Neuroprotective effects of modulators of P2 receptors in primary culture of CNS neurones.
In previous studies (Volonté and Merlo, 1996. J. Neurosci. Res. 45, 183-193) basilen blue was shown to be a P2 receptor antagonist which abrogated glutamate-mediated cytotoxicity in cerebellar neurones in primary culture. Our work has now been extended to evaluate the neuroprotective action of the compound in additional neuronal systems, as well as in a different paradigm of cell death. We show that basilen blue prevents L-glutamate-mediated neurotoxicity in rat cerebellar (90-100% inhibition), cortical (60-70%) and hippocampal (50%) neurones. Similarly, glutamate-dependent progressive darkening of cell bodies, loss of phase-brightness and rapid cellular swelling are inhibited. Basilen blue is significantly less toxic and more effective at blocking L-glutamate toxicity in mixed cortical/glial cultures, compared to its structural analogue cibacron blue. Moreover, its neuroprotective effect is correlated with the time of incubation with granule neurones. Other purinoceptor ligands, including 2,2'-pyridylisatogen, but not pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid 4-sodium, are also effective in preventing glutamate toxicity. Furthermore, basilen blue prevents serum deprivation- and low potassium-induced apoptotic cell death in cerebellar granule neurones. In summary, our data extend and reinforce the possibility of a potential therapeutic use of P2 receptor modulators as neuroprotective agents for the central nervous system.
Volonté
C
C
Institute of Neurobiology, C.N.R., I.R.C.C.S. Santa Lucia, Rome, Italy. cinzia@kant.irmkant.rm.cnr.it
Ciotti
M T
MT
D'Ambrosi
N
N
Lockhart
B
B
Spedding
M
M
D000818:Animals; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology; D002540:Cerebral Cortex / Q000166:cytology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology; D007510:Isatin / Q000031:analogs & derivatives / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D014227:Triazines / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D018696:Neuroprotective Agents; D018048:Receptors, Purinergic P2; D014227:Triazines; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; C016708:2,2'-pyridylisatogen; D018698:Glutamic Acid; C012842:Cibacron Blue F 3GA; D011732:Pyridoxal Phosphate; D007510:Isatin