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Non-neuronal cell-conditioned medium regulates muscarinic receptor expression in cultured sympathetic neurons

KE Smith and JA Kessler
Journal of Neuroscience 1 July 1988, 8 (7) 2406-2413; DOI: https://doi.org/10.1523/JNEUROSCI.08-07-02406.1988
KE Smith
Albert Einstein College of Medicine, Department of Neurology, Bronx, New York 10461.
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JA Kessler
Albert Einstein College of Medicine, Department of Neurology, Bronx, New York 10461.
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Abstract

Regulation of muscarinic receptor expression was examined in cultured sympathetic neurons of the neonatal rat superior cervical ganglion. Receptor concentration was determined by measuring binding of the muscarinic antagonist 1-quinuclidinyl[phenyl-4-3H] benzilate (3H-QNB). 3H-QNB bound to one apparent class of noninteracting sites on sympathetic neuron membranes with a Kd of 28.9 pM and a Bmax of 2.91 pmol/mg protein. Binding increased as a linear function of tissue protein and was saturable. The number of receptors per milligram protein increased approximately 6-fold during 16 d of culture, and receptor numbers were down-regulated by treatment with the agonist carbachol. These observations suggested that measurement of 3H-QNB binding would provide a reliable estimate of muscarinic receptor number on cultured sympathetic neurons. To determine whether nonneuronal cells produce soluble factors that influence muscarinic receptor expression, the effects of treatment with rat fibroblast-conditioned medium (RFCM) were examined. Exposure of sympathetic neurons to 50% RFCM resulted in a 57% decrease in muscarinic receptor numbers without a change in the apparent Kd. The decrease in 3H-QNB binding in response to RFCM was dose-related, with a minimum dose of 15% RFCM required to observe a significant effect. In contrast to the carbachol-induced down- regulation, the reduction in binding after RFCM treatment was not prevented by atropine, indicating that the effect was not mediated by acetylcholine stimulation of muscarinic receptors. Binding of 125I- alpha-bungarotoxin, which labels a nonfunctional membrane site distinct from nicotinic receptors, was not altered by treatment with RFCM, indicating the selectivity of the change in membrane muscarinic receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

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The Journal of Neuroscience: 8 (7)
Journal of Neuroscience
Vol. 8, Issue 7
1 Jul 1988
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Non-neuronal cell-conditioned medium regulates muscarinic receptor expression in cultured sympathetic neurons
KE Smith, JA Kessler
Journal of Neuroscience 1 July 1988, 8 (7) 2406-2413; DOI: 10.1523/JNEUROSCI.08-07-02406.1988

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Non-neuronal cell-conditioned medium regulates muscarinic receptor expression in cultured sympathetic neurons
KE Smith, JA Kessler
Journal of Neuroscience 1 July 1988, 8 (7) 2406-2413; DOI: 10.1523/JNEUROSCI.08-07-02406.1988
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