endometrial physiology and pathology during the estrous cycle and pregnancy. While significant progress has been made in the establishment and optimization of EOs for both humans and mice, research on such models in other species remains limited. This study aimed to develop porcine endometrial epithelial organoids (EEOs) to explore the regulatory mechanisms of uterine function and maternal-fetal interactions during porcine pregnancy, which are critical for enhancing reproductive efficiency and improving embryo transfer techniques. Utilizing a defined protocol involving Matrigel and a simplified culture medium, we successfully established EEOs from porcine endometrial epithelial cells, which demonstrated epithelial characteristics, proliferative capacity, glycogen secretion, and hormonal responsiveness. Co-culturing with porcine endometrial stromal cells not only enhanced EEO formation but also mitigated the negative effect of progesterone on glycogen secretion. This cost-effective and efficient method provides a valuable reference for EO generation in other large animal species.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 175-183"},"PeriodicalIF":2.4,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Supplementation with N-Acetyl-L-cysteine during in vitro maturation improves goat oocyte developmental competence by regulating oxidative stress
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-17 DOI: 10.1016/j.theriogenology.2025.01.016
Yanfei Wang , Qingwei Wang , Quan Ji , Pengcheng An , Xiaodong Wang , Yonghong Ju , Ruiyang Li , Yong Ruan , Jiafu Zhao , Maosheng Cao , Xiang Chen
Oocyte quality can affect mammal fertilization rate, early embryonic development, pregnancy maintenance, and fetal development. Oxidative stress induced by reactive oxygen species (ROS) is one of the most important causes of poor oocyte maturation in vitro. To reduce the degree of cellular damage caused by ROS, supplementation with the antioxidant N-Acetyl-L-cysteine (NAC) serves as an effective pathway to alleviate glutathione (GSH) depletion during oxidative stress. This study investigated the effects of NAC supplementation during in vitro maturation of goat oocytes and explored the molecular mechanisms of maturation by transcriptome sequencing of MⅡ oocytes. The results showed that 1.5 mM NAC significantly increased the rates of oocyte maturation and cumulus cell expansion and improved the subsequent development of embryos. During the subsequent culture of parthenogenetically activated embryos, 1.5 mM NAC significantly increased the division rate of oocytes and blastocyst rate. It also reduced the accumulation of ROS, increased the level of GSH, alleviated oxidative stress, and enhanced the antioxidant capacity and cell metabolic activity. Transcriptome sequencing results revealed that NAC treatment significantly increased the expression of SIRT1, GGCT, and MITF genes related to the cellular antioxidant system, as well as the IDH3G gene related to energy metabolism, and decreased the expression of CASP8, FOS, and MMP1 genes related to apoptosis and cell invasion, as well as the CCL2. and CXCL8 genes related to the inflammatory response. In conclusion, the findings suggest that NAC supplementation significantly reduces oxidative stress, improves antioxidant capacity and metabolic activity, promotes oocyte maturation, and improves oocyte quality.
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引用次数: 0
miRNA-125a regulates porcine oocyte maturation in vitro by targeting ADAR
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-15 DOI: 10.1016/j.theriogenology.2025.01.011
Yue Han , Ping Lu , Yongsheng Yu , Weiyu Gu , Chunyu Li , Yanqiu Lv , Xinglin Qu , Yuyang Zhang , Qinglong Xu , Shunfa Yao , Xuan Chen , Yi Jin
Follicular fluid extracellular vesicles are beneficial for in vitro oocyte maturation and development; however, their effect on the expression profiles of oocyte microRNAs (miRNAs) and the roles of related miRNAs are unknown. In this study, we aimed to investigate miRNA expression in mature oocytes cultured in follicular fluid extracellular vesicles and the effect of miRNA-125a (miR-125a) on oocyte maturation. The expression profiles of the miRNAs were determined by microRNA sequencing, followed by target gene prediction analysis. We transfected miR-125a mimics and an miR-125a inhibitor to evaluate the effect of modulated miRNA-125a on cumulus expansion, oocyte maturation rate, changes in cytoplasmic maturation-related indicators, and changes in the expression of oocyte maturation-related, cumulus expansion-related, and predicted target genes. We found that miR-125a overexpression decreased the levels of cumulus expansion-related, oocyte maturation-related, and predicted target genes, adenosine deaminase RNA specific (ADAR), and lipid droplet number, and it increased the percentage of oocytes with abnormal cortical granule distribution. Inhibiting miR-125a increased the expression levels of oocyte maturation-related and target genes, number of lipid droplets, and endoplasmic reticulum function, and it decreased lipid droplet size. Mitochondrial membrane potential and reactive oxygen species levels were not significantly different between groups. In conclusion, our results suggest that extracellular vesicles may improve oocyte quality by modulating ADAR through regulating miR-125a.
