title: Expression profiling identifies Sertoli and Leydig cell genes as Fsh targets in adult zebrafish testis.
abstract: Spermatogonial stem cells are quiescent, undergo self-renewal or differentiating divisions, thereby forming the cellular basis of spermatogenesis. This cellular development is orchestrated by follicle-stimulating hormone (FSH), through the production of Sertoli cell-derived factors, and by Leydig cell-released androgens. Here, we investigate the transcriptional events induced by Fsh in a steroid-independent manner on the restart of zebrafish (Danio rerio) spermatogenesis ex vivo, using testis from adult males where type A spermatogonia were enriched by estrogen treatment in vivo. Under these conditions, RNA sequencing preferentially detected differentially expressed genes in somatic/Sertoli cells. Fsh-stimulated spermatogonial proliferation was accompanied by modulating several signaling systems (i.e. Tgf-β, Hedgehog, Wnt and Notch pathways). In silico protein-protein interaction analysis indicated a role for Hedgehog family members potentially integrating signals from different pathways during fish spermatogenesis. Moreover, Fsh had a marked impact on metabolic genes, such as lactate and fatty acid metabolism, or on Sertoli cell barrier components. Fish Leydig cells express the Fsh receptor and one of the most robust Fsh-responsive genes was insulin-like 3 (insl3), a Leydig cell-derived growth factor. Follow-up work showed that recombinant zebrafish Insl3 mediated pro-differentiation effects of Fsh on spermatogonia in an androgen-independent manner. Our experimental approach allowed focusing on testicular somatic genes in zebrafish and showed that the activity of signaling systems known to be relevant in stem cells was modulated by Fsh, providing promising leads for future work, as exemplified by the studies on Insl3.
authors: Reproductive Biology Group, Division of Developmental Biology, Department of Biology, Faculty of Science, Utrecht University, Utrecht, The Netherlands.
authors: Diego
authors: D
authors: Crespo
authors: Reproductive Biology Group, Division of Developmental Biology, Department of Biology, Faculty of Science, Utrecht University, Utrecht, The Netherlands.
authors: Luiz H C
authors: LHC
authors: Assis
authors: Research Group Reproduction and Developmental Biology, Institute of Marine Research, Bergen, Norway.
authors: Tomasz
authors: T
authors: Furmanek
authors: Reproductive Biology Group, Division of Developmental Biology, Department of Biology, Faculty of Science, Utrecht University, Utrecht, The Netherlands.
authors: Jan
authors: J
authors: Bogerd
authors: Reproductive Biology Group, Division of Developmental Biology, Department of Biology, Faculty of Science, Utrecht University, Utrecht, The Netherlands; Research Group Reproduction and Developmental Biology, Institute of Marine Research, Bergen, Norway. Electronic address: r.w.schulz@uu.nl.
authors: Rüdiger W
authors: RW
authors: Schulz
mesh_terms: D000375:Aging / Q000235:genetics; D000728:Androgens / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation / Q000235:genetics; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D057809:HEK293 Cells; D006801:Humans; D007985:Leydig Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D060066:Protein Interaction Maps; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D012708:Sertoli Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013091:Spermatogenesis / Q000187:drug effects / Q000235:genetics; D013093:Spermatogonia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013737:Testis / Q000187:drug effects / Q000378:metabolism*; D059467:Transcriptome / Q000235:genetics; D015027:Zebrafish / Q000235:genetics* / Q000254:growth & development*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000728:Androgens; D012333:RNA, Messenger; D011994:Recombinant Proteins; D029961:Zebrafish Proteins; D005640:Follicle Stimulating Hormone
keywords: Follicle-stimulating hormone; RNA sequencing; Spermatogenesis; Zebrafish