Preferential release of catecholamine from permeabilized PC12 cells by alpha- and beta-type protein kinase C subspecies.
Protein kinase C (PKC) is now recognized as comprising two groups of closely related subspecies. The PKC gamma enzyme is apparently present only in central nervous tissues, and hence was expected to participate in neurotransmitter release. We have utilized a 'depletion-insertion' method to identify the PKCs participating in the exocytotic response. PC12 cells were 'down-regulated' by prior treatment (24 h) with phorbol 12-myristate 13-acetate (PMA; 1 microM), which nearly abolished endogenous PKC activity. Down-regulated PC12 cells were loaded with [3H]dopamine, permeabilized with digitonin, and recombinant or purified PKCs were inserted and activated with a low dose of PMA (20 nM). Among group A PKCs, PKC alpha was the most effective activator of [3H]dopamine release (215%), followed by beta II (185%) and beta I (150%). PKC gamma had no consistent effect on neurotransmitter release. PC12 cells express PKC alpha and PKC beta, but not PKC gamma, as revealed by Northern-blot analysis. We therefore postulate that PKC alpha and PKC beta participate in neurotransmitter release, whereas PKC gamma might be involved in other neuronal functions.
Ben-Shlomo
H
H
Department of Biochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel.
Sigmund
O
O
Stabel
S
S
Reiss
N
N
Naor
Z
Z
D000818:Animals; D015152:Blotting, Northern; D002463:Cell Membrane Permeability; D015342:DNA Probes; D004072:Digitonin / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D005089:Exocytosis; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D016716:PC12 Cells; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D014316:Tritium
D015342:DNA Probes; D007527:Isoenzymes; D012334:RNA, Neoplasm; D014316:Tritium; D011493:Protein Kinase C; D004072:Digitonin; D004298:Dopamine