Characteristics of type I and type II K+ channels in rabbit cultured Schwann cells.
1. Voltage-dependent K+ currents were studied in rabbit Schwann cells cultured from neonatal sciatic nerve and from the lumbar or sacral spinal roots of 10-day-old animals. 2. Whole-cell K+ currents, evoked in response to depolarizing voltage-clamp steps, were categorized as type I or type II on the basis of their apparent threshold and activation kinetics. In the presence of a quasi-physiological [K+] gradient, the magnitude of the fully activated type I current varied linearly with membrane potential, whereas type II current always gave rise to a curved and outwardly rectifying current-membrane potential (I-E) relation. 3. Type II whole-cell currents, obtained with long duration voltage-clamp steps (> or = 1 s), have an apparent threshold for activation close to -40 mV. Type II current inactivated slowly, and apparently to completion. The current is more than 90% inactivated over 5 s at 0 mV (time consant of inactivation, tau h, approximately 2 s, 20-22 degrees C). Type I current, which activates at close to -60 mV, inactivated at about half this rate at the same potential, assuming that inactivation also proceeds to completion. 4. Type I whole-cell currents were reversibly blocked by superfused beta-bungarotoxin (beta-BuTX; apparent KD = 46 nM). beta-BuTX did not appear to reduce type II whole-cell currents at concentrations up to 500 nM. 5. In outside-out patches, the type I channel had an almost linear I-E relation over the potential range -60 to +60 mV with a quasi-physiological [K+] gradient. A best linear fit gave a single-channel conductance of 12 pS under these conditions. In symmetrical 170 mM K+, type I channels had a single-channel conductance of 30 pS over the same potential range. 6. More slowly activating type II single-channel currents were also recorded in inside-out patches. With symmetrical 170 mM K+, the major conductance level was close to 9.0 pS. With a quasi-physiological [K+] gradient, type II single channels exhibit outward rectification that is reasonably well described by the Goldman-Hodgkin-Katz current equation. 7. In the presence of 2 nM externally superfused alpha-dendrotoxin (alpha-DTX), or 50 nM superfused beta-BuTX, unitary currents were recorded (outside-out patches, -60 or -50 mV) that were smaller than control type I currents. Virtually all transitions in the presence of 50 nM beta-BuTX were at one-third of the control current level. The currents did not conform to the characteristics of type II. 8. The electrophysiological and pharmacological characteristics of the type I channel strongly suggest that it is a member of the mammalian K+ channel subfamily of Shaker homologues, most similar to the homomultimeric Kv1.1 translation product. The type II channel may be a member of the mammalian Shab subfamily. 9. Possible roles for Na+ channels and type I K+ channels in the Schwann cell are discussed.
Baker
M D
MD
Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520-8066, USA.
Ritchie
J M
JM
D000818:Animals; D002038:Bungarotoxins / Q000494:pharmacology; D002478:Cells, Cultured; D007700:Kinetics; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000502:physiology*; D011817:Rabbits; D012583:Schwann Cells / Q000502:physiology*; D012584:Sciatic Nerve / Q000502:physiology*; D013116:Spinal Cord / Q000502:physiology*
D002038:Bungarotoxins; D015221:Potassium Channels