Cyclic AMP regulates the expression of neurokinin1 receptors by neonatal rat spinal neurons in culture.
Neurokinin1 (NK1) receptors are up-regulated in the spinal cord during peripheral inflammation, but the biochemical mediators regulating this change have not been resolved. The promoter region of the gene encoding the NK1 receptor contains a cyclic AMP (cAMP)-responsive element. Therefore, we used primary cultures of neonatal rat spinal cord to test whether increasing intracellular cAMP can increase expression of NK1 receptors. Treatment with dibutyryl-cAMP (dbcAMP) resulted in a time-dependent increase in 125I-Bolton-Hunter-substance P (BHSP) binding in the cultures; treatment with dibutyryl-cyclic GMP did not. Treatment with forskolin plus 3-isobutyl-1-methylxanthine mimicked the increase in binding, providing further evidence for the involvement of cAMP in this effect. Scatchard analyses indicated that the increase in BHSP binding was due to an increase in binding capacity. The cAMP-induced increase in BHSP binding was preceded by an increase in levels of mRNA for NK1 receptor and was attenuated by pretreatment with cycloheximide. These data indicate that the cAMP-induced increase in binding was due to increased synthesis of NK1 receptors. Comparison of substance P (SP)-induced production of inositol phosphates between cultures pretreated with dbcAMP and controls suggested that increased expression of NK1 receptors did not result in increased generation of second messenger by NK1 receptor activation. Together, these data indicate that a persistent increase in intracellular cAMP increases expression of NK1 receptors. Because NK1 receptor activation contributes to increased excitability of spinal neurons, the increased expression of NK1 receptors may be important in maintaining responsiveness of spinal neurons to SP in central mechanisms underlying hyperalgesia.
Abrahams
L G
LG
Department of Cell Biology and Neuroanatomy, University of Minnesota, Minneapolis 55455, USA.
Reutter
M A
MA
McCarson
K E
KE
Seybold
V S
VS
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology*; D003513:Cycloheximide / Q000494:pharmacology; D003995:Dibutyryl Cyclic GMP / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D007295:Inositol Phosphates / Q000378:metabolism; D007457:Iodine Radioisotopes; D009474:Neurons / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D018040:Receptors, Neurokinin-1 / Q000096:biosynthesis / Q000235:genetics*; D013116:Spinal Cord / Q000378:metabolism*; D013373:Substance P / Q000378:metabolism; D013388:Succinimides / Q000378:metabolism
C043194:Bolton Hunter reagent-substance P conjugate; D007295:Inositol Phosphates; D007457:Iodine Radioisotopes; D012333:RNA, Messenger; D018040:Receptors, Neurokinin-1; D013388:Succinimides; D005576:Colforsin; D003995:Dibutyryl Cyclic GMP; D013373:Substance P; D003994:Bucladesine; D003513:Cycloheximide; D000242:Cyclic AMP; D015056:1-Methyl-3-isobutylxanthine