Neuronal expression and regulation of CGRP promoter activity following viral gene transfer into cultured trigeminal ganglia neurons.
We have examined the regulation of calcitonin gene-related peptide (CGRP) promoter activity in primary cultures of rat trigeminal ganglia neurons. A viral vector was used to circumvent the potential complication of examining only a small subpopulation of cells in the heterogeneous cultures. Infection with high titers of recombinant adenovirus containing 1.25 kb of the rat CGRP promoter linked to the beta-galactosidase reporter gene (AdCGRP-lacZ) yielded expression in about 50% of the CGRP-expressing neurons. The CGRP-lacZ reporter gene was preferentially expressed in neurons, with 91% co-expression with endogenous CGRP. In contrast, an adenoviral vector containing a CMV-lacZ reporter was predominantly expressed in non-neuronal cells, with only 29% co-expression with CGRP. We then asked whether the CGRP promoter in the viral vector could be regulated by serotonin receptor type 1 (5-HT(1)) agonists. Promoter activity was decreased two- to threefold by treatment with five 5-HT(1B/D) agonists, including the triptan drugs sumatriptan, eletriptan, and rizatriptan that are used for migraine treatment. As controls, CMV promoter activity was not affected, and 5-HT(1B/D) receptor antagonists blocked the repression caused by sumatriptan and eletriptan. Thus, adenoviral gene transfer can be used in trigeminal ganglia neurons for studying the mechanisms of triptan drug action on CGRP synthesis.
Durham
Paul L
PL
Department of Biology, Southwest Missouri State University, Springfield, MO 65804, USA.
Dong
Penny X
PX
Belasco
Kevin T
KT
Kasperski
Jeffrey
J
Gierasch
William W
WW
Edvinsson
Lars
L
Heistad
Donald D
DD
Faraci
Frank M
FM
Russo
Andrew F
AF
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics*; D002452:Cell Count; D002478:Cells, Cultured; D004347:Drug Interactions; D005786:Gene Expression Regulation / Q000187:drug effects*; D017930:Genes, Reporter; D005814:Genes, Viral; D007150:Immunohistochemistry / Q000379:methods; D007211:Indoles / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology*; D012668:Trigeminal Ganglion / Q000166:cytology*; D014404:Tubulin / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D007211:Indoles; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; D014404:Tubulin; C007293:DAPI; D001616:beta-Galactosidase; D015740:Calcitonin Gene-Related Peptide