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Calcium binding protein regulation and retinal degeneration by mechanical trauma

Grant number: 12/03214-9
Support type:Scholarships in Brazil - Master
Effective date (Start): July 01, 2012
Effective date (End): May 31, 2013
Field of knowledge:Interdisciplinary Subjects
Principal Investigator:Alexandre Hiroaki Kihara
Grantee:Fausto Colla Cortesao Zuzarte
Home Institution: Centro de Matemática, Computação e Cognição (CMCC). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil

Abstract

Calcium binding protein regulation and retinal degeneration by mechanical traumaAbstractCalcium is an ion with many roles within the cell. Its influx plays an important part in the secretion of neurotransmitters in neuronal chemical synapse transmission. Moreover, intracellular concentrations determine cell cycle progression and differentiation. There is a growing interest in the role of calcium in cell death and survival, implied as a signaling ion for trauma induced retinal degeneration (Knöferlea, J. & Kocha, J., 2010), besides acting on the regulation of nitric oxide, associated to diabetic retinal degeneration in mice (Kim, Y. H., & Kim, Y. S., 2011).This project aims to describe intracellular calcium related systems. In particular, EF hands calcium binding proteins calbindin, calretinin and parvalbumin seem to have a role in de ion's intracellular buffering, thus avoiding its free form's harmful effects. Genic expression modulation and intracellular levels of these proteins will be subject of biochemical and histological methods such as real-time PCR, western blot and immunofluorescence. Cellular death will be evaluated by different methodologies both morphological (Tunel, FluoroJade, nuclear pigments such as propidium iodide and DAPI) and biochemical (LDH assay, agarose gel DNA fragmentation, caspase activation). The assays will be applied to traumatized retinas, both in explants and in vivo. For a study in calcium dynamics, chelating agents such as EDTA, EGTA and BAPTA will be used. The equipment and supplies used in this research are financed by FAPESP (Project Junior Researcher "Cellular Docking in the Arch of Life: Development, Adaptation and Degeneration of the Central Nervous System", 2008/55219-1) and MCT/CNPq (Universal Edict, "A new look over cellular docking: development and degeneration of the visual system", 482859/2009-1), both conducted by Prof. Alexandre H. Kihara, PhD.