TheriogenologyPub Date : 2024-09-10DOI: 10.1016/j.theriogenology.2024.09.006
{"title":"MAT2A is essential for zygotic genome activation by maintaining of histone methylation in porcine embryos","authors":"","doi":"10.1016/j.theriogenology.2024.09.006","DOIUrl":"10.1016/j.theriogenology.2024.09.006","url":null,"abstract":"<div><p>Methionine adenosyltransferase 2A (MAT2A) is an essential enzyme in the methionine cycle that generates S-adenosylmethionine (SAM) by reacting with methionine and ATP. SAM acts as a methyl donors for histone and DNA methylation, which plays key roles in zygotic genome activation (ZGA). However, the effects of MAT2A on porcine ZGA remain unclear. To investigate the function of MAT2A and its underlying mechanism in porcine ZGA, MAT2A was knocked down by double-stranded RNA injection at the 1-cell stage. MAT2A is highly expressed at every stage of porcine embryo development. The percentages of four-cell-stage embryos and blastocysts were lower in the MAT2A-knockdown (KD) group than in the control group. Notably, depletion of MAT2A decreased the levels of H3K4me2, H3K9me2/3, and H3K27me3 at the four-cell stage, whereas MAT2A KD reduced the transcriptional activity of ZGA genes. MAT2A KD decreased embryonic ectoderm development (EED) and enhancer of zeste homolog 2 (EZH2) expression. Exogenous SAM supplementation rescued histone methylation levels and developmental arrest induced by MAT2A KD. Additionally, MAT2A KD significantly increased DNA damage and apoptosis. In conclusion, MAT2A is involved in regulating transcriptional activity and is essential for regulating histone methylation during porcine ZGA.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142172551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-10DOI: 10.1016/j.theriogenology.2024.09.009
{"title":"Pentose phosphate pathway inhibition during in vitro maturation substantially affects the metabolism of bovine COCs and blastocysts","authors":"","doi":"10.1016/j.theriogenology.2024.09.009","DOIUrl":"10.1016/j.theriogenology.2024.09.009","url":null,"abstract":"<div><p>Glucose metabolism is widely examined in terms of its effect on oocytes and embryos quality. There are two main pathways of glucose metabolism – glycolysis and pentose phosphate pathway (PPP). The glycolytic pathway allows for energy production in the form of ATP and metabolites such as pyruvate and lactate, whereas PPP activity generates NADPH as well as ribose 5-phosphate, a precursor for the synthesis of nucleotides. The aim of the present experiment was the selective inhibition of either glycolysis or PPP during <em>in vitro</em> maturation of bovine cumulus-oocyte complexes (COCs) to demonstrate, how it affects COCs and further embryos with regard to selected lipidomic and metabolomic aspects. Inhibitors of glycolysis (IO) or PPP (DHEA) were applied during IVM, and the control group was matured under standard conditions. A set of COCs from each group was fertilized and obtained embryos were cultured to the blastocyst stage. ATP level was measured in oocytes, relative mRNA level of selected genes involved in energy metabolism was measured in cumulus cells (CC; real time PCR), lipid droplets parameters were evaluated in oocytes and CC whereas metabolome and lipidome (mass spectrometry) were evaluated in oocytes, CC and blastocysts as well. The experiment shows that glycolysis inhibition during IVM affects mainly CC with no effect in oocytes. It allows to maintain the good developmental potential of oocytes and no negative effect of blastocysts quality and quantity is observed. In contrary, PPP inhibition negatively affects metabolic and lipidomic parameters of both oocyte and CC, which further decreases blastocyst rate and quality. It is therefore concluded that PPP is the most crucial pathway of glucose metabolism for COC developmental potential.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0093691X24003789/pdfft?md5=0015010f9e5293481dd8c95bab6be1d7&pid=1-s2.0-S0093691X24003789-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142172554","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-10DOI: 10.1016/j.theriogenology.2024.09.003
