Gonzales_1987_J.Pharmacol.Exp.Ther_242_764

Alpha-1 receptor density and alpha-1 receptor-stimulated phosphoinositide hydrolysis in neuronal primary cultures are regulated by exposure of the cells to the alpha-1 agonist norepinephrine (NE). Pretreatment of neuronal cultures with 10 microM NE caused a time- and concentration-dependent decrease in NE-stimulated accumulation of [3H]inositol phosphates. The maximal NE-stimulated inositide hydrolysis was decreased by approximately 80% after 2 hr of pretreatment with NE, and this loss of responsiveness was reversible over a period of 12 hr. NE pretreatment of neuronal cultures did not affect the muscarinic receptor stimulation of inositide hydrolysis. Neither the number of alpha-1 receptor recognition sites, assessed by measuring the specific binding of DL-[125I]-alpha-(3-iodohydroxyphenyl)-ethyl-aminomethyl tetralone ([125I]HEAT), nor the Ki for NE inhibition of [125I]HEAT binding were changed appreciably after treating neurons with NE for 2 hr. A time- and concentration-dependent decrease in alpha-1 receptor recognition sites occurred with longer periods of NE pretreatment. The maximal loss of 50 to 60% [125I]HEAT binding sites was seen after 14 hr of pretreatment with NE. The number of [125I]HEAT binding sites returned to control levels within 24 hr in parallel with the recovery of alpha-1-stimulated inositide hydrolysis after being down-regulated by a 24-hr exposure to NE. Cycloheximide (3.5 microM) blocked both the recovery of alpha-1 receptors and the recovery of alpha-1-stimulated inositide hydrolysis after 24 hr of NE pretreatment.(ABSTRACT TRUNCATED AT 250 WORDS)