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Title

Molecular characterization and expression of nanos2 AND gfra1a during testicular development of theh European Seabass (Dicentrarchus labrax L.)

AuthorsPrat, F.; Simón-Díaz, M.; Martos-Sitcha, Juan Antonio ; Gómez, Ana
Issue DateSep-2019
CitationXII Conference of the Iberian Association for Comparative Endocrinology (2019)
AbstractSpermatogenesis starts when undifferentiated type A spermatogonia (und-SpgA), also known as spermatogonial stem cells (SSC), become differentiated SpgA (diff-SpgA). Expression of nanos2 in und-SpgA has been observed in several species and it is recognized as a spermatogonial stem cell marker. In the mammalian testis, it is well established that Sertoli cells produce GDNF (glial cell line-derived neurotrophic factor) that binds the GFRA1 (GDNF family receptor alpha-1), which is expressed by und-SpgA promoting its self-renewal and maintenance. A few recent studies have addressed the potential involvement of the Gdnf-Gfra1 signaling pathway in the regulation of gametogenesis in cartilaginous and bony fishes. As a first step to determine nanos2 and gfra1a involvement in the self-renewal and maintenance of the und-SpgA preventing spermatogonial differentiation in the European seabass, the aim of this work was to characterize and study their expression at different stages of testicular development. By blast analysis against the NCBI whole-genome shotgun contigs (wgs) database for D. labrax (sequences CBXY010000001 to CBXY010037781), we found the sequences of nanos2 and gfra1a. The sequences were confirmed by PCR using testis cDNA and specific primers designed to the respective genomic sequences, showing open reading frames of 531 bp and 1419 bp coding for 177 and 473 amino acid residues, respectively. Although tissue specificity analysis by non-quantitative RT-PCR showed expression in all tissues studied, nanos2 expression was higher in testis, while gfra1a was highly expressed in gonads, brain and pituitary. Expression analysis by qrtPCR in testis during the reproductive cycle, showed the highest levels of nanos2 expression in testes containing only SpgA, decreasing as development was progressing. However, levels of gfra1a expression were high during early stages of development until spermatocytes were observed, decreasing afterwards. These results suggest that both genes can have key roles during the early stages of testicular development.
Spermatogenesis starts when undifferentiated type A spermatogonia (und-SpgA), also known as spermatogonial stem cells (SSC), become differentiated SpgA (diff-SpgA). Expression of nanos2 in und-SpgA has been observed in several species and it is recognized as a spermatogonial stem cell marker. In the mammalian testis, it is well established that Sertoli cells produce GDNF (glial cell line-derived neurotrophic factor) that binds the GFRA1 (GDNF family receptor alpha-1), which is expressed by und-SpgA promoting its self-renewal and maintenance. A few recent studies have addressed the potential involvement of the Gdnf-Gfra1 signaling pathway in the regulation of gametogenesis in cartilaginous and bony fishes. As a first step to determine nanos2 and gfra1a involvement in the self-renewal and maintenance of the und-SpgA preventing spermatogonial differentiation in the European seabass, the aim of this work was to characterize and study their expression at different stages of testicular development. By blast analysis against the NCBI wholegenome shotgun contigs (wgs) database for D. labrax (sequences CBXY010000001 to CBXY010037781), we found the sequences of nanos2 and gfra1a. The sequences were confirmed by PCR using testis cDNA and specific primers designed to the respective genomic sequences, showing open reading frames of 531 bp and 1419 bp coding for 177 and 473 amino acid residues, respectively. Although tissue specificity analysis by non-quantitative RT-PCR showed expression in all tissues studied, nanos2 expression was higher in testis, while gfra1a was highly expressed in gonads, brain and pituitary. Expression analysis by qrtPCR in testis during the reproductive cycle, showed the highest levels of nanos2 expression in testes containing only SpgA, decreasing as development was progressing. However, levels of gfra1a expression were high during early stages of development until spermatocytes were observed, decreasing afterwards. These results suggest that both genes can have key roles during the early stages of testicular development.
DescriptionTrabajo presentado en el XII Conference of the Iberian Association for Comparative Endocrinology (AIEC), celebrado en Faro (Portugal), del 26 al 28 de septiembre de 2019
URIhttp://hdl.handle.net/10261/209404
Appears in Collections:(ICMAN) Comunicaciones congresos
(IATS) Comunicaciones congresos
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