The Role of TRPC1 on Early-Onset Alzheimer’s Disease
Project no. 21058 · Status: Kapanmış (Yayın bekleniyor) · 2020
Summary
Aim of the project was to investigate the store-operated calcium entry (SOCE), expression levels of SOCE-associated genes, Aß protein levels and proliferation rate on cells that have Early-Onset Alzheimer’s Disease (EOAD)-associated presenilin-1 (PSEN1) mutation and overexpress TRPC1. Mutant PSEN1 SH-SY5Y cells were transfected with TRPC1 cDNA containing overexpression plasmids to investigate the effects of TRPC1. Aß levels were determined by ELISA and western blot. SOCE was monitoring on cells that are loaded with Ca2+-indicator Fura-2 by sprectrofluorometry real-timely. Expression levels of SOCE-associated genes were determined with RT-qPCR. Proliferation rates of cells were monitoring with real-time cell analysis system. While Aß levels increased at mtPSEN1 cells compared to control cells statistically significant, SOCE decreased. The mutation did not cause any effect on the expression levels of STIM1 and TRPC1 whereas TRPC6 and Orai1 decreased. A significant increase was observed with TRPC1 overexpression. Expression levels of TRPC6, STIM1 and Orai1 genes at TRPC1 overexpress mutant cells were similar to mutant cells. Aß levels, which increased with mutation, decreased statistically significant in mtPSEN1+TRPC1 cells compared to mutant cells. Both of mutation and TRPC1 overexpression caused a decrease in proliferation rate. Alterations of SOCE and SOCE-associated genes expression levels demonstrate that these proteins and their mediated SOCE may have a role on AD pathology. At the same time, reduction of Aß levels with TRPC1 overexpression estimated that TRPC1 may be a new therapeutical target and contribute to the development of new drugs for Alzheimer disease. But reduction of Aß levels could not treat the AD’s complex pathology. As a consequence, further in vitro and in vivo experiments should be achieved with activatory/inhibitory agents of SOCE for illuminating of mechanism and therapeutic effects.