Advanced search
Start date
Betweenand

Next-generation mRNA sequencing for high-throughput analysis of the metamorphosis in Apis mellifera castes

Grant number: 12/12941-1
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): October 01, 2012
Effective date (End): January 31, 2016
Field of knowledge:Biological Sciences - Genetics
Principal Investigator:Marcia Maria Gentile Bitondi
Grantee:Michelle Prioli Miranda Soares
Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:11/03171-5 - Causal analysis of Apis mellifera development - regulatory genes and hierachical networks of gene expression in the specification of tissue and organs, AP.TEM

Abstract

In the broadest sense, metamorphosis is a developmental process by which an immature organism is transformed into a sexually mature adult. In insects, this process is characterized by the degeneration, or remodeling, of larval tissues and formation of the adult tissues from nests of precursor cells localized to specific regions of the larval body. To our knowledge, large-scale analysis of gene expression during metamorphosis was performed only in a model insect, Drosophila, by using microarrays composed of cDNA sequences representing only 30% to 40% of the estimated number of genes in the respective genome. Here, we propose a more comprehensive analysis of gene expression variation during metamorphosis. To this end, our purpose is to use the RNA seq (high-throughput RNA sequencing) methodology for sequencing the transcriptome of the honey bee (Apis mellifera) in the course of metamorphosis. With deep coverage and base-level resolution, the next-generation sequencing allows screening the differentially expressed genes during precise stages of metamorphosis, besides providing accurate measurements of transcript levels. Following sequencing, the resulting reads will be mapped to the genome of the honey bee. Besides capturing transcriptome dynamics across metamorphosis, a series of bioinformatics tools will enable: (1) to group these genes into classes according to their potential functions, (2) the searching for common sites in the gene promoter regions that could indicate regulation by transcription factors or hormones, thus allowing the construction of co-regulatory networks, (3) to establish the hierarchical gene expression networks acting during metamorphosis. Furthermore, we are proposing the use of workers and queens in this project in order to investigate caste-specific gene activity during metamorphosis. It is hoped that this approach reveals the wide panorama of the molecular genetics of metamorphosis.