Modulation of GABA flux across rat brain membranes resolved by a rapid quenched incubation technique

J. Kardos, Ilona Kovács, Tamás Blandl, Derek J. Cash

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The progress and inhibition of [3H]GABA influx in native plasma membrane vesicles from the rat cerebral cortex was studied on a subsecond to minute time scale under different conditions by applying a rapid quenched incubation technique. In the absence of Ca2+ ([Ca2+]free = 10-8 M), the progress of influx followed by the addition of 10 nM [3H]GABA to the membrane vesicle suspension with time (500 ms to 15 min) can be described by a first-order rate equation giving an overall rate constant, k, of 3.93 ± 0.48 × 10-3 s-1 and equilibrium influx value, INFe, of 8.84 ± 0.41 pmol [3H]GABA/mg protein. In the presence of Ca2+ ([Ca2+]free = 2.4 × 10-3 M) a significant increase in the INFe value was observed (k = 4.64 ± 0.41 × 10-3 s-1 and INFe = 13.9 ± 0.40 pmol[3H]GABA/mg protein). Multiplicity of GABA transporters was indicated in the time-dependent inhibition of [3H]GABA influx by different uptake blockers. In the absence of Ca2+, depolarization (75 mM KCl) inhibited the influx of [3H]GABA into the vesicles by ∼70% and initiated the efflux from vesicles loaded with [3H]GABA. Different uptake blockers inhibited the Ca2+-independent translocation of [3H]GABA in both directions with similar specificities.

Original languageEnglish
Pages (from-to)73-76
Number of pages4
JournalNeuroscience Letters
Volume182
Issue number1
DOIs
Publication statusPublished - Nov 21 1994

Fingerprint

gamma-Aminobutyric Acid
Membranes
Brain
GABA Plasma Membrane Transport Proteins
Cerebral Cortex
Suspensions
Proteins
Cell Membrane

Keywords

  • Membrane vesicle
  • Rapid quenched incubation technique
  • Rat cerebral cortex
  • [H]GABA translocation

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Modulation of GABA flux across rat brain membranes resolved by a rapid quenched incubation technique. / Kardos, J.; Kovács, Ilona; Blandl, Tamás; Cash, Derek J.

In: Neuroscience Letters, Vol. 182, No. 1, 21.11.1994, p. 73-76.

Research output: Contribution to journalArticle

Kardos, J. ; Kovács, Ilona ; Blandl, Tamás ; Cash, Derek J. / Modulation of GABA flux across rat brain membranes resolved by a rapid quenched incubation technique. In: Neuroscience Letters. 1994 ; Vol. 182, No. 1. pp. 73-76.
@article{0f530e249ebc407eaaf179d7bf18bcff,
title = "Modulation of GABA flux across rat brain membranes resolved by a rapid quenched incubation technique",
abstract = "The progress and inhibition of [3H]GABA influx in native plasma membrane vesicles from the rat cerebral cortex was studied on a subsecond to minute time scale under different conditions by applying a rapid quenched incubation technique. In the absence of Ca2+ ([Ca2+]free = 10-8 M), the progress of influx followed by the addition of 10 nM [3H]GABA to the membrane vesicle suspension with time (500 ms to 15 min) can be described by a first-order rate equation giving an overall rate constant, k, of 3.93 ± 0.48 × 10-3 s-1 and equilibrium influx value, INFe, of 8.84 ± 0.41 pmol [3H]GABA/mg protein. In the presence of Ca2+ ([Ca2+]free = 2.4 × 10-3 M) a significant increase in the INFe value was observed (k = 4.64 ± 0.41 × 10-3 s-1 and INFe = 13.9 ± 0.40 pmol[3H]GABA/mg protein). Multiplicity of GABA transporters was indicated in the time-dependent inhibition of [3H]GABA influx by different uptake blockers. In the absence of Ca2+, depolarization (75 mM KCl) inhibited the influx of [3H]GABA into the vesicles by ∼70{\%} and initiated the efflux from vesicles loaded with [3H]GABA. Different uptake blockers inhibited the Ca2+-independent translocation of [3H]GABA in both directions with similar specificities.",
keywords = "Membrane vesicle, Rapid quenched incubation technique, Rat cerebral cortex, [H]GABA translocation",
author = "J. Kardos and Ilona Kov{\'a}cs and Tam{\'a}s Blandl and Cash, {Derek J.}",
year = "1994",
month = "11",
day = "21",
doi = "10.1016/0304-3940(94)90209-7",
language = "English",
volume = "182",
pages = "73--76",
journal = "Neuroscience Letters",
issn = "0304-3940",
publisher = "Elsevier Ireland Ltd",
number = "1",

