Ganglioside inhibition of glutamate-mediated protein kinase C translocation in primary cultures of cerebellar neurons.
In primary cultures of cerebellar granule cells, protein kinase C (PKC) translocation and activation can be triggered by the stimulation of excitatory amino acid neurotransmitter receptors. Glutamate evokes a dose-related translocation of 4-beta-[3H]phorbol 12,13-dibutyrate ([3H]-P(BtO)2) binding sites from the cytosol to the neuronal membrane and stimulates the incorporation of 32P into a number of membrane proteins, particularly protein bands in the range of 80, 50, and 40 kDa. The glutamate-evoked PKC translocation is Mg2+ sensitive, is prevented by 2-amino-5-phosphonovalerate and phencyclidine, is not inhibited by nitrendipine (a voltage-dependent Ca2+-channel blocker) but is abolished by the removal of Ca2+ from the incubation medium, suggesting that glutamate-mediated Ca2+ influx is operative in the redistribution of PKC. Exposure of granule cells to the gangliosides trisialosylgangliotetraglycosylceramide (GT1b) or monosialosylgangliotetraglycosylceramide (GM1) inhibits the translocation and activation of PKC evoked by glutamate. These glycosphingolipids fail to interfere with glutamate binding to its high-affinity recognition site or with the [3H]P(BtO)2 binding, nor do they affect the Ca2+ influx. These gangliosides may prevent PKC translocation by interfering with the PKC binding to the neuronal membrane phosphatidylserine.
Vaccarino
F
F
Fidia Georgetown Institute for the Neurosciences, Georgetown University Medical School, Washington, DC 20007.
Guidotti
A
A
Costa
E
E
D000818:Animals; D002118:Calcium / Q000502:physiology; D002451:Cell Compartmentation; D002478:Cells, Cultured; D002531:Cerebellum / Q000502:physiology*; D004789:Enzyme Activation; D018691:Excitatory Amino Acid Antagonists; D005732:Gangliosides / Q000494:pharmacology*; D005971:Glutamates / Q000502:physiology*; D008565:Membrane Proteins / Q000378:metabolism; D009568:Nitrendipine / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism; D011493:Protein Kinase C / Q000502:physiology*; D051381:Rats; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter / Q000502:physiology*; D013110:Sphingosine / Q000494:pharmacology*
D018691:Excitatory Amino Acid Antagonists; D005732:Gangliosides; D005971:Glutamates; D008565:Membrane Proteins; D010750:Phosphoproteins; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter; D009568:Nitrendipine; D011493:Protein Kinase C; D013110:Sphingosine; D002118:Calcium