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Journal of Neuroscience, Vol 14, 5417-5428, Copyright © 1994 by Society for Neuroscience
Distinctive patterns of GABAA receptor subunit mRNAs in 13 cell lines
RF Tyndale, TG Hales, RW Olsen and AJ Tobin
Department of Pharmacology, School of Medicine, University of California, Los Angeles 90024.
We have investigated the GABAA receptor mRNA composition in 13 cell lines,
using 13 subunit-specific oligo-primers (alpha 1-6, beta 1-3, gamma 1-3,
and delta) and reverse transcriptase PCR amplification. Cell lines (B35,
B65, B103, B104, RINm5F, Rat1, PC12, C6, C17, C27, beta TC3, NB41A3,
AtT-20), derived from diverse tissue origins, were investigated in order to
identify homogeneous cellular sources with distinctive GABAA receptor
subunits. Fifteen GABAA receptor subunits have been cloned from mammalian
tissue (those listed above plus the retinal subunits rho 1 and rho 2). This
multiplicity of GABAA receptor subunits underlies the diverse pharmacology
of the GABAA receptor. Attempts to understand the regulation and
pharmacology of individual subunits and of the heterooligomeric receptor
combinations have been impeded by a lack of pure populations of cells
expressing GABAA receptor subunits. Permanent cell lines provide such a
resource. Each GABAA receptor subunit mRNA, alpha 1-5, beta 1-3, gamma 1-3,
and delta, was detected in at least one cell line. All cell lines examined
contained detectable levels of at least one GABAA receptor subunit mRNA.
Each cell line contained distinctive combinations of subunit mRNAs. None of
the cell lines examined contained detectable amounts of alpha 6 mRNA. These
cell lines, which transcribe GABAA receptor subunit mRNAs, provide useful
cellular sources for transcriptional and pharmacological studies. Our data
also suggest that endogenous GABAA receptor subunit mRNAs may be present in
cells that are routinely used for transfection studies, and that this
expression might confound interpretation of the studies. In the following
companion article, we have looked for functional GABAA receptor Cl- ion
channels in these cell lines, using the patch-clamp technique (Hales and
Tyndale, 1994).
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