Identification and characterization of miRNA-target interactions in the metamorphosis of Apis mellifera

• Grant number: 14/18091-5
• Support Opportunities: Scholarships in Brazil - Master
• Start date: December 01, 2014
• End date: January 31, 2016
• Field of knowledge: Biological Sciences - Genetics - Animal Genetics
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Zilá Luz Paulino Simões
• Grantee: Natália Helena Hernandes
• Host 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

Molting and metamorphosis of insects are mainly driven by a balance of two hormones: Ecdysteroids (20E) and Juvenile Hormone (JH). Such hormones coordinate the succession of developmental stages through the regulation of specific genes. Ecdysteroids stimulate biochemical and morphological events during molting cycle and JH directs the action of ecdysteroids in the development. During the larval stage, when the animal reaches the characteristic species size, the JH levels decrease; as a consequence, the larvae go through transformation processes culminating in metamorphosis. JH titles are determined by the balance of the synthesis in the corpora allata and the degradation by the Juvenile Hormone Esterase (JHE). Several genes are involved in the downstream pathway of both hormones, like Methoprene-tolerant (MET), Ecdysone Receptor (ECR), Ultraspiracle (USP) and Calponin (CAL), although their specific functions remain unknown. MET mediates the action of JH by activating an early target, Krüppel homolog 1 (Kr-h1), whose function is to inhibit premature metamorphosis. These early genes of the endocrine network coordinate the activation of late genes, which regulate programmed cell death in larval tissues, central nervous system remodeling, imaginal discs proliferation and morphogenesis. In addition to the neuroendocrine system, the process of metamorphosis is controlled by nutritional pathways (insulin) and by regulators such as microRNAs. Apis mellifera is an excellent model for developmental biology studies as a diploid egg can give rise to both workers and queens, which makes possible to address questions related to genetic metamorphosis, a field that remains to be exploited. Thus, using the information available and our databases, our goal is to identify and characterize the targets of miR-34, miR-252a, miR-252b and miR-281, known as important regulators of insect metamorphosis. For this purpose, we propose 1) the use of predictors to identify putative miRNA-target interactions and 2) validation of predicted miRNA-target interactions involving genes with role in metamorphosis: CAL, FTZ-f1, Kr-h1, JHE, INR, USP by luciferase assay in COS-7 cells. (AU)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

HERNANDES, Natália Helena. Identification and characterization of miRNA-target interactions in the metamorphosis of Apis mellifera. 2016. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC) Ribeirão Preto.