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Articles in PresS, published online ahead of print August 8, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00410.2002
Submitted on May 15, 2002
Accepted on August 1, 2002
1 Gazes Cardiac Research Institute, Medical University of South Carolina and the Department of Veterans Affairs Medical Center Charleston, Charleston, SC, USA
* To whom correspondence should be addressed. E-mail: cooperge{at}musc.edu.
One mechanism for the reappearance of G protein-coupled receptors after agonist activation is microtubule-based transport. In pressure-overload cardiac hypertrophy there is down-regulation of G protein-coupled receptors and the appearance of a densified microtubule network extensively decorated by a microtubule-associated protein, MAP 4. Our hypothesis is that overdecoration of a dense microtubule network with this structural protein, as in hypertrophied myocardium, would impede receptor recovery. We tested this hypothesis by studying muscarinic acetylcholine receptor internalization and recovery after agonist stimulation in neuroblastoma cells. Exposure of cells to carbachol, a muscarinic receptor agonist, decreased membrane receptor binding activity. After carbachol withdrawal, receptor binding recovered towards the initial value. When microtubules were depolymerized before carbachol withdrawal, mAChR recovery was only 44% of that in intact cells. Cells were then infected with an adenovirus containing MAP 4 cDNA. MAP 4 protein decorated the microtubules extensively, and receptor recovery upon carbachol withdrawal was reduced to 54% of control. Thus, muscarinic receptor recovery after agonist exposure is microtubule dependent, and MAP 4 decoration of microtubules inhibits receptor recovery.
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