A cytochemical study on the pancreas of the guinea pig. II. Functional variations in the enzymatic activity of microsomes

Abstract

Microsomes were isolated from the pancreas of starved and fed guinea pigs. In the first case, the gland was removed from animals starved for 48 hours; in the second, the pancreas was excised 1 hour after the beginning of a meal that ended a fast of 48 hours. These are referred to below as fed animals. In both cases the tissue was homogenized in 0.88 M sucrose and the microsomes obtained by centrifuging the mitochondrial supernatant at 105,000 g for 60 minutes. In starved animals the content of the endoplasmic reticulum of the exocrine cells and the content of the microsomes were found to be of low or moderate density. In fed guinea pigs the cavities of the reticulum frequently contained dense intracisternal granules and the microsomes were distinguished by a content of high density sometimes in the form of recognizable intracisternal granules. In starved animals, the microsomes were found to account for 5 to 20 per cent of the trypsin-activatable proteolytic activity and ribonuclease activity of the whole cell, whereas in fed animals they contained uniformly almost 30 per cent of these activities. In fed animals the dense, cohesive content of the microsomes (intracisternal granules) could be isolated by breaking up the microsomes with dilute (0.1 per cent) deoxycholate solutions and separating microsomal subfractions by differential centrifugation. The specific enzymatic activities of a heavy microsomal subfraction rich in intracisternal granules were almost equal to those of isolated purified zymogen granules. The ribonucleoprotein particles attached to the microsomal membranes could be isolated by the same technique and found also to exhibit some of the same enzymatic activities. Corresponding subfractions isolated from the microsomes of starved animals were considerably less active. The relevance of these findings for the synthesis and intracellular transport of protein in the exocrine cell of the pancreas is discussed.