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Down regulation of protein kinase C in neuronal cells: effects on neurotransmitter release

HJ Matthies, HC Palfrey, LD Hirning and RJ Miller
Journal of Neuroscience 1 April 1987, 7 (4) 1198-1206; https://doi.org/10.1523/JNEUROSCI.07-04-01198.1987
HJ Matthies
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HC Palfrey
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LD Hirning
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RJ Miller
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Abstract

We investigated the effects of phorbol esters on protein kinase C (PKC) activity and on neurotransmitter release from cultured neuronal cells. Both differentiated and undifferentiated PC12 pheochromocytoma cells contained high levels of protein PKC. Under normal conditions all the enzyme activity was found in the cytoplasm. Addition of the phorbol esters phorbol 12-myristate-13-acetate (TPA) or phorbol 12,13- dibutyrate (PDBu) caused a rapid translocation of PKC from the cytoplasm to the particulate fraction. Continued culture of cells with these phorbol esters resulted in the decline of total PKC activity. After 10–20 hr of culture, both membrane and cytoplasmic PKC activity had declined to background levels. cAMP-dependent and Ca2+/calmodulin- dependent protein kinase activities were only slightly affected by chronic phorbol ester treatment. Addition of active phorbol esters to PC12 cells produced an enhancement of the depolarization-induced release of 3H-norepinephrine. Following chronic phorbol ester treatment, the ability of these substances to enhance evoked catecholamine release was lost. Furthermore, depolarizing stimuli released considerably less 3H-norepinephrine than in control untreated cells. Phorbol esters also enhanced depolarization-induced 3H- norepinephrine release from primary cultures of rat sympathetic neurons. Chronic treatment of these neurons with phorbol esters also resulted in the loss of their ability to enhance transmitter release and in a large reduction in the extent of depolarization-evoked transmitter release. Chronic phorbol ester treatment also resulted in the disappearance of PKC from sympathetic neurons, but had little effect on cAMP-dependent or Ca2+/calmodulin-dependent kinase activities. These results demonstrate that PKC-deficient neurons can be prepared. The data also demonstrate that depolarization-induced neurotransmitter release is mediated by both protein kinase C-dependent and independent pathways.

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The Journal of Neuroscience: 7 (4)
Journal of Neuroscience
Vol. 7, Issue 4
1 Apr 1987
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Down regulation of protein kinase C in neuronal cells: effects on neurotransmitter release
HJ Matthies, HC Palfrey, LD Hirning, RJ Miller
Journal of Neuroscience 1 April 1987, 7 (4) 1198-1206; DOI: 10.1523/JNEUROSCI.07-04-01198.1987

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Down regulation of protein kinase C in neuronal cells: effects on neurotransmitter release
HJ Matthies, HC Palfrey, LD Hirning, RJ Miller
Journal of Neuroscience 1 April 1987, 7 (4) 1198-1206; DOI: 10.1523/JNEUROSCI.07-04-01198.1987
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