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Comparative analysis of testis transcriptome between a genetic male sterile line (GMS) and its wild-type 898WB in silkworm, Bombyx mori

ABSTRACT: Purpose:The purpose of study these processes, is helpful to understand the mechanisms of genetic sterility of lepidopteran insects and achieve sterile insect technique (SIT) for pest control. Methods:Intact testes were dissected on day 5 of fifth-instar larvae and immediately frozen in liquid nitrogen for RNA-Seq. The testes from three silkworms were taken as one biological sample, and three samples per strain were prepared as biological replicates. Results: A total of 1,872 up-regulated genes and 1,823 down-regulated genes were detected in the testis of the GMS mutant. Several genes play important roles in spermatogenesis and testicular development, such as “serine/threonine protein kinase”, “organic cation transporter protein”, “tyrosine protein kinase”,“lncRNAs” and “immune-associated genes”, and so on. GO (Gene ontology) annotation and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis was performed for functional annotation of the differentially expressed genes (DEGs). KEGG pathway analysis showed that the DEGs annotated to 123 pathways, and 10 pathways were enriched significantly, such as “metabolic pathway”, “biosynthesis of amino acids”, and “phagosome–lysosome pathway” associated with the testis development and spermatogenesis. The qPCR expression results were similar to the results obtained from the RNA-seq data, which showed the sequencing results were reliable. Conclusions:Differentially expressed genes of the testes between GMS mutant and 898WB were screened by RNA-Seq technology, which provide reliable reference for understanding the mechanism of male sterility in GMS mutant.

ORGANISM(S): Bombyx mori

PROVIDER: GSE191033 | GEO | 2021/12/17

REPOSITORIES: GEO

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Project description:The Drosophila spermatogenesis cell differentiation pathway involves the activation of a large set of genes in primary spermatocytes. Most of these genes are activated by testis-specific TATA-binding protein associated factors (tTAFs). In the current model for the activation mechanism, Polycomb plays a key role silencing these genes in the germline precursors, and tTAF-dependent activation in primary spermatocytes involves the displacement of Polycomb from gene promoters. We investigated the genome-wide binding of Polycomb in wild type and tTAF mutant testes. According to the model we expected to see a clear enhancement in Polycomb binding at tTAF-dependent spermatogenesis genes in tTAF mutant testes. However, we find little evidence for such an enhancement in tTAF mutant testes compared to wild type. To avoid problems arising from cellular heterogeneity in whole testis analysis, we further tested the model by analysing Polycomb binding in purified germline precursors, representing cells before tTAF-dependent gene activation. Although we find Polycomb associated with its canonical targets, we find little or no evidence of Polycomb at spermatogenesis genes. The lack of Polycomb at tTAF-dependent spermatogenesis genes in precursor cells argues against a model where Polycomb displacement is the mechanism of spermatogenesis gene activation. This genome-wide ChIP-array study investigates the binding of Polycomb in three biological samples: wild type (WT) whole testes, tTAF (can) mutant whole testes, and FACS-sorted germline precursor cells. We performed two biological replicates for each sample, except wild type whole testes where we performed three. For all ChIP-array experiments, input chromatin was used as the reference control to assay ChIP enrichment. We used Cy3/Cy5-labelled ChIP and input DNA for hybridisation onto Nimblegen arrays, and we performed a Cy3/Cy5 dye swap for one biological replicate of each sample (see supplementary file: GSE39935_README.txt).

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Comparative analysis of testis transcriptome between a genetic male sterile line (GMS) and its wild-type 898WB in silkworm, Bombyx mori

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