Vesicle-mediated phosphatidylcholine reapposition to the plasma membrane following hormone-induced phospholipase D activation.
Phospholipase D (PLD) activation involved in signal transduction may lead to the hydrolysis of conspicuous amounts of phosphatidylcholine (PC). This study shows that PLD activation significantly alters the plasma membrane (PM) environment and the membrane exchange dynamics. PC-PLD activation in vasopressin (AVP)-stimulated L6 myogenic cells was accompanied by increased exocytosis and decreased membrane fluidity, as shown by transmission EM and fluorescence spectroscopy of trimethylammonium-diphenyl-hexatriene. AVP-induced exocytosis appeared to be brefeldin A-insensitive. PLD inhibition by Zn(2+) and PC de novo synthesis inhibition by hexadecylphosphocholine abolished AVP-induced vesicle traffic. Upon AVP stimulation, metabolically labeled PC decreased in PM, then transiently increased in microsomes, and returned to the prestimulus level in the PM within 5 min, a phenomenon requiring PC neosynthesis and microtubule functionality. Vesicle traffic with similar features was also observed after endothelin-1-induced PC-PLD activation in rat peritubular myoid cells. These results indicate that, in nonsecretory cells, exocytosis coupled to PC de novo synthesis restores PM-PC, conspicuously consumed during PLD-mediated signal transduction.
Coletti
D
D
Department of Histology and Medical Embryology, University "La Sapienza,", Rome, Italy.
Silvestroni
L
L
Naro
F
F
Molinaro
M
M
Adamo
S
S
Palleschi
S
S
D020106:Acrylamide / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001127:Arginine Vasopressin / Q000494:pharmacology*; D020126:Brefeldin A / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D003571:Cytochalasin B / Q000494:pharmacology; D004161:Diphenylhexatriene / Q000031:analogs & derivatives; D004789:Enzyme Activation; D005089:Exocytosis; D005456:Fluorescent Dyes; D007700:Kinetics; D008563:Membrane Lipids / Q000378:metabolism; D018482:Muscle, Skeletal; D015739:Nocodazole / Q000494:pharmacology; D017239:Paclitaxel / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000378:metabolism*; D010739:Phospholipase D / Q000378:metabolism*; D051381:Rats; D000077191:Wortmannin; D015032:Zinc / Q000494:pharmacology
D000730:Androstadienes; D005456:Fluorescent Dyes; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D001127:Arginine Vasopressin; D004161:Diphenylhexatriene; D020126:Brefeldin A; D020106:Acrylamide; D003571:Cytochalasin B; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; D010739:Phospholipase D; D015032:Zinc; D017239:Paclitaxel; D015739:Nocodazole; D000077191:Wortmannin