{"title":"The influence of lactation on insulin and glucose metabolism, lipid profile, and cytokines in pregnant mares","authors":"","doi":"10.1016/j.theriogenology.2024.09.003","DOIUrl":"10.1016/j.theriogenology.2024.09.003","url":null,"abstract":"<div><p>Glucose metabolism adapts to gestation, resulting in progressive physiological insulin resistance and increased insulin secretion to maintain maternal euglycemia and glucose availability for the developing fetus. These changes can impact mare fertility and maternal and neonatal health. This is the first comparison of body condition, regional adiposity, insulin and glucose dynamics, lipid metabolism, and cytokine production between lactating and non-lactating mares before, during pregnancy, and early postpartum. Twelve pregnancies from 9 broodmares, five nonlactating (NL) and seven lactating (L), were used. Evaluations were performed on the day of ovulation, at 55, 110, 165, 220, 275, and 330 days of gestation (D55, D110, D165, D220, D275, D330) and 21 days postpartum (21pp). Mares in the L group had lower basal insulin and glucose at the beginning of pregnancy, smaller area under the curve of insulin and glucose, and greater insulin sensitivity and glucose tolerance. Resistin was higher in D110 and D165 than in D0, D275, 330 and 21pp, while leptin was higher in D55, and in D110, at D110 it was equal to D0, D220, and D275, but higher than at D330 and D21pp. As for the groups, L presented lower body condition score (BCS), crest neck score (CNS), rump fat thickness (RUM), basal insulin, glucose area under the curve (AUCg), MIRG and higher RISQI, adiponectin and tumor necrosis factor (TNFα). There was no effect over time in non-esterified fatty acids (NEFA) concentrations between the L mares; in the NL, D275 presented higher concentrations than those of D0, D55, and D110, which in turn were equal to the other time points; there were higher concentrations in NL mares than L in samples D165 and D275. In conclusion, a different metabolic profile during pregnancy was detected, and NL mares were closer to the metabolic threshold for the occurrence of metabolic syndrome during pregnancy. Understanding the impacts of these differences on mare's health and their offspring's future is fundamental as most of our recipient mares for embryo transfer are non-lactating. Therefore, we suggest that further studies be performed to evaluate lactation's influence on mares' metabolic parameters.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142243911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-10DOI: 10.1016/j.theriogenology.2024.09.007
{"title":"miR-9-5p regulates Sirt1 involved in testicular development and spermatogenesis in mouse","authors":"","doi":"10.1016/j.theriogenology.2024.09.007","DOIUrl":"10.1016/j.theriogenology.2024.09.007","url":null,"abstract":"<div><p>Testicular development and spermatogenesis are critical for male reproduction, with histone (de)acetylation playing a key role in chromatin remodeling within germ cells. Sirt1, a key histone deacetylase, is implicated in chromatin remodeling, but its expression pattern and specific role in testicular development and spermatogenesis need further study. This study comprehensively analyzed Sirt1 expression in adult and juvenile mouse testicular tissues and across various male germ cells, utilizing RT-qPCR, Western blot, immunofluorescence, and cell transfection. GO and KEGG enrichment analyses were performed to elucidate the biological functions and pathways associated with <em>Sirt1</em> and its related genes. Multiple miRNA databases were utilized to predict miRNAs targeting <em>Sirt1</em>, and their expression levels were validated using RT-qPCR. Lentiviral transfection was used to knockdown candidate miRNAs to assess their functional roles. The results revealed a significant downregulation of Sirt1 expression in adult mouse testicular tissues compared to juvenile tissues, with pronounced variation across diverse male germ cells. Sirt1 was highly expressed in spermatogonia and mature