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Journal of Neuroscience, Vol 15, 7966-7978, Copyright © 1995 by Society for Neuroscience
Differential regulation of neuronal nicotinic ACh receptor subunit genes in cultured neonatal rat sympathetic neurons: specific induction of alpha 7 by membrane depolarization through a Ca2+/calmodulin- dependent kinase pathway
P De Koninck and E Cooper
Department of Physiology, McGill University, Montreal, Quebec, Canada.
We have examined the regulation of neuronal nicotinic ACh receptor (nAChR)
genes and ACh-evoked currents by neonatal rat sympathetic neurons
developing in culture. These neurons contain 5 nAChR transcripts: alpha 3,
alpha 5, alpha 7, beta 2, and beta 4. When developing in culture, the
neurons express 4 of these transcripts, alpha 3, alpha 5, beta 2, and beta
4, at levels similar to those in neurons developing in vivo: alpha 3 mRNA
levels increase two- to threefold over the first week, whereas the levels
for alpha 5, beta 2, and beta 4 remain essentially constant. In contrast,
alpha 7 mRNA levels drop by 60-75% within the first 48 hr and remain low.
We show that during the first week, the ACh-evoked current densities on
these cultured neurons increase twofold and correlate well with the
increase in alpha 3 mRNA levels. Depolarizing the neurons with 40 mM KCl
for 1-2 d upregulates the alpha 7 gene; this specific change in alpha 7
mRNA level correlates with an increase in alpha-bungarotoxin (alpha-BTX)
binding on the surface of the neurons. Depolarization has little effect on
the expression of the other four transcripts, or on the magnitude or
kinetics of the ACh-evoked currents. Furthermore, activators or inhibitors
of protein kinase A (PKA), protein kinase C (PKC), or tyrosine kinase do
not affect nAChR transcript levels in these cultured neurons. The effect of
membrane depolarization on alpha 7 expression is a result of Ca2+ influx
through L-type Ca2+ channels, and we show that alpha 7 is upregulated
through a Ca2+/calmodulin-dependent protein kinase (CaM kinase) pathway.
The identification of CaM kinase as a link between activity and
neurotransmitter receptor expression may indicate a novel mechanism that
underlies some forms of synaptic plasticity.
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