Quantitative real-time polymerase chain reaction measurement of regulators of G-protein signaling mRNA levels in mouse tissues

Deborah M. Kurrasch, Jie Huang, Thomas M. Wilkie, Joyce J. Repa

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

Regulators of G-protein signaling (RGS) play a critical role in G-protein-coupled receptor signaling in mammalian cells. RGS proteins are GTPase-accelerating proteins (GAPs) for α subunits of heterotrimeric G proteins of the G i and G q class. RGS GAPs can modulate the frequency and duration of G-protein signaling and may constitute a new family of therapeutic targets. Identifying the tissue distribution and cellular localization of RGS proteins has been hindered by the lack of effective antibodies for immunodetection. The measurement of mRNA levels for RGS proteins, however, can provide insight into their tissue specificity and regulation. This article describes the use of a highly sensitive and rapid method for measuring RGS mRNA in mouse tissues. This quantitative real-time polymerase chain reaction method is established for the 19 reported mouse RGS genes and is used to study the tissue distribution of the R4 family of RGS genes and the diurnal regulation of RGS16 in mouse liver.

Original languageEnglish (US)
Pages (from-to)3-15
Number of pages13
JournalMethods in Enzymology
Volume389
DOIs
StatePublished - 2004

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GTP-Binding Protein Regulators
Polymerase chain reaction
RGS Proteins
Real-Time Polymerase Chain Reaction
Tissue
Messenger RNA
GTP Phosphohydrolases
Tissue Distribution
Genes
Heterotrimeric GTP-Binding Proteins
Organ Specificity
Protein Subunits
G-Protein-Coupled Receptors
GTP-Binding Proteins
Liver
Cells
Antibodies
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

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title = "Quantitative real-time polymerase chain reaction measurement of regulators of G-protein signaling mRNA levels in mouse tissues",
abstract = "Regulators of G-protein signaling (RGS) play a critical role in G-protein-coupled receptor signaling in mammalian cells. RGS proteins are GTPase-accelerating proteins (GAPs) for α subunits of heterotrimeric G proteins of the G i and G q class. RGS GAPs can modulate the frequency and duration of G-protein signaling and may constitute a new family of therapeutic targets. Identifying the tissue distribution and cellular localization of RGS proteins has been hindered by the lack of effective antibodies for immunodetection. The measurement of mRNA levels for RGS proteins, however, can provide insight into their tissue specificity and regulation. This article describes the use of a highly sensitive and rapid method for measuring RGS mRNA in mouse tissues. This quantitative real-time polymerase chain reaction method is established for the 19 reported mouse RGS genes and is used to study the tissue distribution of the R4 family of RGS genes and the diurnal regulation of RGS16 in mouse liver.",
author = "Kurrasch, {Deborah M.} and Jie Huang and Wilkie, {Thomas M.} and Repa, {Joyce J.}",
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T1 - Quantitative real-time polymerase chain reaction measurement of regulators of G-protein signaling mRNA levels in mouse tissues

AU - Kurrasch, Deborah M.

AU - Huang, Jie

AU - Wilkie, Thomas M.

AU - Repa, Joyce J.

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