Calcium clamp in single nerve cells

P. V. Belan; P. G. Kostyuk; V. A. Snitsarev; A. V. Tepikin

doi:10.1016/0143-4160(93)90001-M

Calcium clamp in single nerve cells

P. V. Belan, P. G. Kostyuk, V. A. Snitsarev, A. V. Tepikin

Biology

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

Free calcium concentration in isolated single neurons was clamped using a new technical approach based on a feed-back connection between the Fura-2 fluorescence signal measuring the intracellular Ca²⁺ concentration ([Ca²⁺]_i) and iontophoretic current injecting Ca²⁺ into the cell. Beginning of [Ca²⁺]_i clamping at a level above the basal one triggered fast (few seconds) current transients equal to injection of 36 ± 20 μM Ca²⁺ (for a 0.1 μM change of [Ca²⁺]_i), representing the filling of a fast cytosolic buffer. Continuation of clamping required very small clamping currents (corresponding to injection of 0.39 ± 0.20 μM.s^-1 Ca²⁺). This value increased proportionally to the magnitude of the change of [Ca²⁺]_i above basal level, indicating the activation of calcium-dependent mechanisms for Ca²⁺ removal from the cytosol. The described approach allowed measurement, under physiological conditions, of the capacitative and kinetic properties of different Ca-regulating systems functioning in a single nerve cell as well as other types of cells.

Original language	English
Pages (from-to)	419-425
Number of pages	7
Journal	Cell Calcium
Volume	14
Issue number	6
DOIs	https://doi.org/10.1016/0143-4160(93)90001-M
State	Published - Jun 1993

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title = "Calcium clamp in single nerve cells",

abstract = "Free calcium concentration in isolated single neurons was clamped using a new technical approach based on a feed-back connection between the Fura-2 fluorescence signal measuring the intracellular Ca2+ concentration ([Ca2+]i) and iontophoretic current injecting Ca2+ into the cell. Beginning of [Ca2+]i clamping at a level above the basal one triggered fast (few seconds) current transients equal to injection of 36 ± 20 μM Ca2+ (for a 0.1 μM change of [Ca2+]i), representing the filling of a fast cytosolic buffer. Continuation of clamping required very small clamping currents (corresponding to injection of 0.39 ± 0.20 μM.s-1 Ca2+). This value increased proportionally to the magnitude of the change of [Ca2+]i above basal level, indicating the activation of calcium-dependent mechanisms for Ca2+ removal from the cytosol. The described approach allowed measurement, under physiological conditions, of the capacitative and kinetic properties of different Ca-regulating systems functioning in a single nerve cell as well as other types of cells.",

author = "Belan, {P. V.} and Kostyuk, {P. G.} and Snitsarev, {V. A.} and Tepikin, {A. V.}",

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T1 - Calcium clamp in single nerve cells

AU - Belan, P. V.

AU - Kostyuk, P. G.

AU - Snitsarev, V. A.

AU - Tepikin, A. V.

PY - 1993/6

Y1 - 1993/6

N2 - Free calcium concentration in isolated single neurons was clamped using a new technical approach based on a feed-back connection between the Fura-2 fluorescence signal measuring the intracellular Ca2+ concentration ([Ca2+]i) and iontophoretic current injecting Ca2+ into the cell. Beginning of [Ca2+]i clamping at a level above the basal one triggered fast (few seconds) current transients equal to injection of 36 ± 20 μM Ca2+ (for a 0.1 μM change of [Ca2+]i), representing the filling of a fast cytosolic buffer. Continuation of clamping required very small clamping currents (corresponding to injection of 0.39 ± 0.20 μM.s-1 Ca2+). This value increased proportionally to the magnitude of the change of [Ca2+]i above basal level, indicating the activation of calcium-dependent mechanisms for Ca2+ removal from the cytosol. The described approach allowed measurement, under physiological conditions, of the capacitative and kinetic properties of different Ca-regulating systems functioning in a single nerve cell as well as other types of cells.

AB - Free calcium concentration in isolated single neurons was clamped using a new technical approach based on a feed-back connection between the Fura-2 fluorescence signal measuring the intracellular Ca2+ concentration ([Ca2+]i) and iontophoretic current injecting Ca2+ into the cell. Beginning of [Ca2+]i clamping at a level above the basal one triggered fast (few seconds) current transients equal to injection of 36 ± 20 μM Ca2+ (for a 0.1 μM change of [Ca2+]i), representing the filling of a fast cytosolic buffer. Continuation of clamping required very small clamping currents (corresponding to injection of 0.39 ± 0.20 μM.s-1 Ca2+). This value increased proportionally to the magnitude of the change of [Ca2+]i above basal level, indicating the activation of calcium-dependent mechanisms for Ca2+ removal from the cytosol. The described approach allowed measurement, under physiological conditions, of the capacitative and kinetic properties of different Ca-regulating systems functioning in a single nerve cell as well as other types of cells.

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U2 - 10.1016/0143-4160(93)90001-M

DO - 10.1016/0143-4160(93)90001-M

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AN - SCOPUS:0027219591

SN - 0143-4160

VL - 14

SP - 419

EP - 425

JO - Cell Calcium

JF - Cell Calcium

IS - 6

ER -

Calcium clamp in single nerve cells

Abstract

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