{"title":"miRNA-125a regulates porcine oocyte maturation in vitro by targeting ADAR","authors":"Yue Han ,&nbsp;Ping Lu ,&nbsp;Yongsheng Yu ,&nbsp;Weiyu Gu ,&nbsp;Chunyu Li ,&nbsp;Yanqiu Lv ,&nbsp;Xinglin Qu ,&nbsp;Yuyang Zhang ,&nbsp;Qinglong Xu ,&nbsp;Shunfa Yao ,&nbsp;Xuan Chen ,&nbsp;Yi Jin","doi":"10.1016/j.theriogenology.2025.01.011","DOIUrl":"10.1016/j.theriogenology.2025.01.011","url":null,"abstract":"<div><div>Follicular fluid extracellular vesicles are beneficial for in vitro oocyte maturation and development; however, their effect on the expression profiles of oocyte microRNAs (miRNAs) and the roles of related miRNAs are unknown. In this study, we aimed to investigate miRNA expression in mature oocytes cultured in follicular fluid extracellular vesicles and the effect of miRNA-125a (miR-125a) on oocyte maturation. The expression profiles of the miRNAs were determined by microRNA sequencing, followed by target gene prediction analysis. We transfected miR-125a mimics and an miR-125a inhibitor to evaluate the effect of modulated miRNA-125a on cumulus expansion, oocyte maturation rate, changes in cytoplasmic maturation-related indicators, and changes in the expression of oocyte maturation-related, cumulus expansion-related, and predicted target genes. We found that miR-125a overexpression decreased the levels of cumulus expansion-related, oocyte maturation-related, and predicted target genes, adenosine deaminase RNA specific <em>(ADAR</em>), and lipid droplet number, and it increased the percentage of oocytes with abnormal cortical granule distribution. Inhibiting miR-125a increased the expression levels of oocyte maturation-related and target genes, number of lipid droplets, and endoplasmic reticulum function, and it decreased lipid droplet size. Mitochondrial membrane potential and reactive oxygen species levels were not significantly different between groups. In conclusion, our results suggest that extracellular vesicles may improve oocyte quality by modulating <em>ADAR</em> through regulating miR-125a.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"235 ","pages":"Pages 184-193"},"PeriodicalIF":2.4,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143024770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Strategic use of estrus intensity to combine timed artificial insemination and embryo transfer in large-scale cattle reproduction programs 在大规模牛繁殖计划中策略性地利用发情强度结合定时人工授精和胚胎移植。
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-13 DOI: 10.1016/j.theriogenology.2025.01.009
Fábio Morotti , Gustavo Martins Gomes dos Santos , Katia Cristina Silva-Santos , José Henrique Ayres Dias , Marcelo Marcondes Seneda
This study compared the reproductive performance of three different programs using conventional fixed-time artificial insemination (FTAI), fixed-time embryo transfer (FTET), and their combined use, considering estrus intensity as a criterion for the reproductive program. Brangus multiparous cows (n = 1.100), 40–50 days postpartum, 4–8 years old, and body condition scores 2.50 of 4.50 underwent a protocol for ovulation synchronization based on progesterone (P4) and estradiol on D0. On D10, one of three Programs I-III was implemented: control FTAI (n = 147 cows), control FTET (n = 617) with embryos from in vitro production (IVP) on D17, or FTAI + FTET (n = 336), where cows with low or no estrus expression were inseminated on D10, similar to Program I, while those with high-intensity expression received one embryo from IVP on D17, similar to Program II. Corpus luteum (CL) size and quality were assessed using B-mode and Doppler ultrasound on D17. The service/utilization rate was higher for FTAI and combined FTAI + FTET than for the conventional FTET. The program that used only FTAI resulted in higher pregnancy at 30 and 60 days, as well as lower pregnancy loss, compared to programs that used embryos. Furthermore, the FTAI + FTET program showed reduced pregnancy loss, compared to the FTET program. In the combined program, the CL was greater in those who received FTET alone than in those who received FTAI alone. A higher proportion of recipients with CL with a high luteal blood perfusion score was observed in the FTET group compared to the FTAI group. Satisfactory reproductive rates can be achieved using conventional FTAI or FTET programs. However, the combined program, associated with monitoring the intensity of estrus expression, is a promising strategy for allocating females with absent or low estrus expression to FTAI, and those with high estrus intensity to FTET. Recipients with high-intensity expression had higher CL quality and, when transferred, maintained satisfactory reproductive performance compared with conventional FTET.
