Effects of cryoprotectant treatments on bovine sperm function and osmolyte content.
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ABSTRACT:
SUBMITTER: Setyawan EE
PROVIDER: S-EPMC3735009 | biostudies-other | 2009 Sep
REPOSITORIES: biostudies-other
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Project description:Transient receptor potential channels-vanilloid receptor 1 (TRPV1) regulates thermotaxis in sperm-oriented motility. We investigated the role of membrane cholesterol (Chol) on TRPV1-mediated human sperm migration. Semen samples were obtained from five normozoospemic healthy volunteers. Sperm membrane Chol content, quantified by liquid chromatography-mass spectrometry, was modified by incubating cells with 2-hydroxypropyl-ß-cyclodextrin (CD) or the complex between CD and Chol (CD:Chol). The effect on sperm migration on a 10 μM capsaicin gradient (CPS), a TRPV1 agonist, was then investigated. Motility parameters were evaluated by Sperm Class Analyser. Intracellular calcium concentration and acrosome reaction were measured by staining with calcium orange and FITC-conjugated anti-CD46 antibody, respectively. TRPV1-Chol interaction was modelled by computational molecular-modelling (MM). CD and CD:Chol, respectively, reduced and increased membrane Chol content in a dose-dependent manner, resulting in a dose-dependent increase and reduction of sperm migration in a CPS gradient. MM confirmed a specific interaction of Chol with a TRPV1 domain that appeared precluded to the Chol epimer epicholesterol (Epi-Chol). Accordingly, CD:Epi-Chol was significantly less efficient than CD:Chol, in reducing sperm migration under CPS gradient. Chol inhibits TRPV1-mediated sperm function by directly interacting with a consensus sequence of the receptor.
| S-EPMC8003190 | biostudies-literature
Project description:Salpingitis is a common cause for subfertility and infertility both in humans and animals. However, the effects of salpingitis on tubal function and reproductive success are largely unknown. Therefore we set out to investigate the effects of inflammation on sperm and oocyte transport and gameto-maternal interaction in the oviduct using the bovine as a model. For this purpose, oviducts revealing mild (n?=?45), moderate (n?=?55) and severe (n?=?45) inflammation were obtained from cows immediately after slaughter and investigated by live cell imaging, histochemistry and scanning electron microscopy. Our studies showed that endometritis was always correlated with salpingitis. Moderate and severe inflammation caused a significant increase in the thickness of tubal folds (p?<?0.05). Severe inflammation was characterized by luminal accumulations of mucus and glycoproteins, increased apoptosis, loss of tight junctions and shedding of tubal epithelial cells. The mean ciliary beat frequency (CBF) in the ampulla was significantly reduced as compared to the controls (p?<?0.05). The higher the grade of inflammation, the lower was the CBF (p?<?0.001). In severe inflammation, spermatozoa were stuck in mucus resulting in decreased sperm motility. Our results imply that tubal inflammation impairs proper tubal function and leads to reduced sperm fertilizing capacity.
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Project description:BackgroundThe use of sex-sorted sperm in cattle assisted reproduction is constantly increasing. However, sperm fertility can substantially differ between unsorted (conventional) and sex-sorted semen batches of the same sire. Sperm microRNAs (miRNA) have been suggested as promising biomarkers of bull fertility the last years. In this study, we hypothesized that the miRNA profile of cryopreserved conventional sperm is related to bull fertility after artificial insemination with X-bearing sperm. For this purpose, we analyzed the miRNA profile of 18 conventional sperm samples obtained from nine high- (HF) and nine low-fertility (LF) bulls that were contemporaneously used to produce conventional and sex-sorted semen batches. The annual 56-day non-return rate for each semen type (NRRconv and NRRss, respectively) was recorded for each bull.ResultsIn total, 85 miRNAs were detected. MiR-34b-3p and miR-100-5p were the two most highly expressed miRNAs with their relative abundance reaching 30% in total. MiR-10a-5p and miR-9-5p were differentially expressed in LF and HF samples (false discovery rate < 10%). The expression levels of miR-9-5p, miR-34c, miR-423-5p, miR-449a, miR-5193-5p, miR-1246, miR-2483-5p, miR-92a, miR-21-5p were significantly correlated to NRRss but not to NRRconv. Based on robust regression analysis, miR-34c, miR-7859 and miR-342 showed the highest contribution to the prediction of NRRss.ConclusionsA set of miRNAs detected in conventionally produced semen batches were linked to the fertilizing potential of bovine sperm after sex-sorting. These miRNAs should be further evaluated as potential biomarkers of a sire's suitability for the production of sex-sorted sperm.
