Stimulated secretion of lysosomal enzymes by cells in culture.
F9 mouse teratocarcinoma and PyS-2 cells in culture incubated with monovalent cations in buffered sucrose solution (0.25 M) can secrete as much as 40% of their total lysosomal enzymes into the medium within 30 min. Longer incubation does not lead to further loss of enzyme, suggesting that only a certain fraction of lysosomes is capable of discharge. The simultaneous presence of sucrose and cation, each at the respective optimal concentrations of 0.25 and 0.15 M, is required for lysosomal discharge (i.e. twice isoosmolarity). The cells remain fully viable. Sodium ions are more effective than lithium and potassium ions, whereas amines and divalent cations are less effective. Other sugars including glucose can replace sucrose to varying extents. Secretion is accompanied by a rapid short-lived rise in the level of cAMP. Forskolin as well as agents that activate G protein such as cholera toxin, AlF4-, and vanadate ions also increase the rate of secretion. Sucrose-Na+ stimulation takes place independently of changes in influx or efflux of calcium ions or changes in the levels of extracellular or free intracellular calcium ions. Neomycin, an inhibitor of phospholipase C, has little effect on secretion. Our results suggest that the secretion observed is mediated by a cAMP-dependent mechanism involving G proteins. Calcium ions and phospholipase C appear to play little or no part in the activation process.
Warren
L
L
Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104.
D000118:Acetylglucosaminidase / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002414:Cations, Monovalent; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D006596:Hexosaminidases / Q000378:metabolism*; D006867:Hydrolases / Q000378:metabolism*; D007700:Kinetics; D008247:Lysosomes / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000378:metabolism*; D012964:Sodium / Q000494:pharmacology; D013724:Teratoma
D002414:Cations, Monovalent; D012964:Sodium; D000242:Cyclic AMP; D006867:Hydrolases; D006596:Hexosaminidases; D000118:Acetylglucosaminidase; D010447:Peptide Hydrolases; D002118:Calcium