Affinity-purified, monospecific rabbit antibodies against rat pancreatic a-amylase and bovine pancreatic a-chymotrypsinogen were used for immunoferritin observations of ultrathin frozen sections of mildly fixed exocrine pancreatic tissue from secretion-stimulated (pilocarpine) rats and from overnight-fasted rats and guinea pigs. The labeling patterns for both antibodies were qualitatively alike : Labeling occurred in (a) the cisternae of the rough endoplasmic reticulum (RER) including the perinuclear cisterna, in (b) the peripheral area between the RER and cis-Golgi face, and (c) all Golgi cisternae, condensing vacuoles, and secretory granules .Labeling of cytoplasmic matrix was negligible . Structures that appeared to correspond to rigid lamellae were unlabeled . Differences in labeling intensities indicated that concentration of the zymogens starts at the boundary of the RER and cis-side of the Golgi complex . These data support the view that the Golgi cisternae are involved in protein processing in both stimulated and unstimulated cells and that Golgi cisternae and condensing vacuoles constitute a functional unit .KEY WORDS secretory proteins . Golgi complex . exocrine pancreas . immunoferritin cytochemistry . cryoultramicrotomyThe intracellular route that secretory proteins take during their migration from the membrane-bound polyribosomes towards the secretory granules has not yet been mapped in detail . A main problem is what role the Golgi complex plays in protein processing . The Golgi complex in the exocrine pancreatic cell, as in other secretory cells, is composed of a stack of cisternae with peripheral elements consisting of vesicles and tubules at its cis-(immature, proximal, convex, outer) side, and condensing vacuoles and small membranous struc-J . CELL BIOLOGY

Gastric surface mucous cells (SMC), mucous neck cells (MNC) and their undifferentiated and immature precursors were studied by light and electron microscopic histochemistry. The secretory granules of SMC were smaller, more electron dense and more reactive to PAS and its analogues than those of MNC. Alcian blue demonstrated that the mucus of SMC was acidic and that of MNC was neutral. The periodic acid-thiocarbohydrazide-silver proteinate method revealed the pressence of carbohydrates in the golgi apparatus, condensing vacuoles, secretory granules, apical vesicles and tubules and cell coat. Maturation of SMC during their migration towards the free surface was reflected by an increase in size and number of secretory granules, an increase of RER and microfilaments, and a decrease of microvilli and apical vesicles and tubules. The secretory granules of older SMC were less acidic and possessed a proteinaceous core. Most MNC were fully differentiated, but some immature MNC containing only a few granules were found. Furthermore, undifferentiated cells and intermediates between SMC and MNC were also observed. The presence of both transitional and intermediate forms indicates that both SMC, and MNC arise from the same population of undifferentiated cells. Incorporation of 3H-thymidine revealed that undifferentiated cells, use isthmic SMC, MNC and intermediate cells are proliferative. No proliferative activity was found in foveolar SMC, parietal, chief, fibrillovesicular or endocrine cells.

The distal wall of the groove between the rat forestomach and glandular stomach is lined with a special type of columnar cells (CCGG) and with fibrillovesicular cells (FVC). The cardiac glands contain cardiac mucosa (CMC) and serous cells (CSC). The CCGG contain small mucous granules and special vesicles and tubules. The CMC are filled with large mucous granules and resemble mucous neck cells. The CSC are filled with large proteinaceous granules. The FVC are characterized by long microvilli, apical bundles of microfilaments and a complex "tubulovesicular system". The pattern of 3H-thymidine incorporation and the presence of immature and transitional forms indicate a possible origin of all the cell types concerned from a common undifferentiated precursor. The membranes of the tubulovesicular system of FVC as well as the apical cell membrane were reactive to Thiéry's carbohydrate stain. However, lanthanum tracing of the extracellular space and ultrastructural stereoscopy did not reveal a permanent continuity between both membrane systems. The absence of 3H-thymidine label showed that FVC were not proliferative. The structural characteristics of FVC do not account for a secretory, resorptive or receptive function. The special arrangement of microfilaments and the tubulovesicular system suggests an ability to fast changes in surface area.

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Frog pancreatic tissue was pulse-labelled in vitro with 3H-leucine and protein transport was studied in exocrine cells by electron microscope autoradiography. The proteins appeared to be synthesized in the RER and transported to the secretory granules along a similar route and with the same velocity as previously described under in vitro conditions. Evidence was obtained for the involvement of the vesicular and tubular elements at the periphery of the Golgi system in transferring protein from the RER to the Golgi cisternae. Kinetics of the release of newly synthesized proteins from the RER and their appearance in the condensing vacuoles are discussed and related to results reported from other tissues. The transport velocity in this poikilothermic system was studied in relation to the incubation temperature and compared with results reported from its mammalian counterpart. At temperatures between 20 and 30 degrees C intracellular protein transport occurs faster in the frog than in the Guinea pig pancreas. At higher temperature the transport process was severely disturbed in the frog.

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Pancreatic secretion in the rat was stimulated in vivo by pilocarpine injection causing 90% of the storage granules to be discharged within 2 h . Incubation in vitro with [14C]sorbitol indicated that maximal ingestion of this extracellular space marker occurred 3 h after secretogogue injection . Morphological cell membrane measurements on cells with stimulated secretion revealed a simultaneous decrease in amount of membrane bordering the microvilli at the cell apex, lamellar processes, and infoldings present at the latero-basal face of these cells. In 3-h stimulated cells, having the average zymogen granule content characteristic for that phase of secretion, ferritin treatment in vitro showed that the infoldings and related fragmentation vesicles had ingested ferritin and could consequently be considered as being transport vehicles for redundant cell membrane . During stimulated secretion numerous vesicles and vacuoles appeared in the apical cytoplasm . Part of these structures were postulated to be related to the Golgi complex and were discussed in relation to secretory protein transport . Another part of these structures was assumed to have an endocytotic nature, although they never contained ferritin .