Probing molecular identity of native single potassium channels by overexpression of dominant negative subunits.
Overexpression of dominant negative subunits previously has been shown to affect the whole cell delayed-rectifier potassium current (Ikv) in Xenopus spinal neurons. Here, we show that effects of overexpression of wild-type and dominant negative Kv1 channels are evident at the single channel level. The goal of these studies was to match molecular species of Kv subunits to specific, functionally identified single voltage-dependent potassium channels. In a heterologous system (the Xenopus oocyte), co-expression of wild-type and dominant negative mutant Kv1.1 subunits results in loss of active channels rather than channels of altered conductance. However, in situ overexpression studies are difficult to interpret due to the diversity in the control population of channels. Therefore, identification of endogenous channel populations containing Kv1 subunits is limited. Future work will reduce the endogenous diversity of potassium channels by study of single channels in identified subtypes of neurons.
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Department of Physiology, University of Colorado, Health Sciences Center, Denver 80262, USA.
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Taylor
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D000818:Animals; D051662:Kv1.1 Potassium Channel; D008564:Membrane Potentials; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000737:chemistry; D009474:Neurons / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000096:biosynthesis* / Q000235:genetics; D024642:Potassium Channels, Voltage-Gated; D012333:RNA, Messenger / Q000096:biosynthesis; D014981:Xenopus
D009419:Nerve Tissue Proteins; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D012333:RNA, Messenger; D051662:Kv1.1 Potassium Channel