本研究比较了常规定时人工授精(FTAI)、定时胚胎移植(fet)以及两者联合使用三种不同方案的生殖性能,并将发情强度作为生殖方案的评判标准。产后40 ~ 50天、4 ~ 8岁、体况评分为2.50 ~ 4.50的布兰格斯(Brangus)奶牛(n = 1.100)在D0采用基于黄体酮(P4)和雌二醇的排卵同步方案。在D10,实施了三个方案中的一个:对照FTAI (n = 147头),对照FTAI (n = 617头)与体外生产(IVP)胚胎在D17上的对照FTAI (n = 617头),或FTAI + FTET (n = 336头),其中低发情或没有发情表达的奶牛在D10上受精,类似于方案一,而高表达的奶牛在D17上接受一个IVP胚胎,类似于方案二。采用b超和多普勒超声对D17黄体大小和质量进行评价。与传统的fet相比,FTAI和FTAI + fet组合的服务/利用率更高。与使用胚胎的计划相比,仅使用FTAI的计划导致30天和60天的怀孕率更高,并且怀孕损失更低。此外,与fet计划相比,FTAI + fet计划显示减少了妊娠损失。在联合治疗方案中,单独接受fet治疗的患者的CL大于单独接受FTAI治疗的患者。与FTAI组相比,FTET组中具有高黄体血流灌注评分的CL受体比例更高。使用传统的FTAI或fet程序可以达到令人满意的生殖率。然而,结合监测发情表达强度的联合程序是一种很有前景的策略,可以将没有或低发情表达的雌性分配给FTAI,而将发情强度高的雌性分配给FTET。与传统fet相比,高强度表达的受体具有更高的CL质量,并且在转移时保持了令人满意的生殖性能。
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引用次数: 0
The occurrence of follicular cyst affects the embryonic developmental competences of buffalo oocytes under in vitro culture conditions 在体外培养条件下,卵泡囊肿的发生影响水牛卵母细胞的胚胎发育能力。
IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Pub Date : 2025-01-13 DOI: 10.1016/j.theriogenology.2025.01.007
Pradeep Saini , Sandeep Kumar , Hitesh Jangra , Anupama Solanki , Rahul Verma , Rajesh Kumar , Ramakant Kaushik , Kushal Grakh , Gitesh Saini , Yogesh Bangar , Anand Kumar Pandey
Cystic ovarian disease (COD) is a major cause of infertility in dairy cows. This study aimed to investigate the impact of follicular cysts on the invitro blastocyst developmental competence of oocytes and the relative gene expression of blastocysts developed from the subordinate follicles of ipsilateral (ovary with cyst), contralateral (ovary opposite to cyst), and normal ovaries of buffaloes. A total of 2059 ovaries were collected from slaughterhouse and classified into three categories based on the presence of follicular cysts: a) ipsilateral, b) contralateral, and c) control (absence of cysts). Oocytes of grades A, B, and C were used for invitro maturation, invitro fertilization, and invitro culture. The cleavage rates of the ipsilateral (91.54 %) and contralateral (95.71 %) categories were higher (P < 0.01) than those of the control (76.82 %). Conversely, the blastocyst development rate was higher (P < 0.01) in control (32.67 %) than in ipsilateral (8.46 %) and contralateral (7.14 %) groups. The mRNA expression levels of maturation (BMP15), steroidogenesis (STAR, CYP19), antioxidant (SOD2, HSPB1) and anti-apoptotic (BCL2) genes were decreased in arrested embryos derived from both cystic (ipsilateral and contralateral) and control category follicles when compared to blastocysts derived from their respective category follicles. Conversely, BAX expression increased in arrested embryos. Expression of SOD2 and BAX was downregulated in blastocysts from both ipsilateral and contralateral categories compared to controls. The presence of one or more follicular cysts in either ovary affected the developmental competence of oocytes derived from subordinate follicles. Therefore, the buffaloes with cysts in either ovary should be avoided when aspirating follicles for IVF.
卵巢囊性疾病(COD)是奶牛不育的主要原因。本研究旨在探讨卵泡囊肿对水牛同侧卵巢(有囊肿的卵巢)、对侧卵巢(与囊肿相对的卵巢)和正常卵巢的卵泡发育能力的影响以及囊胚的相关基因表达。从屠宰场采集了2059个卵巢,并根据卵泡囊肿的存在将其分为三类:A)同侧,b)对侧和c)对照(未见囊肿)。A级、B级和C级卵母细胞用于体外成熟、体外受精和体外培养。同侧(91.54%)和对侧(95.71%)类型的卵裂率较高(P
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引用次数: 0
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Theriogenology
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