}

TY - JOUR

T1 - Modulation of GABA flux across rat brain membranes resolved by a rapid quenched incubation technique

AU - Kardos, J.

AU - Kovács, Ilona

AU - Blandl, Tamás

AU - Cash, Derek J.

PY - 1994/11/21

Y1 - 1994/11/21

N2 - The progress and inhibition of [3H]GABA influx in native plasma membrane vesicles from the rat cerebral cortex was studied on a subsecond to minute time scale under different conditions by applying a rapid quenched incubation technique. In the absence of Ca2+ ([Ca2+]free = 10-8 M), the progress of influx followed by the addition of 10 nM [3H]GABA to the membrane vesicle suspension with time (500 ms to 15 min) can be described by a first-order rate equation giving an overall rate constant, k, of 3.93 ± 0.48 × 10-3 s-1 and equilibrium influx value, INFe, of 8.84 ± 0.41 pmol [3H]GABA/mg protein. In the presence of Ca2+ ([Ca2+]free = 2.4 × 10-3 M) a significant increase in the INFe value was observed (k = 4.64 ± 0.41 × 10-3 s-1 and INFe = 13.9 ± 0.40 pmol[3H]GABA/mg protein). Multiplicity of GABA transporters was indicated in the time-dependent inhibition of [3H]GABA influx by different uptake blockers. In the absence of Ca2+, depolarization (75 mM KCl) inhibited the influx of [3H]GABA into the vesicles by ∼70% and initiated the efflux from vesicles loaded with [3H]GABA. Different uptake blockers inhibited the Ca2+-independent translocation of [3H]GABA in both directions with similar specificities.

AB - The progress and inhibition of [3H]GABA influx in native plasma membrane vesicles from the rat cerebral cortex was studied on a subsecond to minute time scale under different conditions by applying a rapid quenched incubation technique. In the absence of Ca2+ ([Ca2+]free = 10-8 M), the progress of influx followed by the addition of 10 nM [3H]GABA to the membrane vesicle suspension with time (500 ms to 15 min) can be described by a first-order rate equation giving an overall rate constant, k, of 3.93 ± 0.48 × 10-3 s-1 and equilibrium influx value, INFe, of 8.84 ± 0.41 pmol [3H]GABA/mg protein. In the presence of Ca2+ ([Ca2+]free = 2.4 × 10-3 M) a significant increase in the INFe value was observed (k = 4.64 ± 0.41 × 10-3 s-1 and INFe = 13.9 ± 0.40 pmol[3H]GABA/mg protein). Multiplicity of GABA transporters was indicated in the time-dependent inhibition of [3H]GABA influx by different uptake blockers. In the absence of Ca2+, depolarization (75 mM KCl) inhibited the influx of [3H]GABA into the vesicles by ∼70% and initiated the efflux from vesicles loaded with [3H]GABA. Different uptake blockers inhibited the Ca2+-independent translocation of [3H]GABA in both directions with similar specificities.

KW - Membrane vesicle

KW - Rapid quenched incubation technique

KW - Rat cerebral cortex

KW - [H]GABA translocation

UR - http://www.scopus.com/inward/record.url?scp=0028029588&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028029588&partnerID=8YFLogxK

U2 - 10.1016/0304-3940(94)90209-7

DO - 10.1016/0304-3940(94)90209-7

M3 - Article

C2 - 7891893

AN - SCOPUS:0028029588

VL - 182

SP - 73

EP - 76

JO - Neuroscience Letters

JF - Neuroscience Letters

SN - 0304-3940

IS - 1

ER -