title: Proliferation of guinea pig tracheal epithelial cells in coculture with rat dorsal root ganglion neural cells.
abstract: Neuropeptides secreted by sensory afferent nerves in airways may modulate growth of airway epithelial cells. To determine whether airway sensory C-fiber nerves secrete neuropeptides that stimulate airway epithelial cell proliferation, we measured S-phase traversal in guinea pig tracheal epithelial (GPTE) cells after coculture with rat dorsal root ganglion (DRG) cells. GPTE cells were grown in subconfluent culture on collagen-coated filters for 2 days. DRG cells were harvested from newborn rat pups and grown in primary culture for 7-10 days in separate wells. GPTE and DRG cells then were cocultured for 48 h, and 10 mM bromodeoxyuridine (BrdU), a thymidine analogue, was added in the final 24 h. Control GPTE cells were grown under similar conditions but without DRG cells. Coculture with DRG cells stimulated GPTE cell traversal of S phase. BrdU labeling in cocultured GPTE cells was 42.8 +/- 5.8 compared with 18.1 +/- 7.2% in control GPTE cells (P < 0.001, n = 6). Coculture in the presence of either the neurokinin (NK)1 receptor antagonists LY-297911 or CP-99,994, the NK2 receptor antagonist SR-48,968, or the calcitonin gene-related peptide (CGRP) receptor antagonist hCGRP-(8-37) (10(-7) M of each) during coculture attenuated proliferation of GPTE cells. Treatment with all three antagonists together during coculture decreased BrdU labeling to 2.4 +/- 0.9% of labeled cells vs. 8.5 +/- 0.5% of labeled cells during coculture without antagonists (n = 4, P < 0.02). DRG cells in coculture secreted substantial concentrations of CGRP [71.0 +/- 11.3 (+/- SE) pmol/ml], substance P (1.26 +/- 0.35 pmol/ml), and neurokinin A (0.45 +/- 0.10 pmol/ml) (n = 19 for each).(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Medicine, University of Chicago, Illinois 60637, USA.
authors.forename: S R
authors.initials: SR
authors.lastname: White
authors.forename: A
authors.initials: A
authors.lastname: Garland
authors.forename: B
authors.initials: B
authors.lastname: Gitter
authors.forename: I
authors.initials: I
authors.lastname: Rodger
authors.forename: L E
authors.initials: LE
authors.lastname: Alger
authors.forename: J
authors.initials: J
authors.lastname: Necheles
authors.forename: A R
authors.initials: AR
authors.lastname: Nawrocki
authors.forename: J
authors.initials: J
authors.lastname: Solway
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001549:Benzamides / Q000494:pharmacology; D001973:Bromodeoxyuridine; D015740:Calcitonin Gene-Related Peptide / Q000494:pharmacology; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D002450:Cell Communication; D002453:Cell Cycle; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology*; D006168:Guinea Pigs; D015288:Neurokinin A / Q000037:antagonists & inhibitors; D064729:Neurokinin-1 Receptor Antagonists; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D010446:Peptide Fragments / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018039:Receptors, Tachykinin / Q000037:antagonists & inhibitors; D016196:S Phase; D014132:Trachea / Q000166:cytology* / Q000502:physiology
chemical_list: D001549:Benzamides; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D064729:Neurokinin-1 Receptor Antagonists; D010446:Peptide Fragments; D010880:Piperidines; D018039:Receptors, Tachykinin; C058636:calcitonin gene-related peptide (8-37); C080755:3-(2-methoxybenzylamino)-2-phenylpiperidine; C073839:SR 48968; D015288:Neurokinin A; D001973:Bromodeoxyuridine; D015740:Calcitonin Gene-Related Peptide