The involvement of protein kinase C in the regulation of choline cotransport in Limulus.
The involvement of protein kinase C (PKC) in the regulation of [3H]choline cotransport was studied in Limulus brain hemi-slice preparations. The PKC activators, phorbol 12-myristate 13-acetate (PMA) or phorbol 12,13-dibutyrate (PDBu), significantly decreased [3H]choline cotransport. Conversely, the PKC inhibitors, staurosporine (STAURO) and polymyxin B (PMB), each increased [3H]choline cotransport. These PKC inhibitors prevented the phorbol ester-induced reduction of transport. Both the PMA induced decrease and the STAURO induced increase in [3H]choline cotransport were paralleled by respective and comparable changes in [3H]hemicholinium-3 (HC-3) specific binding. Pre-exposure of brain hemi-slices to elevated potassium chloride (120 mM KCl) resulted in a doubling of [3H]choline cotransport and [3H]HC-3 binding. The enhancement of [3H]choline cotransport by STAURO and antecedent 120 mM KCl treatment were additive. PMA did not significantly alter elevated potassium stimulated transport. Moreover, arachidonyltrifluoromethyl ketone (AACOCF3) and quinacrine (QUIN), both phospholipase A2 (PLA2) inhibitors, markedly decreased enhanced [3H]choline transport and [3H]HC-3 binding induced by antecedent exposure to depolarizing concentrations of potassium. These results suggest that PKC and PLA2 are involved in the regulation of [3H]choline cotransport but at different regulatory sites.
Ford
B D
BD
Department of Anatomy and Physiology, Meharry Medical College, Nashville, TN 37208, USA.
Ivy
M T
MT
Mtshali
C P
CP
Townsel
J G
JG
D000818:Animals; D001095:Arachidonic Acids / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D001921:Brain / Q000201:enzymology; D002273:Carcinogens / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D002794:Choline / Q000493:pharmacokinetics*; D018678:Cholinergic Agents / Q000493:pharmacokinetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006426:Hemicholinium 3 / Q000493:pharmacokinetics; D006737:Horseshoe Crabs / Q000201:enzymology*; D009420:Nervous System / Q000201:enzymology; D009924:Organ Culture Techniques; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D011189:Potassium Chloride / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D011796:Quinacrine / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014316:Tritium
D001095:Arachidonic Acids; D002273:Carcinogens; D002352:Carrier Proteins; D018678:Cholinergic Agents; D004791:Enzyme Inhibitors; C081565:arachidonyltrifluoromethane; D014316:Tritium; D006426:Hemicholinium 3; D015240:Phorbol 12,13-Dibutyrate; D011189:Potassium Chloride; D011493:Protein Kinase C; D010741:Phospholipases A; D054467:Phospholipases A2; D011796:Quinacrine; D019311:Staurosporine; D002794:Choline; D013755:Tetradecanoylphorbol Acetate