Neurotoxicity of HIV coat protein gp120, NMDA receptors, and protein kinase C: a study with rat cerebellar granule cell cultures.
The neuronal loss observed in AIDS patients may be partly due to the neurotoxicity of HIV coat protein gp120, whose mechanism of action has been suggested to involve an interaction with voltage-dependent Ca2+ channels and NMDA receptors (Lipton, Trends Neurosci 15:75-79, 1992). In the present investigation we analyzed the acute neurotoxicity of gp120 on a purified neuronal population (rat cerebellar granule cell cultures) amply used for studies on glutamate toxicity. Cultures of 7-8 days were exposed for 15 min to a buffered Locke's solution containing the substances under study, washed, and cultured for another 24 hr in their original medium. The cells were stained with the nuclear dyes propidium iodide (for dead cells) and Hoechst 33258 (for total cells) and counted. Average cell death in controls was 8%. gp120 (1 pM-10 nM) caused an increase of cell death of about 80%. The effect was totally antagonized by NMDA antagonists (1 mM APV and 10 microM MK-801), by 1 microM nifedipine, and by anti-gp120 antibodies. At a concentration of 100 microM glutamate caused an average 130% increase of cell death, which was totally antagonized by APV. The effect of gp120 or glutamate did not appear to be mediated by the secretion of neurotoxins by nonneuronal cells present in a low proportion in the cultures nor to be due to the inactivation of (or competition with) neurotrophic factors present in the medium. The simultaneous administration of gp120 and glutamate (in various combinations of concentrations) had an effect that was less than additive.(ABSTRACT TRUNCATED AT 250 WORDS)
Savio
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T
Neurobiology Section, Istituto Superiore di Sanità, Roma, Italy.
Levi
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D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D003594:Cytoplasmic Granules / Q000502:physiology; D004357:Drug Synergism; D005971:Glutamates / Q000633:toxicity; D018698:Glutamic Acid; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000633:toxicity*; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D010703:Phorbol Esters / Q000378:metabolism / Q000633:toxicity; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects*; D013194:Staining and Labeling; D014178:Translocation, Genetic / Q000187:drug effects
D005971:Glutamates; D015699:HIV Envelope Protein gp120; D010703:Phorbol Esters; D016194:Receptors, N-Methyl-D-Aspartate; D015240:Phorbol 12,13-Dibutyrate; D018698:Glutamic Acid; D011493:Protein Kinase C