A fast carrier chromatin immunoprecipitation method applicable to microdissected tissue samples

Haiping Hao; Hester Liu; Gregory Gonye; James S. Schwaber

doi:10.1016/j.jneumeth.2008.04.002

A fast carrier chromatin immunoprecipitation method applicable to microdissected tissue samples

Haiping Hao, Hester Liu, Gregory Gonye, James S. Schwaber

Research output: Contribution to journal › Article › peer-review

Abstract

Transcriptional regulation studies of CNS neurons are complicated by both cellular diversity and plasticity. Microdissection of specific functionally related populations of neurons can greatly reduce these issues, but typically excludes the use of many technologies due to tissue requirements, such as Chromatin Immunoprecipitation (ChIP), a powerful tool for studying in vivo protein-DNA interactions. We have developed a fast carrier ChIP (Fast CChIP) method for analyzing specific in vivo transcription factor-DNA interactions in as little as 0.2 mm³ brain tissue. Using an antibody against phosphorylated cyclic-AMP response element binding (CREB) protein, we confirmed phospho-CREB (pCREB) binding at the c-fos gene promoter. Then we further demonstrated the applicability of Fast CChIP in determining hypertension-induced pCREB binding at the c-fos gene promoter in the rat nucleus tractus solitarius (NTS), confirming CREB's role in mediating hypertension-induced c-fos expression. This method will be broadly applicable to individual brain nucleus and biopsy/surgical samples.

Original language	English (US)
Pages (from-to)	38-42
Number of pages	5
Journal	Journal of Neuroscience Methods
Volume	172
Issue number	1
DOIs	https://doi.org/10.1016/j.jneumeth.2008.04.002
State	Published - Jul 15 2008
Externally published	Yes

Keywords

CNS
ChIP
Chromatin immunoprecipitation
Fast CChIP
Microdissection
Transcription regulation
qPCR

ASJC Scopus subject areas

General Neuroscience

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10.1016/j.jneumeth.2008.04.002

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title = "A fast carrier chromatin immunoprecipitation method applicable to microdissected tissue samples",

abstract = "Transcriptional regulation studies of CNS neurons are complicated by both cellular diversity and plasticity. Microdissection of specific functionally related populations of neurons can greatly reduce these issues, but typically excludes the use of many technologies due to tissue requirements, such as Chromatin Immunoprecipitation (ChIP), a powerful tool for studying in vivo protein-DNA interactions. We have developed a fast carrier ChIP (Fast CChIP) method for analyzing specific in vivo transcription factor-DNA interactions in as little as 0.2 mm3 brain tissue. Using an antibody against phosphorylated cyclic-AMP response element binding (CREB) protein, we confirmed phospho-CREB (pCREB) binding at the c-fos gene promoter. Then we further demonstrated the applicability of Fast CChIP in determining hypertension-induced pCREB binding at the c-fos gene promoter in the rat nucleus tractus solitarius (NTS), confirming CREB's role in mediating hypertension-induced c-fos expression. This method will be broadly applicable to individual brain nucleus and biopsy/surgical samples.",

keywords = "CNS, ChIP, Chromatin immunoprecipitation, Fast CChIP, Microdissection, Transcription regulation, qPCR",

author = "Haiping Hao and Hester Liu and Gregory Gonye and Schwaber, {James S.}",

year = "2008",

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doi = "10.1016/j.jneumeth.2008.04.002",

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T1 - A fast carrier chromatin immunoprecipitation method applicable to microdissected tissue samples

AU - Hao, Haiping

AU - Liu, Hester

AU - Gonye, Gregory

AU - Schwaber, James S.

PY - 2008/7/15

Y1 - 2008/7/15

N2 - Transcriptional regulation studies of CNS neurons are complicated by both cellular diversity and plasticity. Microdissection of specific functionally related populations of neurons can greatly reduce these issues, but typically excludes the use of many technologies due to tissue requirements, such as Chromatin Immunoprecipitation (ChIP), a powerful tool for studying in vivo protein-DNA interactions. We have developed a fast carrier ChIP (Fast CChIP) method for analyzing specific in vivo transcription factor-DNA interactions in as little as 0.2 mm3 brain tissue. Using an antibody against phosphorylated cyclic-AMP response element binding (CREB) protein, we confirmed phospho-CREB (pCREB) binding at the c-fos gene promoter. Then we further demonstrated the applicability of Fast CChIP in determining hypertension-induced pCREB binding at the c-fos gene promoter in the rat nucleus tractus solitarius (NTS), confirming CREB's role in mediating hypertension-induced c-fos expression. This method will be broadly applicable to individual brain nucleus and biopsy/surgical samples.

AB - Transcriptional regulation studies of CNS neurons are complicated by both cellular diversity and plasticity. Microdissection of specific functionally related populations of neurons can greatly reduce these issues, but typically excludes the use of many technologies due to tissue requirements, such as Chromatin Immunoprecipitation (ChIP), a powerful tool for studying in vivo protein-DNA interactions. We have developed a fast carrier ChIP (Fast CChIP) method for analyzing specific in vivo transcription factor-DNA interactions in as little as 0.2 mm3 brain tissue. Using an antibody against phosphorylated cyclic-AMP response element binding (CREB) protein, we confirmed phospho-CREB (pCREB) binding at the c-fos gene promoter. Then we further demonstrated the applicability of Fast CChIP in determining hypertension-induced pCREB binding at the c-fos gene promoter in the rat nucleus tractus solitarius (NTS), confirming CREB's role in mediating hypertension-induced c-fos expression. This method will be broadly applicable to individual brain nucleus and biopsy/surgical samples.

KW - CNS

KW - ChIP

KW - Chromatin immunoprecipitation

KW - Fast CChIP

KW - Microdissection

KW - Transcription regulation

KW - qPCR

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A fast carrier chromatin immunoprecipitation method applicable to microdissected tissue samples

Abstract

Keywords

ASJC Scopus subject areas

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