TY - JOUR
T1 - The use of real-time PCR analysis in a gene expression study of Alzheimer's disease post-mortem brains
AU - Gutala, Ramana V.
AU - Reddy, P. Hemachandra
N1 - Funding Information:
The authors thank the Harvard Tissue Resource Center, Belmont, MA (which is supported in part by PHS Grant MH/NS 31862), for providing the brain specimens and the necessary pathological information. The authors also thank Geoffrey Murdoch, MD, Department of Pathology, OHSU for pathological interpretation of the brain specimens. This research was supported in part by the Alzheimer’s Association of Oregon, the Medical Research Foundation of Oregon, the American Federation for Aging Research, and the Alzheimer’s Disease Center of Oregon, NIA Grant P30 AG08017.
PY - 2004/1/15
Y1 - 2004/1/15
N2 - The measurement of gene expressions in brains with neurodegenerative diseases is a major area of brain research. The objective of our research was to determine whether quantitative real-time PCR could measure messenger RNA (mRNA) expression in brains with post-mortem intervals beyond 12h. In the present paper, we examined the quality of RNA from brain specimens of both Alzheimer's disease (AD) patients (n=13) and non-demented normal control subjects (n=6). To determine a unregulated endogenous reference gene in AD, we measured mRNA expressions of the commonly used reference genes β-actin, 18S rRNA, and GAPDH. In addition, we determined whether post-mortem interval, brain weight, or age at death influences mRNA expression. Our real-time PCR analysis results indicate that mRNA expression can be detected in all brain specimens for β-actin, 18S rRNA, GAPDH, and also synaptophysin, a known marker for AD. Further, using real-time PCR analysis, we found that β-actin and 18S rRNA are differentially expressed in the brain specimens of both AD and control subjects, while GAPDH is similarly expressed in AD and control brain specimens. These findings suggest that GAPDH can be used as a endogenous reference gene in the study of AD brains. A comparative gene expression analysis also suggests that synaptophysin is down-regulated in AD brain specimens compared to control brain specimens.
AB - The measurement of gene expressions in brains with neurodegenerative diseases is a major area of brain research. The objective of our research was to determine whether quantitative real-time PCR could measure messenger RNA (mRNA) expression in brains with post-mortem intervals beyond 12h. In the present paper, we examined the quality of RNA from brain specimens of both Alzheimer's disease (AD) patients (n=13) and non-demented normal control subjects (n=6). To determine a unregulated endogenous reference gene in AD, we measured mRNA expressions of the commonly used reference genes β-actin, 18S rRNA, and GAPDH. In addition, we determined whether post-mortem interval, brain weight, or age at death influences mRNA expression. Our real-time PCR analysis results indicate that mRNA expression can be detected in all brain specimens for β-actin, 18S rRNA, GAPDH, and also synaptophysin, a known marker for AD. Further, using real-time PCR analysis, we found that β-actin and 18S rRNA are differentially expressed in the brain specimens of both AD and control subjects, while GAPDH is similarly expressed in AD and control brain specimens. These findings suggest that GAPDH can be used as a endogenous reference gene in the study of AD brains. A comparative gene expression analysis also suggests that synaptophysin is down-regulated in AD brain specimens compared to control brain specimens.
KW - Alzheimer's disease
KW - GAPDH
KW - Gene expression
KW - Post-mortem brains
KW - Real-time PCR
KW - β-Actin
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U2 - 10.1016/j.jneumeth.2003.09.005
DO - 10.1016/j.jneumeth.2003.09.005
M3 - Article
C2 - 14687679
AN - SCOPUS:0346995416
VL - 132
SP - 101
EP - 107
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
SN - 0165-0270
IS - 1
ER -