sperm, but comparatively lower in spermatocytes and spermatids. GO and KEGG enrichment analyses highlighted Sirt1's role in key biological processes, including chromatin organization, regulation of cell proliferation, and energy homeostasis, as well as its association with signaling pathways like cellular senescence, the FoxO signaling pathway, and the AMPK signaling pathway. Bioinformatic analysis and subsequent RT-qPCR validation identified miR-9-5p as a miRNA targeting <em>Sirt1</em>. The expression of miR-9-5p was significantly higher in adult mouse testicular tissues compared to juvenile tissues, inversely correlating with Sirt1 levels. Moreover, the knockdown of miR-9-5p led to a notable increase in Sirt1 mRNA and protein expression. In conclusion, Sirt1 is a key player in mouse testicular development and spermatogenesis. The discovery that miR-9-5p negatively regulates <em>Sirt1</em> suggests a critical regulatory axis that may govern these processes, providing novel insights into male fertility and potential targets for therapeutic intervention.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142172553","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-06DOI: 10.1016/j.theriogenology.2024.09.005
{"title":"Evaluation of the productive and reproductive performance of sexual precocity at different ages in Nellore heifers","authors":"","doi":"10.1016/j.theriogenology.2024.09.005","DOIUrl":"10.1016/j.theriogenology.2024.09.005","url":null,"abstract":"<div><p>This study aimed to investigate the differences between productive and reproductive performance traits of sexually precocious and non-sexually precocious Nellore heifers and to evaluate the genetic correlation of sexual precocity with traits of economic importance. For this purpose, 300,000 Nellore heifers were evaluated for reproductive traits: heifer pregnancy (HP) at 14 (HP<sub>14</sub>), 18 (HP<sub>18</sub>), and 24 (HP<sub>24</sub>) months; heifer rebreeding (HR); number of progenies up to 53 months (NP<sub>53</sub>); and probability of the cow remaining in the herd until 76 months with at least 3 progenies (Stay). The growth-related traits evaluated included female yearling weight (YW); average daily gain from weaning to yearling (ADG<sub>W-Y</sub>); weight at maturity (MW); weaning weight of first progeny (WW<sub>prog</sub>); and female visual scores at yearling for conformation (Conf), precocity (Prec) and muscling (Musc). The effects of female YW and ADG<sub>W-Y</sub> in six categories on HP<sub>14</sub>, HP<sub>18</sub>, and HP<sub>24</sub> were analyzed using Generalized linear mixed models (GLMM). Furthermore, a linear mixed model was used to evaluate the impact of HP on WW<sub>prog</sub>, MW, and reproductive performance (NP<sub>53</sub> and Stay). Genetic correlations of HP evaluated in different months with growth and reproductive traits were estimated using a bivariate animal model. Precocious heifers (HP<sub>14</sub>) were lighter for YW and MW but had greater ADG<sub>W-Y</sub> than HP<sub>18</sub> and HP<sub>24</sub>. The probability for HP<sub>14</sub>, HP<sub>18</sub>, and HP<sub>24</sub> increased as the classes of YW and ADG<sub>W-Y</sub> increased. However, heifers weighing more than 326 kg had a slight reduction in the probability of becoming pregnant at HP<sub>14</sub> and HP<sub>18</sub>. Precocious heifers (HP<sub>14</sub> and HP<sub>18</sub>) produced their first progeny by 3 % lighter than HP<sub>24</sub>, although they had a greater NP<sub>53</sub>. Precocious heifers at 18 months (HP<sub>18</sub>) were 3 % and 6.8 % more likely to remain in the herd than HP<sub>14</sub> and HP<sub>24</sub> heifers, respectively. Genetic correlations between growth traits (WW, YW, ADG<sub>W-Y,</sub> and MW) and heifer pregnancy (HP<sub>14</sub>, HP<sub>18</sub>, and HP<sub>24</sub>) ranged from weak (r<sub>g</sub> = 0.27 ± 0.05) to moderate (r<sub>g</sub> = −0.47 ± 0.07). The genetic correlation between HR and HP was stronger for HP<sub>24</sub> (0.75) against HP<sub>14</sub> (0.58) and HP<sub>18</sub> (0.64). Although, the genetic correlation between NP<sub>53</sub> and Stay with HP<sub>14</sub> was higher (r<sub>g</sub> = 0.53 and 0.45) than those observed for HP<sub>18</sub> (r<sub>g</sub> = 0.46 and 0.38) and HP<sub>24</sub> (r<sub>g</sub> = 0.35 and 0.39). The genetic correlation estimates between HP and visual scores were moderate and favorable for HP<sub>14</sub>. Selecting HP<sub>14</sub> is beneficial for production ","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142243916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-06DOI: 10.1016/j.theriogenology.2024.09.002
