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The Journal of Cell Biology
The Rockefeller University Press
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DIRECT OBSERVATION OF TOAD BLADDER RESPONSE TO VASOPRESSIN JARED GRANTHAM, F. E. CUPPAGE, and DARRELL FANESTIL . From the Departments of Medicine and Pathology, University of Kansas Medical Center, Kansas City, Kansas 66103 It has been suggested that the lateral intercellular space is a pathway for transepithelial water flow in several tissues. Fundamental to this postulate is the observation that coincident with accelerated net fluid transport, either coupled to active electrolyte transport as in the gallbladder (1, 2), or due to passive water flow along osmotic gradients as in the collecting tubule (3, 4) and anuran membranes (5, 6), the lateral intercellular spaces widen . To correlate morphologic changes with fluid transport the epithelium is usually fixed with aldehydes or osmium tetroxide in preparation for electron microscopy. There has been no uniformity in the manner of applying the different fixative solutions to the different tissues . Water translocations may occur during the period of primary fixation, re- sulting in distortions of the lateral intercellular spaces unrelated to the physiologic function of the viable tissue (7) . The problem of fixation artifacts, specifically with regard to changes in the size of the intercellular channels, can be obviated by ob- serving the effect of hormone on the living tissue directly (3, 4) . In the present report we describe a simple method to observe directly the effect of ac- celerated hydrosmotic flow on the size of inter- cellular spaces in the isolated bladder of the toad, Bufo marinur. METHODS AND DISCUSSION Both lobes of a toad's urinary bladder were re- moved and washed in isotonic toad Ringer's solution. The composition of the medium is de- scribed elsewhere (8) . To reduce muscular con- traction during photography, atropine sulfate (0.05 mg/100 ml) was added to the serosal medium. In two studies atropine was not used and, although photographs could not be tak

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