The use of real-time PCR analysis in a gene expression study of Alzheimer's disease post-mortem brains

Ramana V. Gutala, P (Hemachandra) Reddy

    Research output: Contribution to journalArticle

    88 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish (US)
    Pages (from-to)101-107
    Number of pages7
    JournalJournal of Neuroscience Methods
    Volume132
    Issue number1
    DOIs
    StatePublished - Jan 15 2004

    Fingerprint

    Real-Time Polymerase Chain Reaction
    Alzheimer Disease
    Gene Expression
    Brain
    Actins
    Messenger RNA
    Synaptophysin
    Genes
    Research
    Neurodegenerative Diseases
    RNA
    Weights and Measures

    Keywords

    • β-Actin
    • Alzheimer's disease
    • GAPDH
    • Gene expression
    • Post-mortem brains
    • Real-time PCR

    ASJC Scopus subject areas

    • Neuroscience(all)

    Cite this

    The use of real-time PCR analysis in a gene expression study of Alzheimer's disease post-mortem brains. / Gutala, Ramana V.; Reddy, P (Hemachandra).

    In: Journal of Neuroscience Methods, Vol. 132, No. 1, 15.01.2004, p. 101-107.

    Research output: Contribution to journalArticle

    Gutala, Ramana V. ; Reddy, P (Hemachandra). / The use of real-time PCR analysis in a gene expression study of Alzheimer's disease post-mortem brains. In: Journal of Neuroscience Methods. 2004 ; Vol. 132, No. 1. pp. 101-107.
    @article{8e83c6a9e63f4cb08a00fb04baaec1d4,
    title = "The use of real-time PCR analysis in a gene expression study of Alzheimer's disease post-mortem brains",
    abstract = "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.",
    keywords = "β-Actin, Alzheimer's disease, GAPDH, Gene expression, Post-mortem brains, Real-time PCR",
    author = "Gutala, {Ramana V.} and Reddy, {P (Hemachandra)}",
    year = "2004",
    month = "1",
    day = "15",
    doi = "10.1016/j.jneumeth.2003.09.005",
    language = "English (US)",
    volume = "132",
    pages = "101--107",
    journal = "Journal of Neuroscience Methods",
    issn = "0165-0270",
    publisher = "Elsevier",
    number = "1",

    }

    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)

    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 - β-Actin

    KW - Alzheimer's disease

    KW - GAPDH

    KW - Gene expression

    KW - Post-mortem brains

    KW - Real-time PCR

    UR - http://www.scopus.com/inward/record.url?scp=0346995416&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=0346995416&partnerID=8YFLogxK

    U2 - 10.1016/j.jneumeth.2003.09.005

    DO - 10.1016/j.jneumeth.2003.09.005

    M3 - Article

    VL - 132

    SP - 101

    EP - 107

    JO - Journal of Neuroscience Methods

    JF - Journal of Neuroscience Methods

    SN - 0165-0270

    IS - 1

    ER -