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Am J Physiol Cell Physiol 289: C1379-C1387, 2005. First published July 20, 2005; doi:10.1152/ajpcell.00193.2005
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MUSCLE CELL BIOLOGY AND CELL MOTILITY

Smooth muscle adherens junctions associated proteins are stable at the cell periphery during relaxation and activation

Thomas J. Eddinger,1 Jessen D. Schiebout,1 and Darl R. Swartz2

1Biological Sciences, Marquette University, Milwaukee, Wisconsin; and 2Animal Sciences, Purdue University, West Lafayette, Indiana

Submitted 25 April 2005 ; accepted in final form 29 June 2005

This study was performed to determine the stability of the adherens junction (AJ)-associated proteins at the smooth muscle cell (SMC) plasma membrane during relaxing and activating conditions. Dog stomach, ileum, colon, and trachea tissues were stored in Ca2+-free PSS or regular PSS or were activated in 10 µM carbachol in PSS before rapid freezing. The tissues were subsequently sectioned and immunoreacted using antibodies for vinculin, talin, fibronectin, and caveolin to determine their cellular distribution in these tissues under these conditions. In all four tissues and under all three conditions, the distribution of these four proteins remained localized to the periphery of the cell. In transverse tissue sections, the AJ-associated proteins formed a distinct punctate pattern around the periphery of the SMCs at the plasma membrane. These domains alternated with the caveolae (as identified by the presence of caveolin). In longitudinal tissue sections, the AJ-associated proteins formed continuous tracks or staves, while the caveolae remained punctate in this dimension as well. Caveolin is not present in the tapered ends of the SMCs, where the AJ-associated proteins appear continuous around the periphery. Densitometry of the fluorophore distribution of these proteins showed no shift in their localization from the SMC periphery when the tissues were relaxed or when they were activated before freezing. These results suggest that under physiologically relaxing and activating conditions, AJ-associated proteins remain stably localized at the plasma membrane.

vinculin; talin; fibronectin; caveolin; stomach; ileum; colon; trachea



Address for reprint requests and other correspondence: T. J. Eddinger, Biological Sciences, Marquette Univ., 530 North 15th St., Milwaukee, WI 53233 (e-mail: thomas.eddinger{at}marquette.edu)




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