{"title":"Anti-Müllerian hormone in feline cryptorchidism: Serum levels, tissue expression, and implications for testicular health","authors":"","doi":"10.1016/j.theriogenology.2024.09.002","DOIUrl":"10.1016/j.theriogenology.2024.09.002","url":null,"abstract":"<div><p>Anti-Müllerian hormone (AMH) has become a pivotal subject in the study of testicular descent, maturation, integrity, and male fertility. Recent studies explored its roles and implications across various domestic species. A prominent approach involved the understanding of the modulation of AMH in reproductive disorders, including cryptorchidism. While substantial findings have been reported in dogs, ruminants, swine, and horses, data on AMH in feline cryptorchidism remains limited. Here, we aimed to bridge this gap by comparing AMH serum levels among cryptorchid, healthy intact, and castrated tomcats, employing an enzyme-linked immunosorbent assay (ELISA) kit for quantification. In addition, AMH expression in retained and descended testes was evaluated through immunohistochemistry, with positive staining quantified via pixel analysis in two distinct regions of interest: the seminiferous tubule and the interstitial space. Furthermore, tissue samples were subjected to histological evaluation and morphometric analysis, which included the calculation of seminiferous tubule areas (STA) and assessment of Johnsen scores. Thus, the relationship between AMH expression, altered testicular histology, and impaired spermatogenesis could be examined. The expression of AMH in retained and descended testes, was investigated, and the relationship between AMH expression, altered testicular histology, and impaired spermatogenesis was examined. Mean serum AMH levels differed significantly (P < 0.001) across the different groups being 15.35 ± 4.66 ng/mL (mean ± SD) in healthy intact tomcats (n = 15), 25.55 ± 2.86 ng/mL (mean ± SD) in cryptorchids (n = 10) and below 0.015 ng/mL in castrated cats (n = 10). STAs and Johnsen scores were significantly reduced in retained testes when compared to descended gonads (P < 0.01). Furthermore, serum AMH was negatively correlated with both the STA (ρ = −0.725, P < 0.001) and the Johnsen scores (ρ = −0.699, P < 0.001), suggesting its potential value for tissue integrity and spermatogenesis evaluation. In addition, positive immunostaining was significantly higher in retained testes (P < 0.05), especially in the interstitial space (P < 0.01), suggesting an involvement of the Leydig cells. Additionally, the increased interstitial expression was linked to the degree of tissue degeneration and the impaired spermatogenesis being negatively corelated with both Johnsen scores (ρ = −0.309, P < 0.01) and STA (ρ = −0.208, P < 0.05). Our findings underscore the potential of AMH in assessing testicular health and reveal possible interspecific differences, stressing the need for further investigation in cats.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142168249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-04DOI: 10.1016/j.theriogenology.2024.09.004
