THE ROLE OF SULFHYDRYL-GROUPS AND CALCIUM IN THE MERCURIC CHLORIDE-INDUCED INHIBITION OF GLUTAMATE UPTAKE IN RAT PRIMARY ASTROCYTE CULTURES

被引:58
作者
ALBRECHT, J
TALBOT, M
KIMELBERG, HK
ASCHNER, M
机构
[1] ALBANY MED COLL, DEPT PHARMACOL & TOXICOL, A-136, 47 NEW SCOTLAND AVE, ALBANY, NY 12208 USA
[2] POLISH ACAD SCI, MED RES CTR, DEPT NEUROPATHOL, WARSAW 42, POLAND
[3] ALBANY MED COLL, DIV NEUROSURG, ALBANY, NY 12208 USA
关键词
MERCURIC CHLORIDE; N-ETHYLMALEIMIDE; IODOACETATE; GLUTAMATE; ASTROCYTE; TRANSPORT;
D O I
10.1016/0006-8993(93)91513-R
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
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 muM) in primary cultures of rat astrocytes. MC at 5 muM 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 preincubated for 30 min in HEPES buffer containing MC prior to GLU uptake measurements. GLU uptake inhibition caused by 30 min 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 muM N-ethylmaleimide (NEM) nor 5 muM or 25 muM 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 muM), suggesting that the effect was not subsequent to Hg2+ entry into the cell through Ca2+ channels.
引用
收藏
页码:249 / 254
页数:6
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