Adenylyl cyclase is involved in desensitization and recovery of ATP-stimulated Cl- secretion in MDCK cells.
We investigated the process of and recovery from desensitization of the P2 receptor-mediated stimulation of Cl- secretion in Madin-Darby canine kidney (MDCK) cell monolayers by assaying the response of short-circuit current (Isc). When the cells were exposed to repeated 3-min challenges of ATP or UTP interspersed with 5-min washes, the response of Isc desensitized rapidly followed by spontaneous recovery. The pattern of inhibition by various channel blockers or enzyme inhibitors revealed that both the initial and recovered responses of Isc have the same ionic and signaling mechanisms. The desensitization and recovery processes were confined to the membrane exposed to the repeated challenges. When added during the desensitized phase, 8-bromoadenosine 3',5'-cyclic monophosphate enhanced the ATP-stimulated Isc response, whereas it did not during the initial or recovered phases. ATP-induced increases of intracellular adenosine 3',5'-cyclic monophosphate showed similar desensitization and recovery in parallel with the changes in the responses of Isc. The desensitization process was attenuated by pretreatment with cholera toxin or pertussis toxin. Taken together, our results suggest that the adenylyl cyclase system plays a role in the desensitization and recovery mechanism of the ATP-stimulated Cl- secretion in MDCK cells.
Woo
J S
JS
Department of Physiology and Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Inoue
C N
CN
Hanaoka
K
K
Schwiebert
E M
EM
Guggino
S E
SE
Guggino
W B
WB
D000255:Adenosine Triphosphate / Q000502:physiology*; D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology*; D002712:Chlorides / Q000378:metabolism*; D002772:Cholera Toxin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology; D004285:Dogs; D015640:Ion Channel Gating / Q000502:physiology; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D037342:Pertussis Toxin; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D037361:Adenylate Cyclase Toxin; D018118:Chloride Channels; D002712:Chlorides; D010566:Virulence Factors, Bordetella; D000255:Adenosine Triphosphate; D002772:Cholera Toxin; D000242:Cyclic AMP; D037342:Pertussis Toxin; D000262:Adenylyl Cyclases