Regulation of acetylcholinesterase secretion from perfused bovine adrenal gland and isolated bovine chromaffin cells.
The secretion of acetylcholinesterase (AChE) was studied in an isolated perfused bovine adrenal gland preparation and in cultured bovine adrenal medullary chromaffin cells. Electrical field stimulation (10 Hz) of splanchnic nerve terminals in the isolated perfused gland resulted in a two-fold increase in AChE secretion from the gland. Perfusion with the cholinergic receptor antagonists mecamylamine (5 microM) and atropine (1 microM) inhibited 70% of the stimulated secretion of AChE, demonstrating that most of the stimulated secretion was derived from chromaffin cells. The effect of nicotine stimulation on the secretion of AChE from isolated bovine chromaffin cells was compared with that produced by other compounds (histamine, angiotensin II) which are known to stimulate secretion of catecholamines. Incubation with nicotine (1-25 microM) stimulated the secretion of catecholamines and AChE. Histamine (1 nM-10 microM) and angiotensin II (10 pM-10 microM) did not stimulate AChE secretion. Time-course studies of AChE resynthesis after irreversible inhibition with the esterase inhibitor diisopropylfluorophosphate (DFP) demonstrated that AChE is stored within chromaffin cells for at least 11 h before being secreted. AChE secretion was inhibited within 2-3 h by 10 micrograms/ml brefeldin A (BFA), a compound known to block protein translocation from the endoplasmic reticulum (ER) to the Golgi apparatus (GA). The results suggest that AChE may reside for 8-9 h within the lumen of the ER before being actively secreted by processing through the GA.
Small
D H
DH
Department of Pathology, University of Melbourne, Parkville, Victoria, Australia.
Michaelson
S
S
Marley
P D
PD
Friedhuber
A
A
Hocking
A
A
Livett
B G
BG
D000110:Acetylcholinesterase / Q000378:metabolism*; D000311:Adrenal Glands / Q000378:metabolism*; D000818:Animals; D020126:Brefeldin A; D002417:Cattle; D002469:Cell Separation; D002838:Chromaffin System / Q000166:cytology / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology; D004558:Electric Stimulation; D007531:Isoflurophate / Q000494:pharmacology; D008854:Microscopy, Electron; D010477:Perfusion
D003517:Cyclopentanes; D007531:Isoflurophate; D020126:Brefeldin A; D000110:Acetylcholinesterase