Am J Physiol Cell Physiol AJP: Regulatory, Integrative and Comparative Physiology
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Am J Physiol Cell Physiol 277: C1220-C1228, 1999;
0363-6143/99 $5.00
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Vol. 277, Issue 6, C1220-C1228, December 1999

EDITORIAL FOCUS
Asymmetric distribution of muscarinic acetylcholine receptors in Madin-Darby canine kidney cells

Laurie S. Nadler, Geetha Kumar, Thomas R. Hinds, Jacques C. Migeon, and Neil M. Nathanson

Department of Pharmacology, University of Washington School of Medicine, Seattle, Washington 98195-7750

We have characterized the muscarinic ACh receptors (mAChRs) expressed in Madin- Darby canine kidney (MDCK) strain II epithelial cells. Binding studies with the membrane-impermeable antagonist N-[3H]methylscopolamine demonstrated that mAChRs are ~2.5 times more abundant on the basolateral than on the apical surface. Apical, but not basolateral, mAChRs inhibited forskolin-stimulated adenylyl cyclase activity in response to the agonist carbachol. Neither apical nor basolateral mAChRs exhibited detectable carbachol-stimulated phospholipase C activity. Carbachol application to the apical or the basolateral membrane resulted in a threefold increase in intracellular Ca2+ concentration, which was completely inhibited by pertussis toxin on the apical side and partially inhibited on the basolateral side. RT-PCR analysis showed that MDCK cells express the M4 and M5 receptor mRNAs. These data suggest that M4 receptors reside on the apical and basolateral membranes of polarized MDCK strain II cells and that the M5 receptor may reside in the basolateral membrane of a subset of cells.

muscarinic receptor; cell polarity; adenosine triphosphate


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L. S. Nadler, G. Kumar, and N. M. Nathanson
Identification of a Basolateral Sorting Signal for the M3 Muscarinic Acetylcholine Receptor in Madin-Darby Canine Kidney Cells
J. Biol. Chem., March 23, 2001; 276(13): 10539 - 10547.
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