Identification of membrane proteins in Apis mellifera brain submitted to the proboscis extension reflex assay

• Grant number: 15/26025-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: March 01, 2016
• End date: February 28, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Mario Sergio Palma
• Grantee: Anally Ribeiro da Silva Menegasso
• Supervisor: Gert Lubec
• Host Institution: Instituto de Biociências (IB). Universidade Estadual Paulista (UNESP). Campus de Rio Claro. Rio Claro , SP, Brazil
• Institution abroad: University of Vienna, Austria
• Associated to the scholarship: 13/13542-6 - A neuroproteomic approach of the brain from Africanized Apis mellifera submitted to the assay proboscis extension reflex, BP.DR

Abstract

We are studying the bees brain's response to a stimuli that lead to execution of unconditional behaviors. The response of neurons to this type of stimuli is of biochemical/neurochemical nature, in which many of the physiological responses are carried by nervous receptors and/or ion channel (and/or its analogues), which mostly are located in the level of neuron's membrane. The classical proteomics identifies well soluble proteins, which are usually located in the cytoplasm, and sometimes proteins non-covalently linked to nerve receptors (but this situation is a minority of the proteins). This means that under the biological point of view, the results we have obtained so far explain only part of the adaptive bee brain responses to unconditioned stimuli. In the present study the behavior of "Proboscis Extension Reflex" (PER) will be used as input reference for membrane proteome studies of bee brain neurons. For this approach to following experimental strategies will be used: i) proteome analysis of membrane proteins complexes (nerve receptors) by native electrophoresis (BN-PAGE) followed by mass spectrometry analysis using a LC-MS/IT-ToF system of honeybees brain subject to PER test; ii) proteome analysis of membrane proteins complexes by gel-free followed by shotgun proteomics using a LC-MS/IT-ToF system of honeybees brain subject to PER test and iii) analysis of PTMs assignment using a LC-IT-ETD system. To develop this type of proteomic analysis is required mastery of delicate cell membranes manipulation protocols, and especially training of highly qualified human resources, which we do not have in Brazil. The identification of nerve receptors and ion channels in the neuronal membrane level, as well the PTMs analysis may contribute to better understand the cognitive bases in honeybee.