ABSTRACT: Tobacco-induced microRNAs profile alterations in human spermatozoa: a preliminary study for further knowledge of toxical spermatogenesis impairment
Project description:Tobacco-induced microRNAs and mRNAs profile alterations in human spermatozoa: a preliminary study for further knowledge of toxical spermatogenesis impairment
Project description:Tobacco-induced mRNAs profile alterations in human spermatozoa: a preliminary study for further knowledge of toxical spermatogenesis impairment
Project description:Tobacco smoking generates deleterious effects on human semen quality but mechanisms by which cigarettes smoking can impact spermatogenesis are poorly understood. Recent works have shown that spermatozoa RNAs can be used to understand mechanisms involved in tobacco induced spermatogenesis impairment. We performed a prospective study of 8 smoker and 8 non-smoker patients in an university hospital. All patients were selected according to an occupational exposure standardized questionnaire and the sperm parameters. We performed gene expression and miRNA microarrays using RNA extracts from spermatozoa of 8 smokers and 8 non-smokers. Quantification of selected miRNA was performed using quantitative RT-PCR. We show that 16 genes were differentially expressed between smokers and non-smokers, of which 5 were upregulated and 11 were down regulated in smokers. 23 microRNAs were differentially expressed, of which 16 were upregulated and 7 were down regulated in smokers. Quantitative RT-PCR confirmed the down regulation in smokers for 3 microRNAs. Moreover in smokers, one of the upregulated genes is a putative target for one down regulated microRNA. This is a preliminary and innovating study on spermatozoa RNA extracts in the field of infertility. This preliminary study shows that large scale approaches are non invasive diagnostic tools that may help elucidate the mechanisms that mediate the response to tobacco smoke exposure on human spermatogenesis. This could be used to design biomarkers of human spermatogenetic damage, and help elucidate the fine regulatory mechanisms that mediate responses to environmental agent exposure during human spermatogenesis.
Project description:Tobacco smoking generates deleterious effects on human semen quality but mechanisms by which cigarettes smoking can impact spermatogenesis are poorly understood. Recent works have shown that spermatozoa RNAs can be used to understand mechanisms involved in tobacco induced spermatogenesis impairment. We performed a prospective study of 8 smoker and 8 non-smoker patients in an university hospital. All patients were selected according to an occupational exposure standardized questionnaire and the sperm parameters. We performed gene expression and miRNA microarrays using RNA extracts from spermatozoa of 8 smokers and 8 non-smokers. Quantification of selected miRNA was performed using quantitative RT-PCR. We show that 16 genes were differentially expressed between smokers and non-smokers, of which 5 were upregulated and 11 were down regulated in smokers. 23 microRNAs were differentially expressed, of which 16 were upregulated and 7 were down regulated in smokers. Quantitative RT-PCR confirmed the down regulation in smokers for 3 microRNAs. Moreover in smokers, one of the upregulated genes is a putative target for one down regulated microRNA. This is a preliminary and innovating study on spermatozoa RNA extracts in the field of infertility. This preliminary study shows that large scale approaches are non invasive diagnostic tools that may help elucidate the mechanisms that mediate the response to tobacco smoke exposure on human spermatogenesis. This could be used to design biomarkers of human spermatogenetic damage, and help elucidate the fine regulatory mechanisms that mediate responses to environmental agent exposure during human spermatogenesis.
Project description:MicroRNAs (miRNAs) are involved in nearly every biological process examined to date. Mounting evidence show that some spermatozoa specific miRNAs play important roles in the regulation of spermatogenesis and germ cells development, but little is known of the exact identity and function of miRNA in sperm cells or their potential involvement in spermatogenesis and germ cells development. Here, we investigated the spermatozoa miRNA profiles using illumina deep sequencing combined with bioinformatic analysis using zebrafish as a model system. Deep sequencing of small RNAs yielded 12 million raw reads from zebrafish spermatozoa. Analysis showed that the noncoding RNA of the spermatozoa included tRNA, rRNA, snRNA, snoRNA and miRNA. By mapping to the zebrafish genome, we identified 400 novel and 204 conserved miRNAs which could be grouped into 104 families, including zebrafish specific families, such as mir-731, mir-724, mir-725, mir-729 and mir-2185. We report the first characterization of the miRNAs profiling in zebrafish spermatozoa. The obtained spermatozoa miRNAs profiling will serve as valuable resources to systematically study spermatogenesis in fish and vertebrate. Examination of small RNA populations in zebrafish spermatozoa
Project description:MicroRNAs (miRNAs) are involved in nearly every biological process examined to date. Mounting evidence show that some spermatozoa specific miRNAs play important roles in the regulation of spermatogenesis and germ cells development, but little is known of the exact identity and function of miRNA in sperm cells or their potential involvement in spermatogenesis and germ cells development. Here, we investigated the spermatozoa miRNA profiles using illumina deep sequencing combined with bioinformatic analysis using zebrafish as a model system. Deep sequencing of small RNAs yielded 12 million raw reads from zebrafish spermatozoa. Analysis showed that the noncoding RNA of the spermatozoa included tRNA, rRNA, snRNA, snoRNA and miRNA. By mapping to the zebrafish genome, we identified 400 novel and 204 conserved miRNAs which could be grouped into 104 families, including zebrafish specific families, such as mir-731, mir-724, mir-725, mir-729 and mir-2185. We report the first characterization of the miRNAs profiling in zebrafish spermatozoa. The obtained spermatozoa miRNAs profiling will serve as valuable resources to systematically study spermatogenesis in fish and vertebrate.
