Functional characterization of rat submaxillary gland muscarinic receptors using microphysiometry.
1. Muscarinic cholinoceptors (MChR) in freshly dispersed rat salivary gland (RSG) cells were characterized using microphysiometry to measure changes in acidification rates. Several non-selective and selective muscarinic antagonists were used to elucidate the nature of the subtypes mediating the response to carbachol. 2. The effects of carbachol (pEC(50) = 5.74 +/- 0.02 s.e.mean; n = 53) were highly reproducible and most antagonists acted in a surmountable, reversible fashion. The following antagonist rank order, with apparent affinity constants in parentheses, was noted: 4-DAMP (8.9)= atropine (8.9) > tolterodine (8.5) > oxybutynin (7.9) > S-secoverine (7.2) > pirenzepine (6.9) > himbacine (6.8) > AQ-RA 741 (6.6) > methoctramine (5.9). 3. These studies validate the use of primary isolated RSG cells in microphysiometry for pharmacological analysis. These data are consistent with, and extend, previous studies using alternative functional methods, which reported a lack of differential receptor pharmacology between bladder and salivary gland tissue. 4. The antagonist affinity profile significantly correlated with the profile at human recombinant muscarinic M(3) and M(5) receptors. Given a lack of antagonists that discriminate between M(3) and M(5), definitive conclusion of which subtype(s) is present within RSG cells cannot be determined.
Neurobiology Unit, Roche Bioscience, 3401 Hillview Avenue, Palo Alto, California, CA 94304, USA. trena.meloy@roche.com
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Meloy
D V
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Daniels
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Hegde
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Eglen
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Ford
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D001559:Benzhydryl Compounds / Q000494:pharmacology; D001570:Benzodiazepinones / Q000494:pharmacology; D001667:Binding, Competitive / Q000187:drug effects; D015374:Biosensing Techniques; D002217:Carbachol / Q000378:metabolism / Q000494:pharmacology; D018679:Cholinergic Agonists / Q000378:metabolism / Q000494:pharmacology; D003408:Cresols / Q000494:pharmacology; D003959:Diamines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005663:Furans; D008297:Male; D008333:Mandelic Acids / Q000494:pharmacology; D018727:Muscarinic Antagonists / Q000494:pharmacology; D009281:Naphthalenes; D010627:Phenethylamines / Q000494:pharmacology; D010665:Phenylpropanolamine; D010880:Piperidines / Q000494:pharmacology; D010890:Pirenzepine / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism*; D013363:Submandibular Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000068737:Tolterodine Tartrate
D000470:Alkaloids; D001559:Benzhydryl Compounds; D001570:Benzodiazepinones; D018679:Cholinergic Agonists; D003408:Cresols; D003959:Diamines; D005663:Furans; D008333:Mandelic Acids; D018727:Muscarinic Antagonists; D009281:Naphthalenes; D010627:Phenethylamines; D010880:Piperidines; D011976:Receptors, Muscarinic; C069054:AQ-RA 741; D010665:Phenylpropanolamine; D010890:Pirenzepine; C028187:secoverine; D000068737:Tolterodine Tartrate; D001285:Atropine; C042375:4-diphenylacetoxy-1,1-dimethylpiperidinium; D002217:Carbachol; C005419:oxybutynin; C048172:himbacine; C054938:methoctramine