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  • 1
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Proteinase inhibitor-DNA synthesis ; Mitotic activity ; Rat(Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous studies with rats have shown that a single oral dose of the proteinase inhibitor Camostate (FOY-305) induces release of cholecystokinin (CCK) into the circulation, which lasts for 3 to 6h. This transient endogenous release of hormone results in a depletion of pancreatic enzyme stores within 1 h and an increase in total rate of protein synthesis, which peaks at 6 to 9 h. At the level of individual enzyme biosynthesis a transient decrease in amylase and an increase in trypsinogen and chymotrypsinogen is observed. In the present study the time course of DNA synthesis and the labeling index of 5 populations of pancreatic cells have been analysed following a single oral dose of 50 or 100 mg/kg proteinase inhibitor, using in vivo labeling with 12 μCi/g body weight 3H-thymidine 1 h prior to sacrifice of the animals. DNA synthesis did not change during the initial 12 h following inhibitor feeding and then showed a phasic increase with a peak (20-fold) at 24h and intermediate increases (4- to 5-fold) at 18 and 36 h, respectively. From the 5 pancreatic cell populations studied by autoradiography the labeling indices of interlobular duct cells and islet cells did not change over the entire observation period. Acinar cells, intralobular duct cells and interstitial cells showed a marked increase in labeling index with peak values at 24h, which were 20-fold in acinar cells and 5.5- and 8.5-fold in intralobular duct cells and interstitial cells, respectively. The data demonstrate a significant growth response of pancreatic acinar tissue after a single episode of endogenous CCK-release, which is similar in extent, time course and cellular source as previously demonstrated during persistent stimulation of the pancreas by prolonged infusion of the CCK-analogue caerulein.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 113 (1971), S. 322-343 
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Human ; Release mechanism ; Cilia ; Junctional complex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung 1. An der exokrinen Pankreaszelle des Menschen lassen sich eine Basalzone (ER, Zellkern), eine Golgi-Zone und ein apikaler Zellpol unterscheiden. Die Zymogengranula entstehen durch Membranabknospung aus dem Golgi-Apparat und werden am apikalen Zellpol gespeichert. Die Lumenseite der exokrinen Zelle ist von dichtstehenden Mikrovilli besetzt (200–300 pro Zelloberfläche), die eine netzig-fibrilläre Längsstruktur zeigen. 2. Als morphologisches Substrat für den Vorgang der Extrusion werden wenige elektronendichte Granula am apikalen Pol beschrieben, die durch Aneinanderlagerung und Verschmelzung der Hüllmembranen Doppelformen von Granula oder kurze Granulaketten bilden. Die Extrusion wird abgeschlossen durch das Verschmelzen der Hüllmembran der einzelnen oder komplexen Granula mit der Plasmamembran, durch Ausbildung einer Öffnung in der Plasmamembran und durch das Ausfließen des Granuluminhaltes. Die Zelloberfläche von exokrinen Zellen mit lebhafter Extrusion zeigt nur spärlichen Besatz mit Mikrovilli. 3. Die exokrinen Zellen innerhalb eines Azinus werden an den apikalen und seitlichen Kontaktflächen durch ein differenziertes Schlußleistennetz (Zonulae occludentes, Z. adhaerentes, Desmosomen) verbunden, Tonofibrillen verlaufen bogenförmig von einer Verschlußzone zur nächsten und strahlen radiär ins Zytoplasma der exokrinen Zellen aus. 4. Die Epithelzellen der Schaltstücke sind durch ähnlich gebaute Haftzonen mit den exokrinen Zellen und untereinander verbunden; ihr Zytoplasma enthält einen kleinen Golgi-Apparat und regelmäßig langgestreckte Zilien (9+2-Muster), die aus einer trichterförmigen Vertiefung der Zelloberfläche entspringen und weit in das Schaltstücklumen hineinragen.
