MicroRNA-202 is a gatekeeper of meiosis initiation and regulates recombination in male germ line
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE126936 | GEO | 2020/02/15
REPOSITORIES: GEO
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Project description:MicroRNA-202 is a gatekeeper of meiosis initiation and regulates recombination in male germ line
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Project description:KDM2A/FBXL11 is a Jumonji-domain containing lysine demethylase catalyzing the removal of mono- and di-methyl modifications of histone H3 lysine 36. While Kdm2a is required for mouse embryogenesis, its role in adult physiology is unknown. Using conditional deletion approaches, we demonstrate that Kdm2a deficiency causes testicular atrophy and male infertility. Though spermatogonial stem cells were unaffected, proliferating and differentiating spermatogonia suffered from delayed cell cycle progression and apoptosis, which correlated with upregulated expression of several cell cycle inhibitors. Loss of Kdm2a in spermatocytes disrupted progression through meiotic prophase, as shown by impaired chromosome synapsis and processing of meiotic double strand breaks, and by altered chromatin states. Correspondingly, Kdm2a mutant spermatocytes failed to activate numerous genes controlling these processes. RNA-sequencing analyses on purified spermatogonia and spermatocytes showed that during normal spermatogonial differentiation over 700 genes undergo repression, which is controlled by KDM2A. CpG-rich promoter genes upregulated in Kdm2a deficient cells are marked by Polycomb Repressive Complexes (PRC) and associated modifications in wildtype male germ cells, suggesting that KDM2A is required for PRC-mediated repression. Our study thus identifies critical roles for KDM2A in coordinating gene expression programs during spermatogonial differentiation and meiosis, which are essential for male germ cell development.
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Project description:KDM2A/FBXL11 is a Jumonji-domain containing lysine demethylase catalyzing the removal of mono- and di-methyl modifications of histone H3 lysine 36. While Kdm2a is required for mouse embryogenesis, its role in adult physiology is unknown. Using conditional deletion approaches, we demonstrate that Kdm2a deficiency causes testicular atrophy and male infertility. Though spermatogonial stem cells were unaffected, proliferating and differentiating spermatogonia suffered from delayed cell cycle progression and apoptosis, which correlated with upregulated expression of several cell cycle inhibitors. Loss of Kdm2a in spermatocytes disrupted progression through meiotic prophase, as shown by impaired chromosome synapsis and processing of meiotic double strand breaks, and by altered chromatin states. Correspondingly, Kdm2a mutant spermatocytes failed to activate numerous genes controlling these processes. RNA-sequencing analyses on purified spermatogonia and spermatocytes showed that during normal spermatogonial differentiation over 700 genes undergo repression, which is controlled by KDM2A. CpG-rich promoter genes upregulated in Kdm2a deficient cells are marked by Polycomb Repressive Complexes (PRC) and associated modifications in wildtype male germ cells, suggesting that KDM2A is required for PRC-mediated repression. Our study thus identifies critical roles for KDM2A in coordinating gene expression programs during spermatogonial differentiation and meiosis, which are essential for male germ cell development.
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Project description:Vertebrate spermatogonial stem cells maintain sperm production over the lifetime of an animal but fertility declines with age. While morphological studies have informed our understanding of typical spermatogenesis, the molecular and cellular mechanisms underlying the maintenance and decline of spermatogenesis are not yet understood. We used single-cell RNA sequencing to generate a developmental atlas of the aging zebrafish testis. All testes contained spermatogonia, but we observed a progressive decline in spermatogenesis that correlates with age. Testes from some older males only contained spermatogonia and a reduced population of spermatocytes. Spermatogonia in older males are transcriptionally distinct from spermatogonia in testes capable of robust spermatogenesis. Immune cells including macrophages and lymphocytes drastically increase in abundance in testes that cannot complete spermatogenesis. Our developmental atlas reveals the cellular changes as the testis ages and defines a molecular roadmap for the regulation of spermatogenesis.
