Project description:The aim of this study was to analyze the gene expression profile for three main cell lines (supporting, interstitial/stromal, and germ cells) isolated from developing gonads during the process of the testis cord and ovarian cyst formation (between TS13 and TS34).
Project description:The aim of this study was to analyze the gene expression profile for three main cell lines (supporting, interstitial/stromal, and germ cells) isolated from developing gonads at the critical period of sexual differentiation (between 11.5, and 13.5 dpc). Three cell lines (supporting, interstitial/stromal, and germ cells) were isolated from murine fetal XX and XY gonads at three time points (11.5, 12.5, and 13.5 dpc). Transgenic mouse strains with the expression of cell type specific fluorescent markers were used to isolate the cell lines. Cells were sorted using FACS method and then the RNA was extracted.
Project description:The goal of this study is to determine the complete gene expression profile for each cell type of the developing gonad during the critical window in which it adopts the testis or ovarian fate. Transgenic mice with cell type specific fluorescent markers were used to isolate germ cells, supporting cells, interstitial cells (including steroidogenic precursors), and endothelial cells in the developing testis and ovary. The gonads were dissociated in trypsin, and the fluorescent cells were isolated by FACS. The RNA was collected from the isolated cells and their gene expression profiles were determined by microarray analysis.
Project description:The aim of this study was to analyze the gene expression profile for three main cell lines (supporting, interstitial/stromal, and germ cells) isolated from developing gonads at the critical period of sexual differentiation (between 11.5, and 13.5 dpc).
Project description:NSAIDs and ACE that affect prostaglandin synthesis are widely used by pregnant women. Epidemiological studies have hypothesized a potential relation of testis dysgenesis syndromes such as cryptorchidism and hypospadias to exposure to these molecules during both the first and the second trimesters of gestation. To decipher whether the embryonic gonads themselves are targets for these molecules, we analysed the impact of precocious in utero exposure to NSAIDs and ACE alone or in combination on the early development of the testis during sex determination, using therapeutic doses similar to those administrated in human medications. We found that in utero exposure to ACE, aspirin or ibuprofen affects the germ cell proliferation in embryonic testis. The whole transcriptome of 13.5 dpc (days post coïtum) treated testis suggests different mechanisms of action of these drugs and a functional interaction between both molecules used in combination, in accelerating the germ cell differentiation. We identified that ACE and ibuprofen exposure through the up-regulation of Dnmt3L expression induces advanced epigenetic reprograming of the germline and enhanced glycogen storage within the testis cords through the activation of extracellular matrix genes expression. In addition, we identified for the first time the prostaglandin production pattern in the embryonic gonad and showed that PGD2, PGE2 and PGI2 were the targets of ACE and NSAIDs drugs. These features might affect the formation and maturation of postnatal testis and secondary reproductive organs leading to male infertility in adult age.
Project description:Human testis development in prenatal life involves complex changes in germline and somatic cell identity. To better understand, we profiled and analyzed ~32,500 single-cell transcriptomes of testicular cells from embryonic, fetal and infant stages. Our data shows that at 6-7 weeks post-fertilization as the testicular cords are established, the Sertoli and interstitial cells originate from a common heterogeneous progenitor pool, which then resolves into fetal Sertoli cells (expressing tube-forming genes) or interstitial cells (including Leydig-lineage cells expressing steroidogenesis genes). Almost ten weeks later, beginning at 15-16 weeks post-fertilization, the male primordial germ cells exit mitosis, downregulate pluripotent transcription factors and transition into cells that strongly resemble the ‘State 0’ spermatogonia originally defined in the infant and adult testes. Therefore, we termed these fetal spermatogonia ‘State f0’. Taken together, we reveal multiple insights into the coordinated and temporal development of the embryonic, fetal and postnatal male germline together with the somatic niche.
Project description:We collected whole genome testis expression data from hybrid zone mice. We integrated GWAS mapping of testis expression traits and low testis weight to gain insight into the genetic basis of hybrid male sterility.
Project description:Gonadal sex differentiation – testis versus ovary formation – is a fundamental process required for reproduction and evolution. Reflecting this importance, the embryonic gonads of vertebrate species comprise the same key cell types; germ cells, supporting cells and interstitial steroidogenic cells. Remarkably, the genetic triggers for gonadal sex differentiation vary across species (the SRY gene in mammals, DMRT1 in birds and some turtles, temperature in many reptiles, AMH and various other genes in fishes). Despite this variation, the cell biology of gonadal development was long thought to be largely conserved. Here, we present a comprehensive analysis of gonadal sex differentiation, using the chicken embryo as a model and considering the entire gonad. We sampled over 30,000 cells across several developmental stages, prior, during and after the onset of gonadal sex differentiation. The data provide several new insights into cell lineage specification during vertebrate gonadogenesis. Combining lineage tracing with single cell transcriptomics, the data show that somatic supporting cells of the embryonic chicken gonad do not derive from the coelomic epithelium, in contrast to other vertebrates studied. Instead, the early somatic precursors cells of the gonads in both sexes derive from a DMRT1+/PAX2+/WNT4+/OSR1+ mesenchymal cell population. In particular, PAX2 marks immigrating mesenchymal cells that give rise to the supporting cell lineage. We find a greater complexity of gonadal cell types than previously thought, including the identification of two distinct sub-populations of Sertoli cells in developing testes, and derivation of embryonic steroidogenic cells from a differentiated supporting cell lineage. We provide significantly improved resolution of gonadal cell types and identify several new gonadal marker genes. Altogether, these results indicate that, just as the genetic trigger for sex differs across vertebrate groups, cell lineage specification in the gonad may also vary substantially.
Project description:Gonadal soma-derived factor (gsdf) has been demonstrated as essential for testicular differentiation in medaka (Oryzias latipes). To understand the protein dynamics of Gsdf in spermatogenesis regulation, we used a His-tag “pull-down” assay coupled with shotgun LC-MS/MS to identify a group of potential interaction partners for Gsdf, which included cytoplasmic dynein light chain 2, eukaryotic polypeptide elongation factor 1 alpha (eEF1), and actin filaments in mature medaka testis. Alike the interaction with TGF-dynein critical for spermatogonial division in Drosophila melanogaster, the physical interaction of Gsdf & dynein and Gsdf & eEF1 were identified through a yeast 2-hybrid (Y2H) screening of an adult testis cDNA library using Gsdf as bait, and were verified by a paired Y2H assay. Co-immunoprecipitation of Gsdf and eEF1was defined in adult testes, as supporting the requirement of Gsdf and eEF1 interaction in testis development. Proteomics analysis and ultrastrutural observation showed that Gsdf deficiency activated eEF1-mediated protein synthesis and ribosome biogenesis, which in turn led to the differentiation of undifferentiated germ cell. Thus, our results provide a framework and new insight into the coordination of Gsdf (TGF and eEF1complex in the basic processes of germ cell proliferation, and transcriptional and translational control of sexual RNA, which may be fundamentally conserved across phyla during sex differentiation.