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Identification and validation of miRNA-mRNA interactions during preimaginal phases of Apis mellifera

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Author(s):
Thiago da Silva Depintor
Total Authors: 1
Document type: Master's Dissertation
Press: Ribeirão Preto.
Institution: Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC)
Defense date:
Examining board members:
Zila Luz Paulino Simoes; David de Jong
Advisor: Zila Luz Paulino Simoes
Abstract

The development in honeybees is mainly controlled by the action of two major hormones, juvenile hormone (JH) and 20-hidroxyecdysone (20E). These hormones trigger gene cascades, which results in phenotypic, physiological and behavioral changes. Besides hormones, a class of non-coding RNAs, the microRNAs, regulates gene expression at a post-transcriptional level during insect development. In this study we aimed to analyze the relationship between developmental genes and morphogenetic hormones, in final stages of the development of Apis mellifera. The expression profile of the orphan nuclear receptor Usp, ftz-f1, EcR, chd64, inr2, Kr-h1), gce, early-trypsin, and their putative regulators miRNA-34, miRNA-281, miRNA-252a and miRNA-252b were assessed from 5? instar larvae to newly emerged adults by qPCR. The effect of exogenous doses of both hormones applied on white eyed pupae (Pw) and brown eyed pupae (Pb) was also tested. Most of the genes seem to respond to hormonal variation in pupal stages as they do in larval stages. However, gce and chd64 showed a different response to hormonal treatment in pupal states, thus suggesting they play different roles in final stages of development. Unexpectedly gce, which is a nuclear receptor of JH in insects, showed a quick response to 20E treatment and no response to JH in pupal stages of honeybees, as well as chd64 which also responded only to the 20E treatment. In addition, we recognized Usp as an Immediate Early Gene, for it responded rapidly to hormonal treatments and quickly restored its level. In addition, we find the miR-34 and miR-281 as strong candidates of regulators since they presented many putative interactions in the 3\'UTR of the candidate genes and showed to be affected by the hormonal treatment. This study describes new components to the regulatory network that regulates bee development. (AU)