Characterization of the Kv channels of mouse carotid body chemoreceptor cells and their role in oxygen sensing.
As there are wide interspecies variations in the molecular nature of the O(2)-sensitive Kv channels in arterial chemoreceptors, we have characterized the expression of these channels and their hypoxic sensitivity in the mouse carotid body (CB). CB chemoreceptor cells were obtained from a transgenic mouse expressing green fluorescent protein (GFP) under the control of tyrosine hydroxylase (TH) promoter. Immunocytochemical identification of TH in CB cell cultures reveals a good match with GFP-positive cells. Furthermore, these cells show an increase in [Ca(2+)](i) in response to low P(O(2)), demonstrating their ability to engender a physiological response. Whole-cell experiments demonstrated slow-inactivating K(+) currents with activation threshold around -30 mV and a bi-exponential kinetic of deactivation (tau of 6.24 +/- 0.52 and 32.85 +/- 4.14 ms). TEA sensitivity of the currents identified also two different components (IC(50) of 17.8 +/- 2.8 and 940.0 +/- 14.7 microm). Current amplitude decreased reversibly in response to hypoxia, which selectively affected the fast deactivating component. Hypoxic inhibition was also abolished in the presence of low (10-50 microm) concentrations of TEA, suggesting that O(2) interacts with the component of the current most sensitive to TEA. The kinetic and pharmacological profile of the currents suggested the presence of Kv2 and Kv3 channels as their molecular correlates, and we have identified several members of these two subfamilies by single-cell PCR and immunocytochemistry. This report represents the first functional and molecular characterization of Kv channels in mouse CB chemoreceptor cells, and strongly suggests that O(2)-sensitive Kv channels in this preparation belong to the Kv3 subfamily.
Pérez-García
M Teresa
MT
Universidad de Valladolid y Consejo Superior de Investigaciones Cientificas (CSIC), Departamento de Bioquímica y Biología Molecular y Fisiología, Facultad de Medicina, Valladolid, Spain. tperez@ibgm.uva.es
Colinas
Olaia
O
Miguel-Velado
Eduardo
E
Moreno-Domínguez
Alejandro
A
López-López
José Ramón
JR
D000818:Animals; D002344:Carotid Body / Q000737:chemistry / Q000378:metabolism*; D015687:Cell Hypoxia; D002478:Cells, Cultured; D002628:Chemoreceptor Cells / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis; D051379:Mice; D008822:Mice, Transgenic; D018408:Patch-Clamp Techniques; D024642:Potassium Channels, Voltage-Gated / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051670:Shaw Potassium Channels; D019789:Tetraethylammonium / Q000494:pharmacology; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis / Q000096:biosynthesis
C490839:Kcnc1 protein, mouse; D024642:Potassium Channels, Voltage-Gated; D021122:Protein Subunits; D012333:RNA, Messenger; D051670:Shaw Potassium Channels; D049452:Green Fluorescent Proteins; D019789:Tetraethylammonium; D014446:Tyrosine 3-Monooxygenase