Cyclic AMP increases the survival of ganglion cells in mixed retinal cell cultures in the absence of exogenous neurotrophic molecules, an effect that involves cholinergic activity.
Natural cell death is a well-known degenerative phenomenon occurring during development of the nervous system. The role of trophic molecules produced by target and afferent cells as well as by glial cells has been extensively demonstrated. Literature data demonstrate that cAMP can modulate the survival of neuronal cells. Cultures of mixed retinal cells were treated with forskolin (an activator of the enzyme adenylyl cyclase) for 48 h. The results show that 50 microM forskolin induced a two-fold increase in the survival of retinal ganglion cells (RGCs) in the absence of exogenous trophic factors. This effect was dose dependent and abolished by 1 microM H89 (an inhibitor of protein kinase A), 1.25 microM chelerythrine chloride (an inhibitor of protein kinase C), 50 microM PD 98059 (an inhibitor of MEK), 25 microM Ly 294002 (an inhibitor of phosphatidylinositol-3 kinase), 30 nM brefeldin A (an inhibitor of polypeptide release), and 10 microM genistein or 1 ng/ml herbimycin (inhibitors of tyrosine kinase enzymes). The inhibition of muscarinic receptors by 10 microM atropine or 1 microM telenzepine also blocked the effect of forskolin. When we used 25 microM BAPTA, an intracellular calcium chelator, as well as 20 microM 5-fluoro-2'-deoxyuridine, an inhibitor of cell proliferation, we also abolished the effect. Our results indicate that cAMP plays an important role controlling the survival of RGCs. This effect is directly dependent on M1 receptor activation indicating that cholinergic activity mediates the increase in RGC survival. We propose a model which involves cholinergic amacrine cells and glial cells in the increase of RGC survival elicited by forskolin treatment.
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Programa de Neuroimunologia, Departamento de Neurobiologia, Instituto de Biologia, Universidade Federal Fluminense, Caixo Postal 100.180, 24001-970 Niterói, RJ, Brazil.
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D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D005576:Colforsin / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D000242:Cyclic AMP / Q000494:pharmacology* / Q000502:physiology; D018727:Muscarinic Antagonists / Q000494:pharmacology*; D009457:Neuroglia / Q000187:drug effects* / Q000502:physiology; D018377:Neurotransmitter Agents / Q000494:pharmacology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D012160:Retina / Q000166:cytology; D012165:Retinal Ganglion Cells / Q000187:drug effects* / Q000502:physiology
D018727:Muscarinic Antagonists; D018377:Neurotransmitter Agents; D011500:Protein Synthesis Inhibitors; D005576:Colforsin; D000242:Cyclic AMP; D017853:Phosphotransferases (Alcohol Group Acceptor)