Abstract
We recently used Western blots for connexin 36 and neuronal dye coupling with neurobiotin to measure developmental decrease in neuronal gap junction coupling in cell cultures. To ask whether Ca 2+ imaging also can be used to measure changes in the amount of neuronal gap junction coupling, we defined a Ca 2+ coupling coefficient as the percentage of neurons with bicuculline-induced increases in intracellular Ca 2+ that are suppressed by blocking gap junctions. We demonstrate in rat and mouse hypothalamic neuronal cultures that the Ca 2+ coupling coefficient decreases during culture development, this decrease is prevented by manipulations that also prevent developmental decrease in neuronal gap junction coupling, and the coefficient is low in cultures lacking connexin 36. The results indicate that Ca 2+ imaging is a useful tool to quantify the amount of neuronal gap junction coupling in cultures.
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