Effect of porcine spermatogonia germ cells on de novo morphogenesis of testis trough xenotransplatation.
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PROVIDER: PRJNA1197989 | ENA |
REPOSITORIES: ENA
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Project description:This study explores the role of porcine spermatogonia germ cells and testicular fibroblast in testis development and spermatogenesis through xenotransplantation. Previous research demonstrated the importance of Sertoli cells in maintaining testicular structure and function during testis development. However, the contribution of spermatogonia to these processes remains unclear. H&E staining revealed that FO tissues, no testicular structures were formed; instead, heterogeneous tissues with simple tubular formations were observed. Conversely, in GF tissues, structures resembling testicular tissue developed, displaying various stages of testis development including elongated spermatid. Immunofluorescence staining showed seminiferous tubules expressing PGP 9.5, VASA, and ACR2, confirming the presence of spermatogenesis. Interestingly, FO tissues were dominantly expressed ESR1, indicating bipotent gonadal property. RNA-seq analysis further revealed that GF tissue showed spermatogenesis-linked GOBPs with gene expression of mature Sertoli cells, including NR5A1, AR, and DMRT1. However, FO tissues displayed early tissue formation processes with expression of BMP4, WNT5A, ESR1, TGFBR3 and ACVR, generally inhibited in normal testis. These findings highlight the critical role of germ cells in formation of testicular structures through xenotransplantation and suggest the potential for spermatogenesis within these structures. However, further research is needed to verify the integrity and fertilizing capacity of the sperm, as these aspects have not been confirmed in this study.
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Project description:To identify the transcripts preferentially expressed in type A spermatogonia in rainbow trout immature testis, we compared the transcriptome between type A spermatogonia and testicular somatic cells by microarry analysis. We used fluorescence-activated cell sorting (FACS) to isolate type A spermatogonia or testicular somatic cells from the fishes carrying the transgene, pvasa-GFP, which expresses GFP in spermatogonia. RNA from type A spermatogonia and from testicular somatic cells were hybridized to a microarray after Cy3 labeling.
2012-06-11 | E-GEOD-38647 | biostudies-arrayexpress
Project description:To identify the transcripts preferentially expressed in type A spermatogonia in rainbow trout immature testis, we compared the transcriptome between type A spermatogonia and testicular somatic cells by microarry analysis. We used fluorescence-activated cell sorting (FACS) to isolate type A spermatogonia or testicular somatic cells from the fishes carrying the transgene, pvasa-GFP, which expresses GFP in spermatogonia.
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Project description:Spermatogenesis is a complex sperm-generating process involving the mitosis of spermatogonia, meiosis of spermatocytes and spermiogenesis of spermatids. Elucidating the phosphorylation-based regulations should advance our understanding of the underlying molecular mechanisms. Here we present an integrative study of phosphorylation events in the testis. Large-scale phosphoproteome profiling in the adult mouse testis identified 17,829 phosphorylation sites in 3,955 phosphoproteins.
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Project description:In adult mouse testis, undifferentiated spermatogonia include GFRα1+ and NGN3+ populations. The former mainly act as self-renewing stem cells that support steady-state spermatogenesis. Whilst, the latter are primed for differentiation, but retain the potential of self-renewal and contribute to regeneration and post-transplantation colony formation (Nakagawa et al. science .2010; Hara et al. cell stem cell. 2014). And, NGN3+ cells can differentiate to KIT+ cells, which are also mentioned differentiated spermatogonia, by retinoic acid signal. The gene expression profiles of GFRα1+, NGN3+ and KIT+ spermatogonia, as well as the whole testis samples, were investigated here by microarray (Ikami et al. development. 2015).
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Project description:To reveal distinct transcriptomes associated with spermatogonial stem cell renewal vs. initiation of differentiation, single-cell transcriptomes from P6 ID4-EGFP+ spermatogonia (sorted for brightest or dimmest) or unselected testis cells were used for Drop-Seq analysis. The GFP-bright and dim phenotypes exhibit distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the 10x Genomics Chromium (a commercial Drop-Seq variant) to perform single-cell RNA-seq
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Project description:Full title: Prepubertal Human Spermatogonia and Mouse Gonocytes Share Conserved Gene Expression of Germline Stem Cell Regulatory Molecules Oligonucleotide Microarray Analysis Reveals Novel Genes Expressed in Prepubertal Human Spermatogonia and Mouse Gonocytes Four hundred cells were selected for each group of germ cells or testis somatic cells from three independent testis cell preparations from both human (age 2, 8 and 10 yr) and mouse (age all 3 days) Selected germ and somatic cells were placed in 25 µl Trizol reagent (Invitrogen) immediately after isolation Oligonucleotide microarray was utilized to identify genes enriched in human and mouse selected germ cell populations and testis somatic cells.
2009-11-06 | E-GEOD-18914 | biostudies-arrayexpress
Project description:Embryonic day 13 (E13), E14, and E16 rat testes and ovaries were used for microarray analysis, as well as E13 testis organ cultures that undergo testis morphogenesis and develop seminiferous cords in vitro. A list of 109 genes resulted from a selective analysis for genes present in male gonadal development and with a 1.5-fold change in expression between E13 and E16. Characterization of these 109 genes potentially important for testis development revealed that cytoskeletal-associated proteins, extracellular matrix factors, and signaling factors were highly represented. Throughout the developmental period (E13-E16), sex-enriched transcripts were more prevalent in the male with 34 of the 109 genes having testis-enriched expression during sex determination. In ovaries, the total number of transcripts with a 1.5-fold change in expression between E13 and E16 was similar to the testis, but none of those genes were both ovary enriched and regulated during the developmental period. Genes conserved in sex determination were identified by comparing changing transcripts in the rat analysis herein, to transcripts altered in previously published mouse studies of gonadal sex determination. A comparison of changing mouse and rat transcripts identified 43 genes with species conservation in sex determination and testis development. Profiles of gene expression during E13-E16 rat testis and ovary development are presented and candidate genes for involvement in sex determination and testis differentiation are identified. Analysis of cellular pathways did not reveal any specific pathways involving multiple candidate genes. However, the genes and gene network identified influence numerous cellular processes with cellular differentiation, proliferation, focal contact, RNA localization, and development being predominant. Keywords: expression analysis, testis, ovary, sex determination
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Project description:Human adult spermatogenesis involves a balance of spermatogonial stem cell self renewal and differentiation, alongside complex germline-niche interactions. To better understand, we performed single cell RNA sequencing of ~7000 testis cells from three healthy men of peak reproductive age. Our analyses revealed multiple distinctive transcriptional ‘states’ of self-renewing and differentiating spermatogonia, the cellular stages of gametogenesis, five niche cells (Leydig, Myoid, Sertoli, Endothelial, macrophage) and insights into germline-niche communication. Spermatogenesis was reconstructed computationally, which identified sequential coding, noncoding, and repeat-element transcriptional signatures. A new, developmentally early and likely quiescent spermatogonial state is identified (GFRA1-/ETV5-/ID4+/UTF1+/FGFR3+). Notably, certain epigenetic features combined with nascent transcription analyses suggest considerable plasticity within certain spermatogonial populations/states. Key findings were validated via RNA and protein staining. Taken together, we provided the first “Cell Atlas” of the adult human testis, and provide multiple new insights into germ cell development and germ cell – niche interaction.
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