Role of second messengers in agonist up-regulation of 5-HT2A (5-HT2) receptor binding sites in cerebellar granule neurons: involvement of calcium influx and a calmodulin-dependent pathway.
The present study was undertaken to examine the involvement of 5-HT2A receptor-mediated second messengers in the agonist-induced up-regulation of 5-HT2A receptors in cerebellar granule cells. Stimulation of these cells with a 5-HT2A/5-HT2C agonist, (+/-)-(2, 5-dimethoxy-4-iodophenyl)-2-aminopropane for 16 hr resulted in a marked increase in [3H]ketanserin binding to 5-HT2A receptors in intact cells. (+/-)-(2, 5-dimethoxy-4-iodophenyl)-2- aminopropane up-regulated, but not basal levels of 5-HT2A binding sites, were largely attenuated by actinomycin D and cycloheximide, suggesting an essential role for de novo RNA and protein synthesis in this up-regulation process. This receptor up-regulation was 5-HT2A receptor-mediated but was unaffected by short-term pretreatment with phorbol dibutyrate to attenuate (+/-)-(2, 5-dimethoxy-4-iodophenyl)-2-aminopropane-induced phosphoinositide hydrolysis or by treatment with staurosporine to inhibit protein kinase C. In contrast, blockade of Ca2+ channels by LaCl3 or SK&F 96365 and depletion of extracellular Ca2+ by EGTA preferentially decreased the up-regulated 5-HT2A receptor levels, suggesting a role of Ca2+ influx in the receptor up-regulation. The (+/-)-(2, 5-dimethoxy-4- iodophenyl)-2-aminopropane up-regulation was also prevented by inhibitors of calmodulin, calmidazolium and W-7. Moreover, the effect of (+/-)-(2, 5-dimethoxy-4-iodophenyl)-2-aminopropane was blocked by KN-62, a selective Ca2+/calmodulin kinase inhibitor and by H-7 and staurosporine at concentrations high enough to be nonselective for a specific type of protein kinase.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D002118:Calcium / Q000502:physiology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000502:physiology; D002147:Calmodulin / Q000502:physiology*; D002478:Cells, Cultured; D002531:Cerebellum / Q000378:metabolism*; D010741:Phospholipases A / Q000502:physiology; D011493:Protein Kinase C / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011985:Receptors, Serotonin / Q000378:metabolism*; D015290:Second Messenger Systems / Q000502:physiology*; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000502:physiology; D015854:Up-Regulation
D002147:Calmodulin; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D010741:Phospholipases A; D010738:Type C Phospholipases; D002118:Calcium