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Journal of Neuroscience, Vol 13, 2662-2671, Copyright © 1993 by Society for Neuroscience
Coexpression of multiple acetylcholine receptor genes in neurons: quantification of transcripts during development
RA Corriveau and DK Berg
Department of Biology, University of California, San Diego, La Jolla 92093-0322.
A large family of genes encoding subunits of nicotinic ACh receptors
(AChRs) has been identified in vertebrates and shown to be expressed in the
nervous system. The multiplicity of genes raises questions about which gene
products coassemble to produce native receptor subtypes and how the
expression of receptor genes is regulated in neurons. We report here that
five neuronal AChR genes are expressed in the chick ciliary ganglion at
both early and late times in development. Quantitative RNase protection
experiments demonstrated that at embryonic day 18 (E18) the ganglion
contains about 1800 copies of alpha 7 transcript per neuron, 900 copies of
alpha 3 transcript per neuron, and 200-300 copies each of alpha 5, beta 2,
and beta 4 transcripts per neuron. The same five genes are expressed at
significantly lower levels at E8 but show the same rank order of abundance
in transcripts per neuron. Few, if any, transcripts were found for the
alpha 2, alpha 4, alpha 8, and beta 3 AChR genes in ciliary ganglion RNA at
either E8 or E18. The 6- and 13- fold increases previously reported for two
classes of AChRs on the neurons between E8 and E18 approximate the
4-14-fold increases observed here in AChR gene mRNA levels per neuron over
the same time period. The alpha 3, alpha 5, alpha 7, and beta 4 genes have
previously been correlated with subunits of ciliary ganglion AChRs, but the
beta 2 gene has not. The abundance of beta 2 transcripts raises the
possibility either that the known AChRs in the ganglion have a more complex
subunit composition than previously described or that additional receptor
subtypes remain to be discovered. Northern blot analysis revealed no
changes in transcript pattern for the alpha 3, alpha 5, and beta 4 genes
between E8 and E18; a small change may occur in the transcript pattern for
the alpha 7 gene. In situ hybridizations demonstrated that alpha 5 and beta
4 transcripts are expressed in essentially all ciliary ganglion neurons as
has been shown previously for the more abundant alpha 3 transcript and
inferred for the alpha 7 transcript. The results indicate that neurons can
stably coexpress multiple AChR genes, including three of the alpha type,
and that transcript levels may be rate limiting for accumulation of AChRs
during development.
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