Cytochemical demonstration of NPPase activity for detecting proton-translocating ATPase of Golgi complex in rat pancreatic acinar cells.
An attempt at cytochemical demonstration of acidification proton-translocating ATPase (H(+)-ATPase) of Golgi complex in rat pancreatic acinar cells has been made by using p-nitrophenylphosphatase (NPPase) cytochemistry which is used for detecting of Na(+)-K(+)-ATPase (Mayahara et al. 1980) and gastric H(+)-K(+)-ATPase (Fujimoto et al. 1986). K(+)-independent NPPase activity was observed on the membrane of the trans cisternae of Golgi complex, but not inside of cisternae. The localization of NPPase activity is different from that of acid phosphatase activity where reaction products were seen on the inside of the trans Golgi cisternae. Since this activity was insensitive to vanadate, ouabain and independent of potassium ions, it was distinct from plasma membranous ATPases such as Na(+)-K(+)-ATPase and Ca2(+)-ATPase. The K(+)-independent NPPase activity was diminished by the inhibitors of H(+)-ATPase such as N-ethylmaleimide (NEM) and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). The NPPase reaction products were also seen on the membranes of other acidic organelles, i.e., lysosomes, endosomes, autophagosomes and coated vesicles. These results suggest that NPPase activity on the membrane of the Golgi complex and other acidic organelles corresponds with H(+)-ATPase which plays a role in acidification.
Araki
N
N
Department of Anatomy, Ehime University School of Medicine, Japan.
Lee
M
M
Takashima
Y
Y
Ogawa
K
K
D000818:Animals; D002951:Citrates; D019343:Citric Acid; D006056:Golgi Apparatus / Q000201:enzymology*; D006651:Histocytochemistry; D009596:Nitrophenols / Q000378:metabolism*; D009943:Organophosphorus Compounds / Q000378:metabolism*; D010179:Pancreas / Q000201:enzymology* / Q000648:ultrastructure; D011188:Potassium / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000032:analysis* / Q000037:antagonists & inhibitors; D051381:Rats; D011919:Rats, Inbred Strains
D002951:Citrates; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D019343:Citric Acid; C008644:nitrophenylphosphate; D006180:Proton-Translocating ATPases; D011188:Potassium