Na+ currents in cultured mouse pancreatic B-cells

Pancreatic B-cells, kept in culture for 1–4 days, were studied in the whole-cell, cell-attached and outside-out modes of the patch clamp technique. B-cells were identified by the appearance of electrical activity in the cell-attached mode when the bath glucose was raised from 3 to 20 mM. In whole-cell, 80% of these cells showed a transient inward Na+ current, when depolarizing pulses were preceded by holding potentials, or prepulses to potentials more negative than −80 mV. The midpoint (Eh) of the inactivation curve (h∞) was at −109 mV in 2.6 mM Ca2+, 1.2 mM Mg2+ and −120 mV in 0.2 mM Ca2+, 3.6 mM Mg2+. In 2.6 mM Ca2+, inactivation was fully removed atE<−150 mV. Na+ currents activated atE>−60 mV and were largest at around −10 mV (120 mM Na+). The kinetic parameters of activation (tp) and inactivation (τ)h were similar to those of other mammalian Na+ channels. Unitary currents with an amplitude of approximately 1 pA at −30 mV (140 mM Na+) with a similar voltage-dependence and time-course of mean current were recorded from outside-out patches. The results show that B-cells have a voltage-dependent Na+ current which, owing to the voltage-dependence of inactivation, is unlikely to play a major role in glucose-induced electrical activity.

[1]  Alain Marty,et al.  Tight-Seal Whole-Cell Recording , 1983 .

[2]  J. M. Fernández,et al.  Membrane patches and whole‐cell membranes: a comparison of electrical properties in rat clonal pituitary (GH3) cells. , 1984, The Journal of physiology.

[3]  P. Berggren,et al.  Voltage-activated Na+ currents and their suppression by phorbol ester in clonal insulin-producing RINm5F cells. , 1986, The American journal of physiology.

[4]  C. Starmer,et al.  Unitary sodium channels in isolated cardiac myocytes of rabbit. , 1983, Circulation research.

[5]  M. Cahalan,et al.  Chemical modification of sodium channel surface charges in frog skeletal muscle by trinitrobenzene sulphonic acid. , 1981, The Journal of physiology.

[6]  H. Lux,et al.  A low voltage-activated, fully inactivating Ca channel in vertebrate sensory neurones , 1984, Nature.

[7]  T. Narahashi,et al.  Isolation and kinetic analysis of inward currents in neuroblastoma cells , 1984, Neuroscience.

[8]  B. Lindemann,et al.  Patch-clamp study of isolated taste receptor cells of the frog , 2005, The Journal of Membrane Biology.

[9]  D Hof,et al.  A pulse generating and data recording system based on the microcomputer PDP 11/23. , 1986, Computer methods and programs in biomedicine.

[10]  P. Rorsman,et al.  Calcium and delayed potassium currents in mouse pancreatic beta‐cells under voltage‐clamp conditions. , 1986, The Journal of physiology.

[11]  J. M. Ritchie,et al.  Voltage-dependent sodium and potassium channels in mammalian cultured Schwann cells. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[12]  B. Sakmann,et al.  Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches , 1981, Pflügers Archiv.

[13]  D. Lowe,et al.  Correlations between chanelled Na+ entry, Ca2 + fluxes and insulin release in pancreatic beta-cells. , 1980, Hormone and metabolic research. Supplement series.

[14]  E Neher,et al.  A patch‐clamp study of bovine chromaffin cells and of their sensitivity to acetylcholine. , 1982, The Journal of physiology.

[15]  M. Rack,et al.  Effects of some chemical reagents on sodium current inactivation in myelinated nerve fibers of the frog. , 1986, Biophysical journal.

[16]  C. Pace,et al.  Glucose-induced electrical activity in the pancreatic beta-cell: effect of veratridine. , 1981, The American journal of physiology.

[17]  E Neher,et al.  Sodium and calcium channels in bovine chromaffin cells , 1982, The Journal of physiology.

[18]  H. Meves,et al.  The effect of temperature on the asymmetrical charge movement in squid giant axons. , 1979, The Journal of physiology.

[19]  P. Rorsman,et al.  Glucose dependent K+-channels in pancreaticβ-cells are regulated by intracellular ATP , 1985, Pflügers Archiv.

[20]  T. Kiss,et al.  Single Na channels in mouse neuroblastoma cell membrane , 1983, Pflügers Archiv.

[21]  H. Meissner,et al.  Significance of ionic fluxes and changes in membrane potential for stimulus-secretion coupling in pancreatic B-cells , 1984, Experientia.

[22]  R. Tsien,et al.  A novel type of cardiac calcium channel in ventricular cells. , 1985, Nature.

[23]  H Reuter,et al.  Sodium channels in cultured cardiac cells. , 1983, The Journal of physiology.

[24]  H. Lux,et al.  Sodium channels in cultured chick dorsal root ganglion neurons , 1986, European Biophysics Journal.

[25]  R. Tsien,et al.  Three types of neuronal calcium channel with different calcium agonist sensitivity , 1985, Nature.

[26]  L. Satin,et al.  Voltage-gated Ca current in pancreatic islet beta-cells. , 1986, Advances in experimental medicine and biology.

[27]  H. Meissner,et al.  Ionic mechanisms of the glucose-induced membrane potential changes in B-cells. , 1980, Hormone and metabolic research. Supplement series.

[28]  Å. Lernmark,et al.  The preparation of, and studies on, free cell suspensions from mouse pancreatic islets , 1974, Diabetologia.

[29]  Stephen J. H. Ashcroft,et al.  Glucose induces closure of single potassium channels in isolated rat pancreatic β-cells , 1984, Nature.

[30]  D L Kunze,et al.  Cardiac Na currents and the inactivating, reopening, and waiting properties of single cardiac Na channels , 1985, The Journal of general physiology.

[31]  L. Satin,et al.  Voltage-gated Ca2+ current in pancreatic B-cells , 1985, Pflügers Archiv.

[32]  O. Petersen,et al.  Electrophysiology of the pancreas. , 1987, Physiological reviews.