| S-EPMC7792310 | biostudies-literature
Project description:BACKGROUND: In previous studies using candidate gene approaches, low sperm count (oligospermia) has been associated with altered sperm mRNA content and DNA methylation in both imprinted and non-imprinted genes. We performed a genome-wide analysis of sperm DNA methylation and mRNA content to test for associations with sperm function. METHODS AND RESULTS: Sperm DNA and mRNA were isolated from 21 men with a range of semen parameters presenting to a tertiary male reproductive health clinic. DNA methylation was measured with the Illumina Infinium array at 27,000 CpG loci. Unsupervised clustering of methylation data differentiated the 21 sperm samples by their motility values. Recursively partitioned mixture modeling (RPMM) of methylation data resulted in four distinct methylation profiles that were significantly associated with sperm motility (P=0.01). Linear models of microarray analysis (LIMMA) was performed based on motility and identified 9,189 CpG loci with significantly altered methylation (Q<0.05) in the low motility samples, with many loci located in genes associated with subfertility and epigenetic regulation. In the low motility samples, the majority of disrupted CpG loci (80%) were hypomethylated. Of the aberrantly methylated CpGs, 194 were associated with imprinted genes almost equally distributed into hypermethylated (predominantly paternally expressed) and hypomethylated (predominantly maternally expressed) groups. Sperm mRNA was measured with the Human Gene 1.0 ST Affymetrix GeneChip Array. LIMMA analysis based on motility identified 20 candidate transcripts as differentially expressed in low motility sperm, including HDAC1 (NCBI 3065), SIRT3 (NCBI 23410), and DNMT3A (NCBI 1788). Altered expression of these epigenetic regulatory genes was associated with RPMM DNA methylation class. CONCLUSIONS: Using integrative genome-wide approaches to study epigenetic and gene expression patterns in human sperm we identified CpG methylation profiles and mRNA alterations associated with low sperm motility, and that low motility sperm may have aberrant genome-wide hypomethylation due to excess HDAC1 activity. See "summary" above
2011-06-02 | E-GEOD-26881 | biostudies-arrayexpress
Project description:BACKGROUND: In previous studies using candidate gene approaches, low sperm count (oligospermia) has been associated with altered sperm mRNA content and DNA methylation in both imprinted and non-imprinted genes. We performed a genome-wide analysis of sperm DNA methylation and mRNA content to test for associations with sperm function. METHODS AND RESULTS: Sperm DNA and mRNA were isolated from 21 men with a range of semen parameters presenting to a tertiary male reproductive health clinic. DNA methylation was measured with the Illumina Infinium array at 27,000 CpG loci. Unsupervised clustering of methylation data differentiated the 21 sperm samples by their motility values. Recursively partitioned mixture modeling (RPMM) of methylation data resulted in four distinct methylation profiles that were significantly associated with sperm motility (P=0.01). Linear models of microarray analysis (LIMMA) was performed based on motility and identified 9,189 CpG loci with significantly altered methylation (Q<0.05) in the low motility samples, with many loci located in genes associated with subfertility and epigenetic regulation. In the low motility samples, the majority of disrupted CpG loci (80%) were hypomethylated. Of the aberrantly methylated CpGs, 194 were associated with imprinted genes almost equally distributed into hypermethylated (predominantly paternally expressed) and hypomethylated (predominantly maternally expressed) groups. Sperm mRNA was measured with the Human Gene 1.0 ST Affymetrix GeneChip Array. LIMMA analysis based on motility identified 20 candidate transcripts as differentially expressed in low motility sperm, including HDAC1 (NCBI 3065), SIRT3 (NCBI 23410), and DNMT3A (NCBI 1788). Altered expression of these epigenetic regulatory genes was associated with RPMM DNA methylation class. CONCLUSIONS: Using integrative genome-wide approaches to study epigenetic and gene expression patterns in human sperm we identified CpG methylation profiles and mRNA alterations associated with low sperm motility, and that low motility sperm may have aberrant genome-wide hypomethylation due to excess HDAC1 activity.
2011-06-02 | GSE26881 | GEO
Project description:The phenomenon of aging arises from multiple, complex interactions causing dysfunction in cells and organs. In particular, fertility drastically decreases with age. Previously, we have demonstrated that the functional characteristics of the bovine oviduct and uterus change with the age-dependent upregulation of inflammation and noted that S100A9 triggers inflammatory responses in oviduct epithelial cells. In the present study, we investigated the hypothesis that S100A9 affects reproductive events to aspect such as sperm function, fertilization, and the development of the embryo in cows. To investigate the effect of S100A9 on bovine sperm, we incubated sperms in vitro with S100A9 for 5 h and observed significantly decreased sperm motility and viability. During in vitro fertilization, S100A9 treatment for 5 h did not affect the rate of fertilization, time of first division of embryos, or embryo development to blastocyst stage. Treatment of 2-cell stage embryos with S100A9 for 5 h significantly reduced the proportion of cells undergoing normal division (4-8 cell embryos) and embryo development to the blastocyst stage. In experiment involving 24 h treatment of 2-cell embryos, the development of all embryos stopped at the 2-cell stage in the S100A9-treated group. In blastocyst-stage embryos, S100A9 treatment significantly stimulated the expression of endoplasmic reticulum (ER) and the mRNA expression of ER stress markers, and activated caspase-3 with subsequent nuclear fragmentation. Pre-treatment with an ER stress inhibitor significantly suppressed caspase-3 activation by the S100A9 treatment, suggesting that S100A9 induces blastocyst dysfunction by apoptosis (via caspase-3 activation) depending on ER stress. These results indicate that direct exposure to S100A9 exerted adverse effects on sperm function and embryo development. These findings suggest that excessive dose of S100A9 may have an adverse effect to the reproductive machinery by inducing inflammation and tissue dysfunction.