{"title":"Analytical and clinical validation of diagnostic tests for the detection of leucospermia in beef bulls","authors":"","doi":"10.1016/j.theriogenology.2024.09.004","DOIUrl":"10.1016/j.theriogenology.2024.09.004","url":null,"abstract":"<div><p>The objectives of this study were to validate diagnostic tests to detect polymorphonuclear cells (PMNs) in bull semen, and to determine the prevalence of leucospermia in beef bulls with varying semen quality. We hypothesized that all tests have comparable diagnostic value, and that leucospermia is more prevalent in unsatisfactory breeders in association with poor semen quality. For the analytical validation, one ejaculate was obtained from five bulls. Aliquots of 50 × 10<sup>6</sup> purified sperm were incubated in triplicate with six concentrations of purified bovine PMNs: 1) no PMNs, 2) 0.25 × 10<sup>6</sup> PMN/ml, 3) 0.5 × 10<sup>6</sup> PMN/ml, 4) 2.5 × 10<sup>6</sup> PMN/ml, 5) 5 × 10<sup>6</sup> PMN/ml, 6) 10 × 10<sup>6</sup> PMN/ml. The PMNs were quantified using a hemacytometer, cytology, a leucocyte esterase dipstick test (LEDT), a peroxidase test, and CD45 immunolabeling. The number of leucocytes detected with the LEDT differed among treatments (P < 0.0001). The quantitative tests detected differences with the control treatment at a PMN concentration of ≥2.5 × 10<sup>6</sup> PMN/ml (P < 0.0001). Sperm motion parameters after 4 h of incubation at 38 °C were lower in samples with ≥5 × 10<sup>6</sup> PMN/ml (P < 0.05). For the clinical validation, semen samples from 305 beef bulls were evaluated. Unsatisfactory breeders (n = 83) had more CD45-positive cells (P = 0.016) and positive LEDT results (P = 0.008) than satisfactory breeders (n = 222). With CD45 immunostaining as the gold standard, the hemacytometer count had the highest clinical sensitivity (64.3 %) but the lowest specificity (73.3 %). A higher specificity was obtained with the peroxidase test (95.1 %) or semen cytology (98.8 %). In conclusion, the presence of ≥5 × 10<sup>6</sup> PMN/ml was associated with decreased semen quality in beef bulls. The hemacytometer count was the most sensitive bull-side test. But due to the low specificity, positive hemacytometer counts should be confirmed with the identification of peroxidase-positive cells or morphological identification of leucocytes on semen cytology. The CD45 immunostaining is the gold standard for the diagnosis of leucospermia in bulls.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142157550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-03DOI: 10.1016/j.theriogenology.2024.08.028
{"title":"The effect of resveratrol on the developmental competence of feline oocytes vitrified at the metaphase II stage","authors":"","doi":"10.1016/j.theriogenology.2024.08.028","DOIUrl":"10.1016/j.theriogenology.2024.08.028","url":null,"abstract":"<div><p>The objective of this research was to assess the viability and developmental potential of feline oocytes following in vitro maturation (IVM), vitrification, and post-warming incubation with resveratrol. In the first experiment, warmed oocytes were incubated with 0.2 μM, 2 μM, or 20 μM resveratrol for 2 h. Oocytes treated with 0.2 μM resveratrol had the highest viability (68.89 %), as assessed by fluorescein diacetate and ethidium bromide staining, while higher concentrations were associated with diminished oocyte viability. In the second experiment, the warmed oocytes were inseminated following the 2-h incubation with the three concentrations of resveratrol. The presumptive zygotes were then maintained in culture and their development evaluated. The highest cleavage rate was observed when the oocytes had been incubated with 0.2 μM resveratrol (88.34 %), which was higher than for the control group (without resveratrol (75 %)). Moreover, this concentration of resveratrol also augmented the blastocyst formation rate. While the vitrification of oocytes often results in diminished developmental potential in the ensuing embryos, attributed to cryopreservation-induced injury, the utilization of low concentrations of resveratrol enhances the procedure's efficacy.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142232988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-09-02DOI: 10.1016/j.theriogenology.2024.09.001
