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Study of meiotic X chromosome inactivation in Drosophila spp. spermatogenesis

Grant number: 19/15212-0
Support type:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): October 01, 2019
Effective date (End): January 31, 2022
Field of knowledge:Biological Sciences - Genetics
Principal Investigator:Maria Dulcetti Vibranovski
Grantee:Camila Correia Avelino
Home Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Associated research grant:15/20844-4 - The role of gametogenesis on the origin and evolution of new genes, AP.JP

Abstract

Meiotic Sex Chromosome Inactivation (MSCI) is a process of transcriptional silencing of the sex chromosomes in the early stages of meiosis in spermatogenesis. Scientists have investigated the existence and impact of MSCI on D. melanogaster and some aspects related to spermatogenesis raised a great debate about the existence of this mechanism. During the period of the master's degree, the candidate participated in a collaboration with the laboratory of Professor Brian Oliver (NIDDK - NIH), and they found by Single-Cell RNA-seq a reduction of the X and IV expression, the IV chromosome was previously described in the literature as an ancestral X chromosome. In addition, the candidate performed cytogenetic experiments (Double Immuno-FISH) that found evidence of a depletion of RNA polymerase II on X and IV chromosomes in the late stages of meiotic prophase I, compared to the autosomes. As the double Immuno-FISH is a qualitative result, it is not possible to verify if the same genes that suffered a reduction of expression are those that did not show binding with RNA polymerase II in the late phases of prophase I. In order to investigate how MSCI mechanism occurs in its ancestral aspect and analyze gene regulation in Drosophila, this project intends to use three approaches: Double Immuno-FISH, for qualitative verification of MSCI in D. melanogaster; ChIP-seq, as a quantitative verification of the phenomenon observed by the Immuno-FISH and finally the differential expression analyses of RNA-seq data of the three main phases of spermatogenesis (mitosis, meiosis and post meiosis) from three species of the genus Drosophila, in order to observe the MSCI panorama in different evolutionary stages. Therefore, it is expected that these results will contribute to the comprehension of the evolution and regulation of MSCI in Drosophila male germline. (AU)