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Transcriptome of tambaqui Colossoma macropomum during gonad differentiation: different molecular signals leading to sex identity

ABSTRACT: Purpose: The gene molecular network involved in teleost fish sex determination and differentiation is highly variable among species and even in some cases among populations of the same species. The objectives of the present study were to identify the period of gonadal sex differentiation in tambaqui juveniles, as well as the genes and pathways potentially involved in this process. Methods: Histological analysis of juveniles was carried out to establish a timeline of the gonadal differentiation in tambaqui. Based on that knowledge, ten juveniles were selected before the first evidence of histological sex differentiation and total RNA was extracted from their trunks and used for RNA-Sequencing and a subsequent de novo transcriptome assembly. Principal Component Analysis (PCA) of the whole transcriptome data was used to cluster samples into two distinct groups: putative males and putative females. Differential gene expression, functional annotation and gene enrichment were used to identify genes and pathways related to sex differentiation in tambaqui to which was applied the Mann-Whitney non-parametric t test (p <0.05) confirming the statistical significance of the expression dimorphism between the groups. Results: The first sign of histological sex differentiation in tambaqui was the formation of the ovarian cavity detected in individuals measuring about 40 mm in total length. Before the differentiation period, components of the Wnt / β-catenin pathway, fox and fst genes (p <0.05) suggest female sex development in the putative females, whereas antagonistic pathways (gsk3b, wt1 and fgfr2), sox9 and genes for androgen synthesis (p <0.05) are indicative of a male-like differentiation. Conclusions: Tambaqui juveniles prior to the morphological ovarian differentiation present the Wnt / β-catenin pathway exerting putative role on the sex differentiation target, either upregulated in female-like individuals, or antagonized in male-like individuals. Thus, the present work provides a molecular basis for future studies on the application of tambaqui monosex cultivation.

ORGANISM(S): Colossoma macropomum

PROVIDER: GSE130895 | GEO | 2020/08/05

REPOSITORIES: GEO

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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.

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Transcriptome of tambaqui Colossoma macropomum during gonad differentiation: different molecular signals leading to sex identity

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