Abstract:
The effects of AMG- 1 6 (5-hydroxy-2-methylpyridylamino)-9 ribofranosylpurine, an adenosine analogue, and adenosine on Ca
2+-dependent and independent release of glutamate from rat synaptosomes induced by KC1 30 mmol · L
-1 were studied with an enzyme-linked fluorometric assay. The synaptosomes were prepared and preincubated for 30 min at 37℃. Two ml of incubation mixture containing synaptosomes(1.27 mg protein), NADP
+ 1 mmol· L
-1,
L-glutamic dehydrogenase 50 U, CaCl
2 1.3 mmol ·L
-1 (or EGTA 1.3 mmol, L
-1) was transferred to the stirred cuvette in the fluorometer at 37℃ for 5 min. Then, AMG-1 (or adenosine) was added. Released glutamate (eg. fluorescent intensity) was monitored following the addition of 30 mmol · L
-1 KCl. The results indicate that Ca
2+-dependent glutamate release from depolarized synaptosomes is inhibited by AMG-1 (0.1 mmol · L
-1) or adenosine ( 0.3 mmol · L
-1). The action of AMG-1 seems to be similar to that of adenosine. However, no change was found on Ca
2+-independent release of glutamate. This implies that the protective effect of AMG-1 against cerebral ischemia may be partially due to inhibiting glutamate release from nerve terminal via the activation of adenosine A
1 receptor.