Correlation of the expression level of C1q mRNA and the number of C1q-positive plaques in the Alzheimer Disease temporal cortex. analysis of C1q mrna and its protein using adjacent or nearby sections.
We compared the expression level of C1q mRNA and the number of C1q-positive plaques in adjacent or nearby brain sections from Alzheimer disease (AD) and control cases. Small blocks of temporal cortex were fixed with 4% paraformaldehyde for 2 days at 4 degrees C. After cryoprotection with solutions containing 10-20% glycerol and 2% dimethylsulfoxide, 40-microm sections were cut from the tissue blocks. A section from each case was stained by immunohistochemistry using a C1q antibody, while RNA was purified from adjacent or nearby sections using a combination of proteinase K pretreatment followed by extraction using Trizol reagent. The expression of C1q B chain mRNA was analyzed in these samples by the reverse-transcription polymerase chain reaction (RT-PCR). The intensities of the PCR products were measured by an image analyzer. The expression of C1q B chain mRNA was significantly more abundant in AD than in control cases (p < 0.05). Immunohistochemical analysis showed that C1q protein was localized in senile plaques in the AD brain. The number of C1q-positive plaques correlated with the expression level of C1q gene (p < 0.05). The present results suggest that C1q protein in senile plaques originates is endogenously produced in the AD brain.
Tooyama
I
I
Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu, Japan. kinchan@sums.shiga-med.ac.jp
Sato
H
H
Yasuhara
O
O
Kimura
H
H
Konishi
Y
Y
Shen
Y
Y
Walker
D G
DG
Beach
T G
TG
Sue
L I
LI
Rogers
J
J
D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D001923:Brain Chemistry; D016022:Case-Control Studies; D015922:Complement C1q / Q000096:biosynthesis* / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D058225:Plaque, Amyloid / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013702:Temporal Lobe / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*
D012333:RNA, Messenger; D015922:Complement C1q