    Notes: Summary 1. The cytoplasm of human exocrine pancreatic cells may be divided into 3 regions: basal (ER, nucleus), Golgi zone and apical pole. The zymogen granules are condensed in the Golgi region and are stored at the apical pole. From the surface of a single exocrine cell 200 to 300 microvilli project into the lumen of the acinus, they contain an internal fibrillar structure oriented in the long-axis of each microvillus. 2. Before being released into the lumen of the acinus the content of zymogen granules becomes electron translucent and the limiting membranes of adjacent granules fuse to form duplex or short interconnected series of granules. The release is terminated by coalescence of the limiting membrane of single or complex granules with the plasmalemma, formation of an opening at the cell surface and extrusion of secretory material. Cells with signs of active release have only few microvilli on their luminal surface. 3. In a single acinus the apical and lateral surfaces of exocrine cells are connected by elaborate junctional complexes (zonulae occludentes, z. adhaerentes, desmosomes), in the adjacent cytoplasm curved bundles of tonofilaments run from one junctional complex to the other and extend also over a long distance into the cytoplasm. 4. The epithelial cells of the intercalated ducts are connected with the exocrine cells by the same pattern of junctional complexes and tonofilaments. Their cytoplasm contains a small Golgi complex and always a single cilium, which extends through a funnelshaped indentation of the cell surface over a long distance (6 to 8 μ) into the lumen of the intercalated duct.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 170 (1976), S. 203-219 
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Intracellular transport ; Cell fractionation ; Enzyme discharge
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The previous finding that intracellular transport of secretory proteins in the rat exocrine pancreas is accelerated by in vivo stimulation with a pancreatic secretagogue has been further analyzed. Using a radioassay for discharge of newly synthesized proteins, the rate of release was compared in control and prestimulated lobules. In control preparations discharge occurred with an initial lag period of 30 minutes and a maximum after two hours of incubation. After in vivo infusion of 5 × 10-8 g/hr. caerulein for 24 h in vitro discharge started after 10 minutes of in vitro incubation and attained a maximal rate after one hour. Using the same radioassay and several inhibitors of intracellular transport and granule discharge, it could be demonstrated that both processes were reduced to the same extent in controls and in lobules with accelerated transport. To obtain direct evidence for the degree of acceleration of the different transport steps between rough endoplasmic reticulum, Golgi complex and zymogen granules, the respective subcellular fractions of these organelles prepared and characterized ultrastructurally and biochemically. The rate of disappearance of newly formed proteins from rough microsomes and the appearance in smooth microsomes and zymogen granules were significantly increased after in vivo stimulation. The data substantiate an acceleration of the regular transport steps by the secretagogue. There was no indication that a high level of secretory activity leads to a rerouting of secretory proteins or to an omission of one of the regular steps in intracellular transport.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 175 (1976), S. 227-243 
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Golgi complex ; Glycoproteins ; Intracellular transport ; Secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using a double-label technique on isolated rat pancreatic lobules, the rate of synthesis and discharge of regular and fucosylated secretory proteins was studied under control conditions and after in vivo prestimulation with caerulein. Both labeled leucine and fucose were incorporated into pancreatic proteins at a linear rate, which was potentiated by in vivo stimulation. In pulse-chase experiments both regular and fucosylated secretory proteins were discharged into the medium in parallel. The in vivo pretreatment with caerulein caused an earlier discharge and increased the total amount released. Kinetic analysis of unstimulated (baseline) discharge of both classes of secretory proteins indicated a striking in vitro sensitivity by the previous in vivo treatment with caerulein. The biochemical data were compared to the fine structure of the Golgi complex under both control and prestimulated conditions. The Golgi stacks were composed of four to six individual cisternae which in some cases were connected by intercisternal pores. Transporting vesicles were observed fusing along the total length of the outermost cisterna on both the cis- and transside and with the lateral ends of the intermediate cisternae. Under control conditions only the last trans-cisterna contained some electron opaque material; in vivo prestimulation led to distension and filling of all cisternae in an individual Golgi-unit. Numerous stages of transformation of the last transcisterna into condensing vacuoles were observed, lending support to the hypothesis that during packaging of secretory products the membranes of the Golgi complex undergo a continuous turnover.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 193 (1978), S. 93-105 
