Abstract
We describe here the properties of calcium green-5N (CG5N), a novel Ca 2+-sensitive fluorescent probe. CG5N binds Ca 2+ with an affinity of 4.29 ± 0.67 μM. Ca 2+ binding is associated with a 14.7-fold increase in fluorescence. Mg 2+ binds to CG5N with an affinity of 5 mM but produces a very small change in fluorescence. Using CG5N [Ca 2+] i was recorded in single cultured neurons from fetal rat forebrain. Glutamate together with glycine increased [Ca 2+] i by 1.17 ± 0.15 μM. Pretreating cells with 5 mM potassium cyanide significantly potentiated the response to glutamate, resulting in a mean increase of 8.8 ± 2 μM. These results demonstrate that CG5N is a useful indicator for measuring very large changes in [Ca 2+] i in neurons.
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