Amyloid precursor protein heat shock response in lymphoblastoid cell lines bearing presenilin-1 mutations.
The amyloid precursor protein (APP) gene promoter contains a heat shock element. An abnormal APP heat shock response could increase accumulation of A beta, the APP metabolite found in Alzheimer's disease amyloid plaques. Since A beta production is affected by presenilin-1 (PS-1) mutations, we investigated whether basal APP levels or response to heat shock were altered in lymphoblastoid cell lines from 8 PS-1 mutation-bearers and 9 control members of Alzheimer's disease families. Lymphoblastoid cell lines were incubated at 42 degrees C for 35 min and allowed to recover at 37 degrees C for 1, 3, 8, 24 and 48 h. APP mRNA levels, quantified using RNA-RNA solution hybridisation, increased significantly at 1 and 3 h post-heat shock to between 123% and 163% of pre-heat shock (0 h) levels and returned to normal by 8 h. Semi-quantitative Western immunoblotting of cell lysates using the 22C11 antibody detected two major bands, migrating at approximately 145 and approximately 120 kDa. Band optical densities increased significantly at 3 h to approximately 155% of 0 h levels, following the increase in APP mRNA levels and showing a similar reversibility. APP mRNA and protein responses were comparable in the PS-1 mutation-bearing and control cell lines. This study shows that both APP mRNA and protein are induced in lymphoblastoid cell lines following heat shock and that this response is not affected by PS-1 mutations which are pathogenic for Alzheimer's disease.
Johnston
J A
JA
Karolinska Institute, Department of Clinical Neuroscience and Family Medicine, Novum KFC, Huddinge, Sweden. j.a.johnston@qub.ac.uk
Lannfelt
L
L
Wiehager
B
B
O'Neill
C
C
Cowburn
R F
RF
D000544:Alzheimer Disease / Q000235:genetics; D016564:Amyloid beta-Protein Precursor / Q000235:genetics*; D002460:Cell Line; D005786:Gene Expression Regulation; D018869:Heat-Shock Response; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D053764:Presenilin-1; D012333:RNA, Messenger / Q000235:genetics
D016564:Amyloid beta-Protein Precursor; D008565:Membrane Proteins; C508853:PSEN1 protein, human; D053764:Presenilin-1; D012333:RNA, Messenger