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Project description:Prophase I of male meiosis involves dynamic chromosome segregation processes during early spermatogenesis, including synapsis, meiotic recombination, and cohesion. Genetic defects in genes participating in these processes consistently cause reproduction failure in mice. To identify candidate genes responsible for infertility in humans, we performed expression profiling of mouse spermatogenic cells undergoing meiotic prophase I. Cell fractions enriched in spermatogonia, leptotene/zygotene spermatocytes, or pachytene spermatocytes were separately isolated from mouse testes for RNA extraction. To minimize the contamination of other cell types, we fractionated the testicular cells undergoing the first round of spermatogenesis using Percoll gradient procedure. The cell fractions were characterized by morphological analysis by phase contrast microscopy and Nomarski interference microscopy, and then expression of cell lineage- and spermatogenesis stage-specific genes were examined by RT-PCR. The most enriched fractions for spermatogonia (fraction2 from day8), leptotene/zygotene spermatocytes (fraction5 from day12), and pachytene spermatocytes (fraction8 from day15) were subjected to hybridization on Affymetrix microarrays.
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Project description:MicroRNA is essential for the process of spermatogonesis, however analysis of its change in expression within germ cells during this process has been limited. We set out to examine the change in the miRNA expression profile of highly enriched mouse germ cell populations in vairous developmental stages.These inlcuded postnatal gonocytes, day 7-9 spermatogonia as well as pachytene spermatocytes and round spermatids from adult mice Gonocytes (day1) and spermatogonia (day 7-9), pachytene spermatocytes and round spermadits (adult) were enrichged by 2-4% BSA gradient sedementation. Three biological replicates of each cell type were included.
2012-03-17 | E-GEOD-36566 | biostudies-arrayexpress
Project description:We report bulk RNA-sequencing data for undifferentiated and defined subpopulations of differentiating murine spermatogonia. We have sequenced two replicates each of wild-type and STRA8 knock-out sets of spermatogonia to assess transcriptional changes that occur throughout spermatogonial development prior to meiotic entry.
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Project description:Meiosis is a specialized form of cell division essential for sexual reproduction. The unique cellular events of meiosis have been extensively studied in model eukaryotes, including mammals. In mammals, the molecular and cellular changes that germ cells must undergo to transition from mitotic spermatogonia to meiotic spermatocytes remain largely undefined. Retinoic acid (RA) has been proposed as the meiosis inducing substance (MIS) required for female and male meiotic initiation. However, evidence for this presumption is based on studies of fetal gonads – meiosis is initiated in fetal ovaries but suppressed in fetal testes. Critically, the requirement for RA in meiosis has never been formally tested because germ cell differentiation and meiotic initiation in the fetal ovary are temporally inseparable. Using the postnatal mouse testis, where differentiation and meiotic initiation are temporally distinct (8.6 days apart), we test the hypothesis that RA is dispensable for initiation and progression through meiosis. Complementary in vivo experimental models were designed to address the hypothesis. In the first, “RA-sufficient” (RA-suf) model, spermatogenesis was synchronized by administration of potent and selective RA synthesis inhibitor; as a result, testes contained only undifferentiated spermatogonia. Then, the inhibitor was discontinued, and mice were given a single dose of exogenous RA to initiate spermatogonial differentiation. Germ cells completed differentiation, entered and completed meiosis, and formed haploid gametes at predicted timepoints. In the second, “RA-deficient” (RA-def) model, mice were treated as above, except the inhibitor was administered continuously. The lack of testicular RA in this model was confirmed in multiple ways, including assessment of RA-responsive gene expression, in vitro serum-free cultures, and dose-response experiments. To our surprise, germ cells in RA-def testes completed differentiation, underwent meiotic DNA replication, completed the landmark cytological processes of meiosis, and formed postmeiotic haploid spermatids, all on the predicted timeline of the RA-suf model. To define the effect of RA-deficiency on gene expression during meiotic initiation, large, highly enriched populations of premeiotic spermatogonia from RA-suf mice, and meiotic preleptotene spermatocytes from both models were FACS-isolated and subjected to bulk RNA-seq. Interestingly, analyses revealed few significant differences in expression of characteristic meiotic genes at the mRNA level. However, genes that were significantly different between RA-suf and RA-def preleptotene spermatocytes were associated with spermiogenesis functions. Taken together, our data reveal that, after initiating spermatogonial differentiation, RA is dispensable for initiation, progression through, and completion of meiosis in the male germline.
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