Intracellular protein degradation and autophagy in isolated pancreatic acini of the rat.
Simultaneous investigation of protein degradation and autophagy of isolated exocrine pancreatic cells is carried out here for the first time in a systematic way by a complex biochemical, morphological and morphometrical approach. Protein degradation proceeds with a decreasing rate of 4-1.5 per cent per h over a 4-h period indicating a comparatively low degradation capacity. Cells in freshly isolated acini do not contain autophagic vacuoles but the latter appear within an hour in vitro and their quantity remains close to a steady state during the subsequent 3 h. Both traditional inhibitors of the autophagic-lysosomal pathway, e.g. vinblastine, leupeptin, and lysosomotropic amines together with the recently introduced 3-methyladenine, inhibit degradation to a similar maximal extent, offering the possibility of the estimation of the ratio of lysosomal/non-lysosomal degradation. In pancreatic acinar cells autophagic sequestration is unaffected and protein degradation is inhibited inside secondary lysosomes by leupeptin and lysosomotropic amines, while 3-methyladenine prevents the formation of autophagosomes. Vinblastine seems to act by inhibiting the fusion of autophagosomes with lysosomes and there is no evidence for the stimulation of autophagic sequestration by vinblastine in the present system. The effect of inhibitors of protein breakdown on protein synthesis is variable and does not correlate with their influence on degradation. Amino acids strongly stimulate protein synthesis, but in contrast to what is found in liver cells, they do not seem to affect protein degradation or autophagy significantly, thus indicating major regulatory differences of these processes between pancreatic acinar cells and hepatocytes.
Telbisz
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Department of General Zoology, Eotvos Lorand University, Budapest, Hungary.
Kovacs
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D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000588:Amines / Q000494:pharmacology; D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001343:Autophagy / Q000187:drug effects*; D002478:Cells, Cultured; D004791:Enzyme Inhibitors; D007700:Kinetics; D007976:Leupeptins / Q000494:pharmacology; D008247:Lysosomes / Q000201:enzymology / Q000378:metabolism; D008297:Male; D008854:Microscopy, Electron; D010179:Pancreas / Q000187:drug effects* / Q000648:ultrastructure; D014176:Protein Biosynthesis; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D020318:Rats, Long-Evans; D014747:Vinblastine / Q000494:pharmacology
D000588:Amines; D000596:Amino Acids; D004791:Enzyme Inhibitors; D007976:Leupeptins; D011506:Proteins; C025946:3-methyladenine; D014747:Vinblastine; C032854:leupeptin; D000225:Adenine