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Am J Physiol Cell Physiol 280: C155-C165, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 1, C155-C165, January 2001

A proposed mechanism for the potentiation of cAMP- mediated acid secretion by carbachol

Yuko Muto1, Taku Nagao1, Maki Yamada2,3, Katsuhiko Mikoshiba2,3, and Tetsuro Urushidani1

1 Laboratory of Pharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033; 2 Department of Molecular Neurobiology, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639; and 3 Laboratory for Developmental Neurobiology, Brain Science Institute, Institute of Physical and Chemical Research, Wako, Saitama 351-0198, Japan

Acid secretion in isolated rabbit gastric glands was monitored by the accumulation of [14C]aminopyrine. Stimulation of the glands with carbachol synergistically augmented the response to dibutyryl cAMP. The augmentation persisted even after carbachol was washed out and was resistant to chelated extracellular Ca2+ and to inhibitors of either protein kinase C or calmodulin kinase II. Cytochalasin D at 10 µM preferentially blocked the secretory effect of carbachol and its synergism with cAMP, whereas it had no effect on histamine- or cAMP-stimulated acid secretion within 15 min. Cytochalasin D inhibited the carbachol-stimulated intracellular Ca2+ concentration ([Ca2+]i) increase due to release from the Ca2+ store. Treatment of the glands with cytochalasin D redistributed type 3 inositol 1,4,5-trisphosphate receptor (the major subtype in the parietal cell) from the fraction containing membranes of large size to the microsomal fraction, suggesting a dissociation of the store from the plasma membrane. These findings suggest that intracellular Ca2+ release by cholinergic stimulation is critical for determining synergism with cAMP in parietal cell activation and that functional coupling between the Ca2+ store and the receptor is maintained by actin microfilaments.

inositol 1,4,5-trisphosphate receptor; cytoskeleton; cytochalasin D; calcium; parietal cell; rabbit


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