Voltage-dependent calcium and potassium channels in Schwann cells cultured from dorsal root ganglia of the mouse
1. Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18-19 days). 2. In standard external solution, from a holding potential of -70 mV, two types of voltage-dependent K+ currents were recorded: a fast transien...
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Published in | The Journal of physiology Vol. 441; no. 1; pp. 35 - 56 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Oxford
The Physiological Society
01.09.1991
Blackwell |
Subjects | |
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Abstract | 1. Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from
OF1 mice embryos (18-19 days). 2. In standard external solution, from a holding potential of -70 mV, two types of voltage-dependent
K+ currents were recorded: a fast transient current and a delayed sustained current. With a holding potential of -30 mV, only
the delayed sustained current could be evoked. 3. Both K+ currents were inhibited by tetraethylammonium chloride (TEA) and
4-aminopyridine (4-AP) in a dose-dependent manner. For the transient current the half-maximal effective dose was 100 mM for
TEA and 1.3 mM for 4-AP. For the delayed sustained current the half-maximal effective dose was 11 mM for TEA and 4 mM for
4-AP. Both currents were insensitive to external Ca2+. 4. The delayed sustained current, isolated by use of a holding potential
of -30 mV displayed a 'cumulative inactivation' which was removed by hyperpolarizing the membrane to -70 mV between each test
pulse. 5. In K(+)-free external and pipette solutions, with 10 mM-external Ca2+, from a holding potential of -70 mV voltage-dependent
Ca2+ channel currents were recorded. The threshold for activation was -45.3 +/- 5.4 mV (mean +/- S.D., n = 5) and the current
inactivated fully at the end of the test potential. The current was unaffected by 2 microM-tetrodotoxin (TTX) and totally
blocked by 5 mM-Co2+. 6. Equimolar replacement of external Ca2+ by Ba2+ did not significantly modify the voltage dependence
(threshold for activation -42.8 +/- 6.4 mV, n = 7) or the magnitude of the inward current. Ca2+ and Ba2+ were equally permeant.
The fully inactivating current was insensitive to both nifedipine and Bay K 8644 (1 microM each). Increasing the external
Ba2+ concentration from 10 to 89 mM enhanced the Ba2+ current and shifted the voltage dependence of the current (threshold
for activation, -30.5 +/- 7.3 mV, n = 9) along the voltage axis as expected for altered external surface potential. 7. In
89 mM-external Ba2+ solution, some cells displayed an additional slowly decaying current which was totally blocked by nifedipine
(1 microM). 8. Ca2+ channel currents were recorded only when DRG neurons were present in the culture, as excision of explants
and subsequent axonal degeneration led to loss of detectable Ca2+ channel currents. This phenomenon was never observed for
K+ currents. 9. We conclude that mouse Schwann cells in organotypic culture possess voltage-dependent K+ and Ca2+ channels. |
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AbstractList | 1. Whole‐cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18‐19 days). 2. In standard external solution, from a holding potential of ‐70 mV, two types of voltage‐dependent K+ currents were recorded: a fast transient current and a delayed sustained current. With a holding potential of ‐30 mV, only the delayed sustained current could be evoked. 3. Both K+ currents were inhibited by tetraethylammonium chloride (TEA) and 4‐aminopyridine (4‐AP) in a dose‐dependent manner. For the transient current the half‐maximal effective dose was 100 mM for TEA and 1.3 mM for 4‐AP. For the delayed sustained current the half‐maximal effective dose was 11 mM for TEA and 4 mM for 4‐AP. Both currents were insensitive to external Ca2+. 4. The delayed sustained current, isolated by use of a holding potential of ‐30 mV displayed a ‘cumulative inactivation’ which was removed by hyperpolarizing the membrane to ‐70 mV between each test pulse. 5. In K(+)‐free external and pipette solutions, with 10 mM‐external Ca2+, from a holding potential of ‐70 mV voltage‐dependent Ca2+ channel currents were recorded. The threshold for activation was ‐45.3 +/‐ 5.4 mV (mean +/‐ S.D., n = 5) and the current inactivated fully at the end of the test potential. The current was unaffected by 2 microM‐tetrodotoxin (TTX) and totally blocked by 5 mM‐Co2+. 6. Equimolar replacement of external Ca2+ by Ba2+ did not significantly modify the voltage dependence (threshold for activation ‐42.8 +/‐ 6.4 mV, n = 7) or the magnitude of the inward current. Ca2+ and Ba2+ were equally permeant. The fully inactivating current was insensitive to both nifedipine and Bay K 8644 (1 microM each). Increasing the external Ba2+ concentration from 10 to 89 mM enhanced the Ba2+ current and shifted the voltage dependence of the current (threshold for activation, ‐30.5 +/‐ 7.3 mV, n = 9) along the voltage axis as expected for altered external surface potential. 7. In 89 mM‐external Ba2+ solution, some cells displayed an additional slowly decaying current which was totally blocked by nifedipine (1 microM). 8. Ca2+ channel currents were recorded only when DRG neurons were present in the culture, as excision of explants and subsequent axonal degeneration led to loss of detectable Ca2+ channel currents. This phenomenon was never observed for K+ currents. 