Comparison of the Changes in Protein Kinase C Induced by Glutamate in Primary Cortical Neurons and by in Vivo Cerebral Ischaemia

• Published in: Cellular signalling 1998-04, Vol.10 (4), p.291-295
• Main Authors: Chakravarthy, Balu R, Wang, Jian, Tremblay, Roger, Atkinson, Trevor G, Wang, Fuhu, Li, Hui, Buchan, Alastair M, Durkin, Jon P
• Format: Article
• Language: English
• Subjects: Animals; Brain Ischemia - enzymology; Brain Ischemia - physiopathology; Cells, Cultured; Cerebral Cortex - cytology; Cerebral Cortex - drug effects; Cerebral Cortex - enzymology; Enzyme Induction - drug effects; Excitatory Amino Acid Agonists - pharmacology; Glutamate toxicity; Glutamic Acid - toxicity; Hippocampal CA-1; Ischaemia; Male; Neurons - drug effects; Neurons - enzymology; Primary cortical cultures; Protein kinase C; Protein Kinase C - biosynthesis; Protein Kinase C - drug effects; Protein Kinase C - metabolism; Rats; Rats, Wistar
• ISSN: 0898-6568; 1873-3913
• DOI: 10.1016/S0898-6568(97)00131-9

Changes in protein kinase C (PKC) were compared in primary cortical neurons exposed to glutamate and in the CA-1 hippocampal region of rats subjected to transient cerebral ischaemia. After a 15-min exposure of cortical neurons to excitotoxic levels of glutamate, a 50–60% loss of membrane PKC activity but only about a 20 % loss in the amount of enzyme was observed, suggesting that in addition to enzyme loss other mechanisms also contributed to the overall loss of membrane PKC activity. Glutamate induced a 25–40% decrease in immunodetectable levels of PKC α, β, γ, and λ but no detectable changes in PCK ϵ and ζ. The loss of PKC activity coincided with a shift in electrophoretic mobility of PKC γ, ϵ, and λ, but not of PKC α, β, or ζ, suggesting post-translational modification of some PKC isoforms. By comparison, in rats subjected to transient (15-min) global ischaemia, a similar 50–60% decrease in membrane PKC activity, a 20–25 % loss in the amount of PKC, and a shift in PKC mobility were observed in CA-1 neurons 6 h post-reperfusion. In both the in vivo and the in vitro “ischaemic” models, administration of the AMPA receptor antagonist NBQX prevented the loss of PKC activity. These results indicate that the loss of PKC observed in in vivo ischaemia is likely to be due to excitotoxic damage and that this event can be closely mirrored in primary neuronal cultures damaged by glutamate.