Project description:In the present study, we evaluated the alteration of protein profile of spermatozoa during the different stages of cryopreservation i.e., freshly ejaculated sperm, equilibrated sperm, and cryopreserved sperm in crossbred bulls (Bos taurus * Bos indicus). It was found that the equilibration step of cryopreservation itself caused major changes in the proteome of spermatozoa. Therefore, cryopreservation protocols should be tailored in such a way that it minimize sperm proteome alterations.
Project description:Spermatogenesis is a highly complex biological process through which male sperm cells, or spermatozoa, are produced in the testes. Besides enabling the flow of genetic information, spermatogenesis also creates a potential template for inter- and transgenerational inheritance of gene-regulatory states. While extensively studied in mammals, current knowledge of anamniote spermatogenesis remains limited. Here we provide a comprehensive resource consisting of single-cell gene expression and chromatin accessibility data complemented by base-resolution DNA methylome profiling of sorted germ cell populations, obtained from zebrafish (Danio rerio) testes. We identify major germ and somatic cell types implicated in zebrafish spermatogenesis as well as transcriptional drivers associated with each cell type. Moreover, we describe a localised DNA methylome reconfiguration event associated with the spermatocyte stage, and both local and global changes in chromatin accessibility leading to chromatin compaction in spermatids. Finally, we identify loci that escape global chromatin compaction, and which remain accessible thus forming a potential template for the intergenerational transmission of epigenetic states. In summary, our high-resolution cellular atlas of zebrafish spermatogenesis constitutes a major community resource that will enable further studies of germ cell development, evolution, and that will deepen our understanding of diverse molecular mechanisms implicated in genetic and epigenetic inheritance.
Project description:Spermatogenesis is a highly complex biological process through which male sperm cells, or spermatozoa, are produced in the testes. Besides enabling the flow of genetic information, spermatogenesis also creates a potential template for inter- and transgenerational inheritance of gene-regulatory states. While extensively studied in mammals, current knowledge of anamniote spermatogenesis remains limited. Here we provide a comprehensive resource consisting of single-cell gene expression and chromatin accessibility data complemented by base-resolution DNA methylome profiling of sorted germ cell populations, obtained from zebrafish (Danio rerio) testes. We identify major germ and somatic cell types implicated in zebrafish spermatogenesis as well as transcriptional drivers associated with each cell type. Moreover, we describe a localised DNA methylome reconfiguration event associated with the spermatocyte stage, and both local and global changes in chromatin accessibility leading to chromatin compaction in spermatids. Finally, we identify loci that escape global chromatin compaction, and which remain accessible thus forming a potential template for the intergenerational transmission of epigenetic states. In summary, our high-resolution cellular atlas of zebrafish spermatogenesis constitutes a major community resource that will enable further studies of germ cell development, evolution, and that will deepen our understanding of diverse molecular mechanisms implicated in genetic and epigenetic inheritance.
Project description:Malnad Gidda is one among the 43 registered cattle breeds of India with unique traits and spread over Western Ghats and coastal regions of Karnataka state in India. Selection of highly elite fertile bulls for the breeding purpose is a critical control point in animal breeding programmes. Therefore, to characterize the semen proteome and to understand the semen biology of this breed, a comprehensive proteomic analysis of spermatozoa and seminal plasma has been carried out by employing SCX and bRPLC fractionation strategies in a mass-spectrometry platform. The semen samples from three Malnad Gidda bulls maintained at Southern Regional Station of ICAR-NDRI under standard managemental conditions were used in the study. The proteomic characterisation of semen from Malnad Gidda breed resulted in the identification of 5, 84,520 PSMs, from 24,467 peptides from 2,815 proteins in spermatozoa and identification of 2, 77,583 PSMs from 12,047 peptides, which resulted in 1,974 proteins from seminal plasma. Out of 2,815 proteins in spermatozoa and 1,974 proteins from seminal plasma, 969 proteins were common to both seminal plasma and spermatozoa. The biological processes and cellular localization of spermatozoa proteins were studied using DAVID tool and were further enriched the identical GO terms using REVIGO online tool. The functional enrichment analysis of identified proteins indicated their roles in the biological processes like sperm motility, spermatid development, spermatogenesis, and so on. GO studies showed the commonalities and differences in the molecular functions of the proteins exclusively identified in spermatozoa, seminal plasma and common proteins. This is the first proteomic investigation conducted on the semen samples of an Indian indigenous breed; therefore, we believe that our preliminary data should significantly advance our understanding of semen proteome of Indian cattle.