Project description:Regulation of presenilin genes. Presenilins are intramembrane aspartic proteases. These proteases are critical proteins in pathogenesis of Alzheimer's disease. The function of recently identified presenilin-homologous proteases (IMPAS or SPP)s is unknown. Our preliminary data in C.elegans model suggested the role of these proteins in early-development and , perhaps, in pathway related- to cholesterol -regulated signalling (Grigorenko et al, 2004, PNAS). The overall goal is to determine pattern of gene expression alterations in cells with knock-out or knock-down presenilin related proteins (IMPAS). The cultured cells, C.elegans and mouse models are planned to be used in this study. Here , we will determine gene expression patterns in C.elegans strain NL2099, fed with bacteria HT115, producing dsRNA for Ce-imp-2 gene (knock-down of imp-2 gene by RNAi )or GFP gene as a control. We hypothesize that novel family of presenilin-related proteases is important for cholesterol-regulated intracellular signalling and early development (including CNS/neuronal development and function). C.elegans (strain NL2099) on L1 stage were placed on agarose plates with bacteria HT115, producing dsRNA for: 1) Ce-imp-2 gene; 2) GFP as a control gene. After 48 hours at 20C , worms were washed in M9 buffer and total RNA was isolated by TRIzol method.
