Project description:In Hymenoptera, sex is determined by a single-locus complementary sex determining system (sl-CSD). Males are normally haploid (hemizygous at the sl-CSD locus) but if the genotype of the sl-CSD locus is homozygous, they develop into diploid males. Here, we study the effects of ploidy and the sl-CSD-locus genotype by comparing gene expression differences between haploid males, diploid males and virgin queens at three developmental stages, pupae, 1 day and 11 days after eclosion. Keywords: diploid males, sl-CSD, ploidy-spcific gene, sex-specific gene, doublesex, sex-biased gene
Project description:In Hymenoptera, sex is determined by a single-locus complementary sex determining system (sl-CSD). Males are normally haploid (hemizygous at the sl-CSD locus) but if the genotype of the sl-CSD locus is homozygous, they develop into diploid males. Here, we study the effects of ploidy and the sl-CSD-locus genotype by comparing gene expression differences between haploid males, diploid males and virgin queens at three developmental stages, pupae, 1 day and 11 days after eclosion. Keywords: diploid males, sl-CSD, ploidy-spcific gene, sex-specific gene, doublesex, sex-biased gene
Project description:In Hymenoptera, sex is determined by a single-locus complementary sex determining system (sl-CSD). Males are normally haploid (hemizygous at the sl-CSD locus) but if the genotype of the sl-CSD locus is homozygous, they develop into diploid males. Here, we study the effects of ploidy and the sl-CSD-locus genotype by comparing gene expression differences between haploid males, diploid males and virgin queens at three developmental stages, pupae, 1 day and 11 days after eclosion. Keywords: diploid males, sl-CSD, ploidy-spcific gene, sex-specific gene, doublesex, sex-biased gene
Project description:PBANKA_0828000 was identified in a screen for sex determination genes in Plasmodium berghei. PBANKA_0828000 encodes a zinc finger protein of unknown function. To address its role, PBANKA_0828000 was tagged with an HA tag at the endogenous locus, and protein interactions were characterised by affinity purification followed by mass spectrometry.
Project description:Purpose: Determine whether sex-determining genes are bivalent at the bipotential stage, poised between the testis and ovary fate, and whether H3K4me3 and H3K27me3 resolve into sex-specific patterns after sex determination, contributing to the canalization and stabilization of either the testis or ovary fate. Methods: XX and XY supporting cells of the gonad were FACS-purified before sex determination (at E10.5) and after sex determination (at E13.5), and submitted to ChIP-seq for H3K4me3, H3K27me3 and H3 as a means to normalize across cell populations. Results: We found that key sex-determining genes are bivalent at the bipotential stage. Genes that are upregulated affter sex determination are stripped of their repressive H3K27me3 mark, whereas repressed genes that promote the alternate pathway remain bivalent even after sex determination.
2019-05-07 | GSE130749 | GEO
Project description:Cryptobranchid sex determination
Project description:Sex determination mechanisms are bewilderingly diverse. A cascade of genes with hierarchical regulation characterizes sex determination pathways in insects. How such pathways evolve is poorly understood, partly due to a lack of comparative data. Houseflies are well known for their polymorphic sex determination with populations carrying a dominant male determiner M on the Y chromosome or any of the five autosomes. We identified the male determining gene Mdm responsible for splicing regulation of the key switch transformer by exploiting the existence of housefly populations with different sex determination mechanisms. We demonstrate that Mdm originated from duplication of CWC22/nucampholin, a generic and essential splicing regulator across Metazoans with a crucial role in exon-junction complex assembly and non-sense mediated decay. We show that strains with the M-locus on the Y and on different autosomes carry multiple copies of Mdm indicating that the same male determiner translocated to different genomic sites in the genome. We found that embryonic RNAi-based silencing of Mdm leads to differentiation of ovaries in males, while targeted Mdm disruption with CRISPR/CAS-9 resulted in complete sex reversal to fertile females. Our study reveals how a duplicate of a gene with a general splice-regulation role during development can be recruited to serve a specific function in the determination of male sex. Mdm appears to be unique to the housefly representing a compelling example for the plasticity at the instructive level of sex determination hierarchies.
Project description:Purpose: In this study we employed unbiased, genome-wide techniques to identify regulatory elements during murine sex determination. Methods: We performed ATAC-seq on 60K FACS-purified XX and XY gonadal cells before and after sex determination to map nucleosome depleted regions (NDRs) indicative of regulatory elements. To determine whether these are active enhancers, we performed ChIP-seq for H3K27ac, a histone modification that marks active enhancers in both sexes and time points. Transient transgenics was performed on select enhancers to determine whether they are functional in gonads during the sex determination stage. Results: We have produced a genome wide map of potential regulatory elements and active enhancers during the process of murine sex determination. Furthermore, we validated the power of our dataset by identifying a novel enhancer downstream of Bmp2, a female-specific gene. Conclusions: This work supplies a powerful resource for identifying chromatin regulatory elements active during mammalian sex determination.