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Cell fractionation ; Glycoproteins ; Membrane proteins ; Intracellular transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The transcellular movement of fucosylated glycoproteins has been studied in vitro using rat pancreatic lobules and cell fractionation procedures, and has been compared with the well established pathway of secretory proteins. Using tritiated leucine as pulse label for the latter, their translocation from the rough endoplasmatic reticulum into the Golgi complex and finally into zymogen granules could be followed. In the case of glycoproteins, 14C-fucose was incorporated mainly into the smooth microsomal fraction (representative of the Golgi complex) and only one third of this specific activity was transported into the zymogen granule fraction. A detailed analysis of this fraction after separation of the content of zymogen granules from their membranes revealed a predominant labeling of membrane glycoproteins by 14C-fucose. In comparison, leucine-labeled bulk proteins were found almost exclusively in the zymogen granule content fraction, with little radioactivity in the membrane fraction. The data indicate a concomitant synthesis of fucosylated glycoproteins destined in part for the zymogen granule membrane and to a greater amount associated with the smooth microsomal fraction. The results are discussed in the light of recent findings indicating that about 40% of the proteins in the zymogen granule membrane are made up of one major glycoprotein which could be involved in the mechanism of exocytosis.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 194 (1978), S. 447-462 
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Amino acid transport ; Hormone action ; Tight junction ; Gap junction ; Exocytosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Amino acid transport and incorporation have been studied in vitro in rat pancreatic lobules after maximal and supramaximal hormonal stimulation with caerulein. Incorporation into proteins was increased already after 30 and 120 min of maximal stimulation, but was decreased after the infusion of a supramaximal dose. Uptake of neutral amino acids was monitored using labeled leucine and α-aminoisobutyric acid (AIB). In the case of leucine the free pool was consistently reduced after maximal stimulation, while supramaximal doses led to an increase which could be potentiated by the addition of 2mM tetracaine. Using AIB, a significant increase in the intracellular pool was observed after maximal stimulation, conversely a decrease after supramaximal stimulation. Release of labeled leucine and AIB from preloaded lobules during incubation in the cold was significantly reduced after maximal secretory stimulation, but was found enhanced by 200 to 300 percent after supramaximal stimulation. No fine structural alterations at junctional complexes or at both the lateral and luminal plasma membranes were observed after maximal stimulation except an increased number of exocytotic figures at the luminal face. However, supramaximal stimulation led to progressive rarefaction of the tight junctional network and disintegration of the gap junctions. Concomitantly, an equal distribution of membrane particles on both faces of the plasma membrane together with a random occurrence of exocytotic figures were observed.
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  • 7
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Proteinase inhibitor ; Feedback regulation ; Cholecystokinin ; Fine structure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Application of a single dose of a new type of proteinase inhibitor camostate (FOY-305) via orogastric tube was used in rats to study the dose-response relationship of resulting pancreatic stimulation. Doses up to 10 mg/ kg failed to elicit any response, while significant decrease in enzyme content and increase in serum CCK-levels were observed with doses ranging from 25 to 400 mg/kg. A single dose of 100 mg/kg was selected for a time-sequence analysis, which revealed a 60 to 70% depletion of enzyme stores persisting over 6 h and reverting to control levels by 12 h. Peak increases in serum CCK-levels (15-fold above the elevation observed after regular food intake) were found after 30 min and persisted as an 8-to 10-fold elevation for at least 3 h, then declined to control levels by 9 h. This prolonged endogenous hormone release and resulting pancreatic stimulation were also verified in a separate group of animals in which volume, protein, and enzyme output were measured after cannulation of the pancreatic duct. While volume secretion was not altered by feeding a single dose of 100 mg/kg FOY-305, protein and enzyme output increased 2-to 3-fold over a period of 7 h. Fine-structural analysis of the pancreas demonstrated efficient depletion of zymogen granules from acinar cells with all doses between 50 and 400 mg/kg, accompanied by the appearance of membrane material in the acinar lumina at 3 and 6 h. The same transient increase in the number of lysosomal bodies predominantly containing mitochondria with all doses above 50 mg/kg was interpreted as increased organelle turnover due to persisting hormonal stimulation.
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  • 8
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Proteinase inhibitor ; Cholecystokinin ; Protein synthesis ; Enzyme synthesis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Oral application of a single dose of a new synthetic proteinase inhibitor Camostate (Foy-305) in male Wistar rats was carried out together with studies of in vitro amino acid incorporation followed by separation of proteins by two-dimensional gel electrophoresis. The aim of this experiment was to analyze changes produced by the inhibitor in total protein and individual enzyme biosynthesis. Administration of 100 mg/kg Foy-305 resulted in significant inhibition of total pancreatic protein synthesis, without changes in fractional rates for individual enzymes. 50 mg/kg Foy-305 induced a 10-fold elevation of cholecystokinin (CCK) levels in serum; this persisted for 3 h and led to a significant increase in the total rate of protein synthesis with peak values at 6 and 9 h (78% and 84% above control levels, respectively), returning to control by 15h. Changes in fractional rates of synthesis occurred with a latency of 6 h and were restricted to amylase and the anionic form of trypsinogen and chymotrypsinogen. Amylase biosynthesis decreased by about 40% from control levels at 9 h to return to control levels by 15 h. Increased synthesis of trypsinogen and chymotrypsinogen was observed; this was also phasic. The results show similar enzyme-specific regulation as previously described for exogenous CCK stimulation and for the adaptation of the pancreas to diets enriched in protein. They demonstrate the effectiveness of pulsatory endogenous hormone release in the regulation of protein synthesis.
