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Combined antisense and pharmacological approaches implicate hTASK as an airway O(2) sensing K(+) channel

Combined antisense and pharmacological approaches implicate hTASK as an airway O(2) sensing K(+) channel
Combined antisense and pharmacological approaches implicate hTASK as an airway O(2) sensing K(+) channel
Neuroepithelial bodies act as airway oxygen sensors. The lung carcinoma line H146 is an established model for neuroepithelial body cells. Although O(2) sensing in both cells is via NADPH oxidase H(2)O(2)/free radical production and acute hypoxia promotes K(+) channel closure and cell depolarization, the identity of the K(+) channel is still controversial. However, recent data point toward the involvement of a member of the tandem P domain family of K(+) channels. Reverse transcription-polymerase chain reaction screening indicates that all known channels other than hTWIK1 and hTRAAK are expressed in H146 cells. Our detailed pharmacological characterization of the O(2)-sensitive K(+) current described herein is compatible with the involvement of hTASK1 or hTASK3 (pH dependence, tetraethylammonium and dithiothreitol insensitivity, blockade by arachidonic acid, and halothane activation). Furthermore, we have used antisense oligodeoxynucleotides directed against hTASK1 and hTASK3 to suppress almost completely the hTASK1 protein and show that these cells no longer respond to acute hypoxia; this behavior was not mirrored in liposome-only or missense-treated cells. Finally, we have used Zn(2+) treatment as a maneuver able to discriminate between these two homologues of hTASK and show that the most likely candidate channel for O(2) sensing in these cells is hTASK3.
dithiothreitol, drug effects, tandem pore domain, carcinoma, research support, anoxia, metabolism, nerve tissue proteins, oligonucleotides, proteins, lung, family, respiratory system, potassium channels, halothane, nerve tissue, nadph oxidase, research, acid, treatment, oxygen, humans, protein, antisense, analysis, genetics, behavior, potassium, arachidonic acid
0021-9258
26499-26508
Hartness, M.E.
72efd180-4428-4ee6-b59c-a90bc3d2605e
Lewis, A.
99987827-edf2-4157-b60b-81f262505bd0
Searle, G.J.
75d1b72a-b229-42cd-8fc2-c834764c3276
O'Kelly, I.
e640f28a-42f0-48a6-9ce2-cb5a85d08c66
Peers, C.
25d5cc41-76e1-43b6-8759-223e06ba1f5f
Kemp, P.J.
954a798c-02bd-40e9-beb6-ad12df9613ed
Hartness, M.E.
72efd180-4428-4ee6-b59c-a90bc3d2605e
Lewis, A.
99987827-edf2-4157-b60b-81f262505bd0
Searle, G.J.
75d1b72a-b229-42cd-8fc2-c834764c3276
O'Kelly, I.
e640f28a-42f0-48a6-9ce2-cb5a85d08c66
Peers, C.
25d5cc41-76e1-43b6-8759-223e06ba1f5f
Kemp, P.J.
954a798c-02bd-40e9-beb6-ad12df9613ed

Hartness, M.E., Lewis, A., Searle, G.J., O'Kelly, I., Peers, C. and Kemp, P.J. (2001) Combined antisense and pharmacological approaches implicate hTASK as an airway O(2) sensing K(+) channel. The Journal of Biological Chemistry, 276 (28), 26499-26508. (doi:10.1074/jbc.M010357200).

Record type: Article

Abstract

Neuroepithelial bodies act as airway oxygen sensors. The lung carcinoma line H146 is an established model for neuroepithelial body cells. Although O(2) sensing in both cells is via NADPH oxidase H(2)O(2)/free radical production and acute hypoxia promotes K(+) channel closure and cell depolarization, the identity of the K(+) channel is still controversial. However, recent data point toward the involvement of a member of the tandem P domain family of K(+) channels. Reverse transcription-polymerase chain reaction screening indicates that all known channels other than hTWIK1 and hTRAAK are expressed in H146 cells. Our detailed pharmacological characterization of the O(2)-sensitive K(+) current described herein is compatible with the involvement of hTASK1 or hTASK3 (pH dependence, tetraethylammonium and dithiothreitol insensitivity, blockade by arachidonic acid, and halothane activation). Furthermore, we have used antisense oligodeoxynucleotides directed against hTASK1 and hTASK3 to suppress almost completely the hTASK1 protein and show that these cells no longer respond to acute hypoxia; this behavior was not mirrored in liposome-only or missense-treated cells. Finally, we have used Zn(2+) treatment as a maneuver able to discriminate between these two homologues of hTASK and show that the most likely candidate channel for O(2) sensing in these cells is hTASK3.

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More information

Published date: 2001
Keywords: dithiothreitol, drug effects, tandem pore domain, carcinoma, research support, anoxia, metabolism, nerve tissue proteins, oligonucleotides, proteins, lung, family, respiratory system, potassium channels, halothane, nerve tissue, nadph oxidase, research, acid, treatment, oxygen, humans, protein, antisense, analysis, genetics, behavior, potassium, arachidonic acid

Identifiers

Local EPrints ID: 59814
URI: http://eprints.soton.ac.uk/id/eprint/59814
ISSN: 0021-9258
PURE UUID: 800ba672-c28f-4c22-a870-0d16fcf77edb

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Date deposited: 04 Sep 2008
Last modified: 15 Mar 2024 11:17

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Contributors

Author: M.E. Hartness
Author: A. Lewis
Author: G.J. Searle
Author: I. O'Kelly
Author: C. Peers
Author: P.J. Kemp

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