[2] Pancreatic lobules in the in vitro study of pancreatic acinar cell function (original) (raw)

Methods in Enzymology, 1983

Abstract

Publisher Summary This chapter discusses the methods for the preparation and study of pancreatic lobules. When properly prepared, pancreatic lobules and dissociated acini, show similar in vitro responses to varying levels of exogenously added secretagogues. The use of pancreatic lobules has two advantages over the use of dispersed acini: (1) the ease of preparation and (2) the omission of degradative enzymes and chelators during tissue preparation. The use of dispersed acini shows two advantages over the use of lobules: (1) the increased access of labeled probes to the plasma membrane, and (2) the apparent removal of nerve endings from isolated acini. Purified preparations of secretory proteins can be extracted from a crude fraction of zymogen granules or obtained physiologically after carbamylcholine-induced secretion of proteins into the incubation medium. The high resolution achieved in the separation of exocrine pancreatic proteins by two-dimensional isoelectric focusing/sodium dodecyl sulfate (IEF/SDS)-gel electrophoresis allows the precise measurement of biosynthetic rates of individual secretory products.

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