The role of sulfhydryl groups and calcium in the mercuric chloride-induced inhibition of glutamate uptake in rat primary astrocyte cultures

Jan Albrecht, Margaret Talbot, Harold K. Kimelberg, Michael Aschner

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Inhibition by mercuric chloride (MC) of the astrocytic uptake of the excitotoxic neurotransmitter L-glutamate (L-GLU) has been postulated to contribute to MC neurotoxicity. In the present study, we analyzed the ability of two sulfhydryl (SH)-protecting agents: a cell membrane non-penetrating compound-reduced glutathione (GSH), and the membrane permeable dithiothreitol (DTT), to reverse the inhibitory action of MC on the initial rate of uptake of radiolabelled GLU (100 μM) in primary cultures of rat astrocytes. MC at 5 μM concentration reduced the uptake to 46% of control when present in the incubation medium during the 5 min of actual uptake, and to 27% of control when astrocytes were preincubation with MC was partly relieved by the addition of 1 mM DTT during the actual 5 min uptake period. However, this inhibition could not be reversed by 1 mM GSH. Accordingly, it is postulated that the inhibitory effect exerted by MC on GLU uptake is associated with vulnerable SH groups located within, but not on the surface of the cell membrane. Neither 5 μM N-ethylmaleimide (NEM) nor 5 μM or 25 μM iodoacetate (IA) affected GLU uptake, indicating steric hindrance of the access of these two sulfhydryl reagents to the SH groups critical for the uptake. The effect of MC on GLU uptake was not altered by omission of Ca2+ or addition of a non-specific calcium channel blocker La3+ (10 μM), suggesting that the effect was not subsequent to Hg2+ entry into the cell through Ca2+ channels.

Original languageEnglish (US)
Pages (from-to)249-254
Number of pages6
JournalBrain Research
Volume607
Issue number1-2
DOIs
StatePublished - Apr 2 1993
Externally publishedYes

Fingerprint

Mercuric Chloride
Astrocytes
Glutamic Acid
Calcium
Dithiothreitol
Cell Membrane
Iodoacetates
Sulfhydryl Reagents
Ethylmaleimide
Calcium Channel Blockers
Glutathione
Neurotransmitter Agents
Membranes

Keywords

  • Astrocyte
  • Glutamate
  • Iodicetate
  • Mercuric chloride
  • N-Ethylmaleimide
  • Transport

ASJC Scopus subject areas

  • Developmental Biology
  • Molecular Biology
  • Clinical Neurology
  • Neuroscience(all)

Cite this

The role of sulfhydryl groups and calcium in the mercuric chloride-induced inhibition of glutamate uptake in rat primary astrocyte cultures. / Albrecht, Jan; Talbot, Margaret; Kimelberg, Harold K.; Aschner, Michael.

In: Brain Research, Vol. 607, No. 1-2, 02.04.1993, p. 249-254.

Research output: Contribution to journalArticle

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AB - Inhibition by mercuric chloride (MC) of the astrocytic uptake of the excitotoxic neurotransmitter L-glutamate (L-GLU) has been postulated to contribute to MC neurotoxicity. In the present study, we analyzed the ability of two sulfhydryl (SH)-protecting agents: a cell membrane non-penetrating compound-reduced glutathione (GSH), and the membrane permeable dithiothreitol (DTT), to reverse the inhibitory action of MC on the initial rate of uptake of radiolabelled GLU (100 μM) in primary cultures of rat astrocytes. MC at 5 μM concentration reduced the uptake to 46% of control when present in the incubation medium during the 5 min of actual uptake, and to 27% of control when astrocytes were preincubation with MC was partly relieved by the addition of 1 mM DTT during the actual 5 min uptake period. However, this inhibition could not be reversed by 1 mM GSH. Accordingly, it is postulated that the inhibitory effect exerted by MC on GLU uptake is associated with vulnerable SH groups located within, but not on the surface of the cell membrane. Neither 5 μM N-ethylmaleimide (NEM) nor 5 μM or 25 μM iodoacetate (IA) affected GLU uptake, indicating steric hindrance of the access of these two sulfhydryl reagents to the SH groups critical for the uptake. The effect of MC on GLU uptake was not altered by omission of Ca2+ or addition of a non-specific calcium channel blocker La3+ (10 μM), suggesting that the effect was not subsequent to Hg2+ entry into the cell through Ca2+ channels.

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