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RNA-sequencing and function analysis of adult gonad tissue identify potential key transcription factors in Macrobrachium resenbergii with sex determination

ABSTRACT: M. rosenbergii is a classical aquatic organism with sex-reversed phenomenon by gene-expressed regulation. Reversal of gonad development in M. rosenbergii received great attention but the role of transcription factors for gonadal maintenance and sex determination in adult shrimps remain unclearly. In this study, we sequencing the transcriptomes for the adult testis, ovary and androgenic gland by third-generation sequencing plus second-generation sequencing, and 21,345 were identified. In total, 9,199 differentially expressed genes (DEGs) were identified in pairwise comparison where 2,990 DEGs were upregulated expression in testis than other tissues, and 5,775 DEGs were upregulated expression in ovary, and 2,244 DEGs were upregulated expression in androgenic. There are 272 differentially expressed transcription factors (DETs) in total. The upregulated DETs in one of the testis, ovary and androgenic glands compared to the others were 107, 152 and 13 respectively. The GO and KEGG of DETs analysis suggested that MAPK signaling pathway and transcriptional misregulation pathway have key function in testis, the thyroid hormone signaling pathway and neuroactive ligand-receptor interaction play a important role in ovary. We determined the existence of networks containing significant DETs related to sex determination in the adult goands of M. rosenbergii. The key DETs are Piwi (only expression in testis and ovary) and Argonaute 2 (only expression in ovary) might be involved with the regulation of ovary development.

ORGANISM(S): Macrobrachium rosenbergii

PROVIDER: GSE171823 | GEO | 2021/04/11

REPOSITORIES: GEO

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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. Experiment Overall Design: RNA samples from two different groups of 20-40 pooled gonads for each sample. Embrionic testis and ovaries of age E13, E14, or E16, and E13 testis cultured for three days were compared to each other

Project description:The current study investigates the direct effects of in utero vinclozolin exposure on the developing rat testis transcriptome. Vinclozolin is a commonly used fungicide in agriculture and is an endocrine disruptor with anti-androgenic activity. Previous studies have demonstrated that exposure to vinclozolin during embryonic gonadal sex determination induces epigenetic modifications of the germ line and transgenerational adult onset disease states that include spermatogenic cell defects, prostate disease, kidney disease, and tumor development. An investigation of the molecular actions of vinclozolin was initiated through an analysis of direct actions on the F1 generation embryonic testis development. Microarray analyses were performed to compare control and vinclozolin treated testis transcriptomes at embryonic day 13, 14 and 16. A total of 576 differentially expressed genes were identified and the major cellular functions and pathways associated with these altered transcripts were examined. The sets of regulated genes at the different development periods were found to be transiently altered and distinct. Interestingly, genes previously shown to be regulated during normal male sex determination were not altered by vinclozolin treatment. Categorization by major known functions of all 576 genes altered by in utero vinclozolin exposure demonstrates transcription, signaling, cytoskeletal and extra cellular matrix associated transcripts are highly represented. Specific cellular process and pathway analyses suggest the involvement of Wnt and calcium signaling, vascular development and epigenetic mechanisms as potential mediators of the direct F1 generation actions of vinclozolin. For Samples 1-12: We used microarrays to determine genes expressed differentially between control and in utero Vinclozolin treated E13, E14, and E16 rat testis. For Samples 13-16: We used microarrays to determine genes expressed differentially between control and in vitro Vinclozolin treated E13 cultured rat testis. For Samples 17-20: We used microarrays to determine genes expressed differentially between control and in vitro Flutamide treated rat E13 cultured testis.

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RNA-sequencing and function analysis of adult gonad tissue identify potential key transcription factors in Macrobrachium resenbergii with sex determination

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