9. We conclude that mouse Schwann cells in organotypic culture possess voltage‐dependent K+ and Ca2+ channels. Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18-19 days). In standard external solution, from a holding potential of -70 mV, two types of voltage-dependent K super(+) currents were recorded: a fast transient current and a delayed sustained current. Both K super(+) current were inhibited by tetraethylammonium chloride (TEA) and 4-aminopyridine (4-AP) in a dose-dependent manner. We conclude that mouse Schwann cells in organotypic culture possess voltage-dependent K super(+) and Ca super(2+) channels. The fast transient K super(+) current is similar to A currents described in numerous neuronal cells. The delayed sustained current is similar to the delayed rectifier K super(+) current widely distributed among excitable cells. Two types of Ca super(2+) channel currents are present on mouse Schwann cells: a fully inactivating current which resembles the T-type current more particularly described in neuronal cells and a slowly decaying current similar to the L-type current. 1. Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18-19 days). 2. In standard external solution, from a holding potential of -70 mV, two types of voltage-dependent K+ currents were recorded: a fast transient current and a delayed sustained current. With a holding potential of -30 mV, only the delayed sustained current could be evoked. 3. Both K+ currents were inhibited by tetraethylammonium chloride (TEA) and 4-aminopyridine (4-AP) in a dose-dependent manner. For the transient current the half-maximal effective dose was 100 mM for TEA and 1.3 mM for 4-AP. For the delayed sustained current the half-maximal effective dose was 11 mM for TEA and 4 mM for 4-AP. Both currents were insensitive to external Ca2+. 4. The delayed sustained current, isolated by use of a holding potential of -30 mV displayed a 'cumulative inactivation' which was removed by hyperpolarizing the membrane to -70 mV between each test pulse. 5. In K(+)-free external and pipette solutions, with 10 mM-external Ca2+, from a holding potential of -70 mV voltage-dependent Ca2+ channel currents were recorded. The threshold for activation was -45.3 +/- 5.4 mV (mean +/- S.D., n = 5) and the current inactivated fully at the end of the test potential. The current was unaffected by 2 microM-tetrodotoxin (TTX) and totally blocked by 5 mM-Co2+. 6. Equimolar replacement of external Ca2+ by Ba2+ did not significantly modify the voltage dependence (threshold for activation -42.8 +/- 6.4 mV, n = 7) or the magnitude of the inward current. Ca2+ and Ba2+ were equally permeant. The fully inactivating current was insensitive to both nifedipine and Bay K 8644 (1 microM each). Increasing the external Ba2+ concentration from 10 to 89 mM enhanced the Ba2+ current and shifted the voltage dependence of the current (threshold for activation, -30.5 +/- 7.3 mV, n = 9) along the voltage axis as expected for altered external surface potential. 7. In 89 mM-external Ba2+ solution, some cells displayed an additional slowly decaying current which was totally blocked by nifedipine (1 microM). 8. Ca2+ channel currents were recorded only when DRG neurons were present in the culture, as excision of explants and subsequent axonal degeneration led to loss of detectable Ca2+ channel currents. This phenomenon was never observed for K+ currents. 9. We conclude that mouse Schwann cells in organotypic culture possess voltage-dependent K+ and Ca2+ channels. |
Author | T Amédée B Dupouy E Ellie J D Vincent |
AuthorAffiliation | Institut National de la Santé et de la Recherche Médicale, Unité de Neurobiologie Intégrative, Bordeaux, France |
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Keywords | Peripheral nervous system Cell culture Calcium Rodentia Electrophysiology Patch clamp method Ionic channel Neuroglía Vertebrata Mammalia Mouse Schwann cell Spinal ganglion Membrane potential Potassium |
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Snippet | 1. Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from
OF1 mice embryos (18-19 days). 2.... 1. Whole‐cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18‐19 days). 2.... 1. Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18-19 days). 2.... Whole-cell patch clamp studies were carried out on Schwann cells in organotypic cultures of dorsal root ganglia (DRG) from OF1 mice embryos (18-19 days). In... |
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StartPage | 35 |
SubjectTerms | 4-Aminopyridine - pharmacology Animals Barium - metabolism Biological and medical sciences Calcium Channels - drug effects Calcium Channels - physiology Cells, Cultured Cobalt - pharmacology dorsal root ganglia Dose-Response Relationship, Drug Fundamental and applied biological sciences. Psychology Ganglia, Spinal - cytology Isolated neuron and nerve. Neuroglia Membrane Potentials - drug effects Mice Nifedipine - pharmacology potassium Potassium Channels - drug effects Potassium Channels - physiology Schwann cells Schwann Cells - physiology Tetraethylammonium Tetraethylammonium Compounds - pharmacology Tetrodotoxin - pharmacology Vertebrates: nervous system and sense organs |
Title | Voltage-dependent calcium and potassium channels in Schwann cells cultured from dorsal root ganglia of the mouse |
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