{"title":"Heat shock on bovine embryos from day 2.5–3 selects the most competent for progression to the blastocyst stage","authors":"","doi":"10.1016/j.theriogenology.2024.09.001","DOIUrl":"10.1016/j.theriogenology.2024.09.001","url":null,"abstract":"<div><p>Heat shock can impair embryo formation, while growth factors, such as colony-stimulating factor 2 (CSF2), modulate embryonic development. This study evaluated the effect of heat shock between days 2.5 and 3, as well as the impact of CSF2 at day 5 on bovine embryos cultured in a serum-free <em>in vitro</em> medium. The focus was on blastocyst development, the number of blastomeres, DNA fragmentation (TUNEL-positive cells), and mitochondrial activity. Heat shock reduced the proportion of cleaved embryos that developed into blastocysts (<em>P</em> = 0.0603). The resultant blastocysts exhibited a reduced number and proportion of TUNEL-positive cells in the trophectoderm (<em>P</em> = 0.0270 and <em>P</em> = 0.0240, respectively) and in the entire embryo (<em>P</em> = 0.0029 and <em>P</em> = 0.0031, respectively). Additionally, mitochondrial activity was lower in blastocysts derived from heat-shocked embryos (<em>P</em> = 0.0150) and further reduced in embryos exposed to both heat shock and CSF2 (<em>P</em> = 0.0415). In conclusion, the exposure of cleaved embryos to heat shock reduced their development to the blastocyst stage. However, the resulting blastocysts showed decreased DNA fragmentation and mitochondrial activity.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142146296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TheriogenologyPub Date : 2024-08-31DOI: 10.1016/j.theriogenology.2024.07.028
{"title":"Impact of polyamine supplementation on GnRH expression, folliculogenesis, and puberty onset in young mice","authors":"","doi":"10.1016/j.theriogenology.2024.07.028","DOIUrl":"10.1016/j.theriogenology.2024.07.028","url":null,"abstract":"<div><h3>Background</h3><p>The hypothalamic-pituitary-gonadal (HPG) axis is pivotal in regulating reproductive functions, with gonadotropin-releasing hormone (GnRH) acting as a central regulator. Recently, polyamines have been shown to regulate the HPG axis, including GnRH expression and ovarian biology in old and adult rodents. The present study firstly highlights the age-specific variation in the polyamine and their corresponding biosynthetic enzymes in the ovary during aging, and further, the study focuses on the effect of polyamines, putrescine, and agmatine, in young female mice.</p></div><div><h3>Method and result</h3><p>Immunofluorescence analysis revealed age-related differences in the expression of ornithine decarboxylase 1 (ODC1), spermine (SPM), and spermidine (SPD) in the ovaries, with adult mice exhibiting significantly higher expression levels compared to young and old mice. Likewise, qPCR analysis showed the mRNA levels of <em>Odc1</em>, Spermidine synthase (<em>Srm</em>), and Spermine synthase (<em>Sms</em>) show a significant increase in adult ovaries, which is then followed by a significant decline in old age. Histological examination demonstrated morphological alterations in the ovaries with age, including decreased follicle numbers and increased stromal cells in old mice. Furthermore, treatment with putrescine, a polyamine, in young mice resulted in larger ovaries and increased follicle numbers compared to controls. Additionally, serum levels of gonadotropin-releasing hormone (GnRH) and progesterone (P4) were measured, showing elevated levels in polyamine-treated mice. GnRH mRNA expression also increased significantly. Gene expression analysis revealed upregulation of genes associated with folliculogenesis such as <em>Fshr, Bmp15, Gdf9, Amh, Star, Hsdb3,</em> and <em>Plaur</em> in the ovaries and onset of puberty such as <em>Tac2</em>, and <em>Kiss1</em>, and a decrease in <em>Mkrn3</em> in the hypothalamus of polyamine-treated mice.</p></div><div><h3>Conclusion</h3><p>This study investigates the effect of polyamines in young immature female mice, shedding light on their role in upregulating GnRH, and enhancing folliculogenesis. Overall, these findings suggest that polyamines play a crucial role in ovarian aging and HPG axis regulation, offering potential therapeutics to reinstate fertility in reproductively challenged individuals.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142098193","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}