Polyunsaturated fatty acids as possible modulators of vanilloid signalling in human neuroblastoma

• Grant number: 12/18522-0
• Support Opportunities: Regular Research Grants
• Duration: January 01, 2013 - December 31, 2014
• Field of knowledge: Biological Sciences - Physiology - Physiology of Organs and Systems
• Principal Investigator: Luiz Roberto Giorgetti de Britto
• Grantee: Luiz Roberto Giorgetti de Britto
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Recent data of our laboratory indicate that the activation of TRPV1 channels induces apoptotic cell death during the development of the retina, which is partially dependent on nitrosative stress. Oxidative and endoplasmic reticulum stress may be also involved in these processes. Our preliminary data indicate that human retinoblastoma Y79 cells express TRPV1 channels, and that their activation cause apoptosis. However, although it is known that Y79 cells are extremely susceptible to polyunsaturated fatty acids-induced cell death, the function of TRPV1 channels in these processes remains to be elucidated. In this project, we intend to expose Y79 cells to increasing concentrations of polyunsaturated fatty acids and TRPV1 agonists and antagonists. Cell viability, DNA fragmentation, cell proliferation, and superoxide and nitric oxide formation will be assessed by flow cytometry. We are also going to evaluate the nitrosilation of some proteins that are involved in the generation of reactive oxygen species. Gene and protein expression of several molecules that are involved in endoplasmic reticulum stress and redox balance will be assessed by real time rt-PCR and immunoblotting. Confocal microscopy will be used to evaluate membrane migration of several NADPH oxidase complex proteins. We anticipate that the results could contribute data to the knowledge of the mechanisms involved in retinal development and to retinoblastoma prevention and treatment. (AU)