T‐butyl‐bicyclo‐phosphorothionate (TBPS) is a prototypical representative of the cage‐convulsants which act through a use‐dependent block of the GABAA‐receptor‐ionophore complex. Using current recordings from cultured neurones of rat striatum the manner was investigated in which two antagonists, bicuculline and penicillin, presumably acting at the agonist binding site and in the ionic channel, respectively, modify the rate of block by TBPS.
Penicillin (5 or 10 mM) did not slow the rate of block by TBPS, but produced a significant enhancement of block rate, which, however, was inversely related to the degree of antagonism by penicillin of the GABA‐induced current.
Bicuculline (10 μM) reduced the rate of block by TBPS. However, this effect was 3 fold weaker than its GABA‐antagonistic action. The slowing of block rate and the current antagonism exhibited a biphasic, positive‐negative relationship. Co‐application of bicuculline (100 μM) in a concentration that produced nearly complete antagonism and TBPS (10 μM) resulted in a marked (∼40%) reduction of subsequent GABA response amplitudes compatible with a direct, bicuculline‐induced conformational change in the receptor required for the binding of and block by TBPS.
The lack of protection afforded by the channel blocker penicillin as well as the lack of correlation between bicuculline antagonism of the Cl−‐current and its efficiency in protecting against TBPS block is evidence against an open channel blocking mechanism for TBPS. TBPS does, therefore, not appear to gain access to its binding site via the open pore but through alternative routes regulated from the agonist binding site.
T‐butyl‐bicyclo‐phosphorothionate (TBPS) is a prototypical representative of the cage‐convulsants which act through a use‐dependent block of the GABAA‐receptor‐ionophore complex. Using current recordings from cultured neurones of rat striatum the manner was investigated in which two antagonists, bicuculline and penicillin, presumably acting at the agonist binding site and in the ionic channel, respectively, modify the rate of block by TBPS.
Penicillin (5 or 10 mM) did not slow the rate of block by TBPS, but produced a significant enhancement of block rate, which, however, was inversely related to the degree of antagonism by penicillin of the GABA‐induced current.
Bicuculline (10 μM) reduced the rate of block by TBPS. However, this effect was 3 fold weaker than its GABA‐antagonistic action. The slowing of block rate and the current antagonism exhibited a biphasic, positive‐negative relationship. Co‐application of bicuculline (100 μM) in a concentration that produced nearly complete antagonism and TBPS (10 μM) resulted in a marked (∼40%) reduction of subsequent GABA response amplitudes compatible with a direct, bicuculline‐induced conformational change in the receptor required for the binding of and block by TBPS.
The lack of protection afforded by the channel blocker penicillin as well as the lack of correlation between bicuculline antagonism of the Cl−‐current and its efficiency in protecting against TBPS block is evidence against an open channel blocking mechanism for TBPS. TBPS does, therefore, not appear to gain access to its binding site via the open pore but through alternative routes regulated from the agonist binding site.
British Journal of Pharmacology (2000) 129, 402–408; doi:10.1038/sj.bjp.0703063
DOI: 10.1038/sj.bjp.0703063
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