Proliferation and differentiation of cerebellar neural stem cells (CNSC) in response to SHH and CXCL12 treatment

• Grant number: 16/25400-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2017
• End date: December 31, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Marimélia Aparecida Porcionatto
• Grantee: Giovana Longatti Baptista
• Host Institution: Instituto Nacional de Farmacologia (INFAR). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

The cerebellum is responsible for controlling different motor and cognitive functions. It is also an important model for studying neurogenesis because it has a simplified cortical architecture and a considerable part of its development unravels after the embryonic stage. Cerebellar neural stem cells (CNSC) that can differentiate into the main cell types of the cerebellum have been isolated from postnatal and adult mice. The present study aims to characterize CNSC extracted from new-born and adult mice according to their proliferation, self-renewal and differentiation responses to treatment with Sonic hedgehog (Shh), one of the most important mitogens during the development of the nervous system, and with CXCL12, a cytokine which interacts with the Shh signalling pathway and has a role in neural precursor migration. We will extract CNSC from P6 and P45 C57/black mice and quantify Shh and CXCL12 receptors (Smoothened, Patched, CXCR4 and CXCR7) using quantitative PCR (qPCR). After combined or isolated treatment in vitro with Shh and CXCL12, we will evaluate CNSC rates of proliferation and self-renewal by counting and measuring the generated neurospheres and its differentiation profile using immunocytochemistry. Proliferation, self-renewal and differentiation will also be evaluated after pre-treatment assays, in which the CNSC will receive CXCL12 before the treatment with Shh. The described experiments will show the effects of Shh on in vitro CNSC proliferation and differentiation, and if CXCL12 is a necessary and/or conditioning factor for enabling Shh actions in these cells. (AU)