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  • 9
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Degranulation ; In vivo stimulation ; Membrane degradation ; Protein synthesis ; I ntracellular transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Prolonged secretory stimulation of the exocrine pancreas in the rat by in vivo infusion of caerulein leads to a rapid degranulation of the organ associated with a progressive reduction in the size of the zymogen granules. During the first six to twelve hours of stimulation Golgi complexes are enlarged and several structural forms of multivesicular bodies are found indicating a lysosomal degradation of membrane material in the Golgi area. Maximum secretory activity is obtained after a 24 hour infusion, Golgi complexes appear fragmented, the secretory granules measure only 1/3 to 1/4 their normal size. Thereafter, in spite of a continuous stimulation, the exocrine cells regranulate progressively up to 72 hours of infusion. This regranulation is associated with massive enlargement of the Golgi complexes. The phasic adaptation of the exocrine pancreas to prolonged stimulation, concluded from the structural studies, was confirmed by biochemical analysis of protein synthesis, intracellular transport and enzyme discharge. Pancreatic protein synthesis as measured by the incorporation of tritiated leucine remained unchanged during the first six hours of stimulation, then increased reaching a maximum of 230% of the control levels after 24 hours of infusion. After 48 and 72 hours the rate of protein synthesis decreased again to normal values. Most pronounced changes were observed in the kinetics of intracellular transport of newly synthesized proteins. Using pulse-chase incubation of prestimulated pancreatic lobules, the rate of transition of secretory proteins through the cell increased consistently with prolonged infusion periods reaching maximal acceleration after 24 hours. Newly synthesized proteins were transported and segregated up to ten times faster than in controls. After a maximum at 24 hours transport returned to normal rates after 72 hours of infusion. Enzyme secretion, measured for amylase, followed a similar pattern of stimulation. The results suggest a phasic adaptation of the exocrine pancreatic cell to prolonged stimulation. They demonstrate for the first time the possibility of an acceleration of intracellular transport by means of secretagogues.
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  • 10
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Caerulein ; DNA synthesis ; Mitotic activity ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous studies from our laboratory indicate that the adaptive response of the exocrine pancreas of the rat to prolonged stimulation with optimal doses of caerulein (0.25 μg × kg-1 × h-1) follows a characteristic time course in which each step in the secretory pathway is activated. The immediate response is the depletion of zymogen-granule stores followed by coordinate and anticoordinate changes in individual rates of (pro-)enzyme synthesis after a lag period of 2 h. The sum of such changes leads to an increase in total rate of protein synthesis by 3 h which is combined with acceleration of intracellular transport packaging and granule discharge. In the present study the time course of DNA synthesis and the labeling index of five populations of pancreatic cells have been analyzed after caerulein stimulation for periods ranging from 6 to 72 h, using in vivo labeling with 1 μCi/g 3H-thymidine 1 h prior to sacrifice of the animals. DNA synthesis did not change during the initial 18 h in spite of persistent stimulation indicated by a 80% reduction of enzyme content. Following this lag period a sharp rise in DNA synthesis 20- to 25-fold above control levels was observed, which decreased by 48 h to reach control levels by 72 h. Increase in DNA synthesis was most pronounced in animals with lowest enzyme content in the pancreas. From the five cell populations studied by autoradiography interlobular duct cells and islet cells had no significant increase in labeling index at any time of stimulation. Acinar cells, intralobular duct cells and interstitial cells showed a marked increase in labeling index after a latent period of 18 h with peak values at 36 h 30 to 50 times higher in intralobular duct and acinar cells, respectively, and 4 times higher in interstitial cells. The increased labeling indices in all three cell populations reverted to lower values at 48 h and reached control values by 72 h. The data indicate a phasic and limited growth response of the rat exocrine pancreas to persistent stimulation with acinar cells as the major contributing cell population.
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