Giant excised patch recordings of recombinant ion channel currents expressed in mammalian cells.
The giant excised patch variant of patch clamp recording combines microsecond time resolution of macroscopic currents with rapid exchange of the experimental solutions at the intracellular membrane surface. This technique has been applied to a limited number of cell types, including Xenopus oocytes, muscle cells, and photoreceptors. We have applied this technique to recording recombinant ion channel currents expressed in membrane patches excised from HEK293 cell lines. Giant inside-out patch recordings of Na(+) channels and SK(Ca) type calcium-activated potassium channels show high temporal resolution and excellent signal to noise characteristics. This technique will facilitate the study of recombinant ion channels expressed in mammalian cells.
Couey
Jonathan J
JJ
Department of Anesthesia and Critical Care, University of Chicago, 5841 S. Maryland Avenue, Box 4028, Chicago, IL 60637, USA.
Ryan
Devon P
DP
Glover
John T
JT
Dreixler
John C
JC
Young
Joseph B
JB
Houamed
Khaled M
KM
D000818:Animals; D016477:Artifacts; D002460:Cell Line; D006801:Humans; D007473:Ion Channels / Q000502:physiology*; D008274:Magnesium / Q000494:pharmacology; D008322:Mammals; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000502:physiology; D009865:Oocytes / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods*; D020419:Photoreceptor Cells, Vertebrate / Q000502:physiology; D015221:Potassium Channels / Q000502:physiology; D024681:Potassium Channels, Calcium-Activated / Q000502:physiology; D011994:Recombinant Proteins; D051640:Small-Conductance Calcium-Activated Potassium Channels; D015222:Sodium Channels / Q000502:physiology; D014981:Xenopus
D007473:Ion Channels; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D011994:Recombinant Proteins; D051640:Small-Conductance Calcium-Activated Potassium Channels; D015222:Sodium Channels; D008274:Magnesium