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Project description:In artificial insemination the use of sex-sorted bovine sperm results in reduced conception, the causes of which are only partly understood. Therefore, we set out to investigate the effects of sexing on bovine sperm function and early embryonic development. Computer-assisted semen analysis (CASA) of sperm of the same bulls (n = 5), before and after sexing, demonstrated significantly reduced fast (A) and slow (B) progressively motile sperm (p < 0.05) after sexing. Sexed-sperm also revealed significantly less hyperactivated sperm (p < 0.05). As shown by time-lapse videomicroscopy of in vitro produced embryos (n = 360), embryos derived from sexed-sperm displayed significantly increased incidences of arrest at the 4-cell stage (p < 0.05). The relative risk for shrinkage/fusion of blastomeres with subsequent lysis was 1.71 times higher in the embryos derived from sexed-sperm as compared to conventional embryos (p < 0.05) resulting in significantly reduced blastocyst rates (p < 0.001). The relative risk for cleavage was 2.36 times lower in the embryos derived from sex-sorted sperm (p < 0.001). Additionally, sexed-sperm-derived embryos showed reduced survival times (hazard ratio HR = 1.54, p < 0.001) which were bull dependent (p < 0.001). However, the percentage of apoptotic cells was similar to conventional embryos. Furthermore, embryos derived from sexed-sperm were found to reach developmental stages at similar timings as conventional embryos. Our results suggest that reduced conception rates after sexing are due to altered sperm morphokinetics, decreasing the chance of sperm to reach and fertilise the oocyte, and aberrant early embryonic development.
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Project description:Mammalian sperm motility is getting more relevant due to rising infertility rates worldwide, generating the need to improve conventional analysis and diagnostic approaches. Nowadays, computer assisted sperm analysis (CASA) technologies represent a popular alternative to manual examination which is generally performed by observing sperm motility in very confined geometries. However, under physiological conditions, sperm describe three-dimensional motility patterns which are not well reconstructed by the limited depth of standard acquisition chambers. Therefore, affordable and more versatile alternatives are needed. Here, a motility analysis in unconfined conditions is proposed. In details, the analysis is characterized by a significant longer duration -with respect to conventional systems- with the aim to observe eventually altered motility patterns. Brightfield acquisition in rectangular glass capillaries captured frozen-thawed bovine spermatozoa which were analyzed by means of a self-written tracking routine and classified in sub-populations, based on their curvilinear velocity. To test the versatility of our approach, cypermethrin -a commonly used pesticides- known to be responsible for changes in sperm motility was employed, assessing its effect at three different time-steps. Experimental results showed that such drug induces an increase in sperm velocity and progressiveness as well as circular pattern formation, likely independent of wall interactions. Moreover, this resulted in a redistribution of sperm with the rapid class declining in number with time, but still showing an overall velocity increase. The flexibility of the approach permits parameter modifications with the experimental needs, allowing us to conduct a comprehensive examination of sperm motility. This adaptability facilitated data acquisition which can be computed at different frame rates, extended time periods, and within deeper observation chambers. The suggested approach for sperm analysis exhibits potential as a valuable augmentation to current diagnostic instruments.
| S-EPMC11019440 | biostudies-literature
Project description:Purpose: to construct a library of miRNAs in bovine sperm was constructed using Illumina high‐throughput sequencing technology. Result: Unique sequences that were 18–26 nucleotides in length were mapped to specific precursors in miRBase 20.0 using BLAST. A total of 951 known miRNAs and 8 novel, highly expressed miRNA candidates were identified.
2021-02-26 | GSE167556 | GEO
Project description:Sperm performance is often tightly linked to male reproductive success. In many demersal gobiid fishes, the male attaches sperm embedded in a mucus produced by sperm-duct glands to the nest substrate before spawning takes place. Sperm are activated as the mucus and embedded gland content dissolve into the water. To test the importance of gland content on sperm function in Pomatoschistus minutus, a marine fish with external fertilization, we used a paired experimental design, with spermatozoa tested with and without sperm-duct gland content mixed into seawater. We measured sperm velocity, percentage of motile sperm and sperm viability over time. Sperm were found to swim 7.3% faster when gland content was mixed in the seawater. Percentage motile sperm was unaffected by the gland content. Sperm viability in seawater exceeded 24?h, but was unaffected by the gland content. An increase in sperm velocity of similar magnitude as found here has been shown by others to increase fertilization success. Since velocity-boosting properties of sperm-duct gland content have now been found in three distantly related goby species, this trait appears to be conserved across the Gobiidae family and may aid in reproduction across a range of species and environments.This article has an associated First Person interview with the first author of the paper.
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