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Molecular Analysis of Intracellular Trafficking of the Voltage Gated Proton Channel by Fluorescence Microscopy

Grant number: 16/16346-1
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): March 01, 2017
Effective date (End): July 31, 2020
Field of knowledge:Biological Sciences - Physiology - Physiology of Organs and Systems
Principal Investigator:Manoel de Arcisio Miranda Filho
Grantee:Luísa Ribeiro Silva
Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

The voltage gated proton channel (HV1) plays an important role during microbial killing by phagocytes, such as neutrophils and microglia. Besides its physiological functions, recent studies have shown that HV1 channel participates in pathological states, not only increasing tissue damage in ischemic stroke, but also promoting the proliferation and survival of tumoral cells. Even though these pathological processes are associated with an augmented surface expression of the HV1 channel, little is known about the cellular and molecular mechanisms that determine its abundance at the plasmatic membrane.The superficial expression of ion channels is defined, principally, by three processes; protein synthesis, exocytosis and endocytosis of the channel at the plasmatic membrane, and protein degradation. These processes are finally regulated and, generally, depend upon signals contained in the intracellular domain of the ion channels.The present research project proposes to investigate the molecular mechanisms of the superficial expression of the voltage gated proton channel, associating modifications in its C-terminal domain with its intracellular localization. Through fluorescence microscopy and molecular biology techniques, we pretend to identify the secretory and endocytic pathway used by the Hv1 channel, and the relevance of its C-terminal domain to the channels trafficking.

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