Abstract
Chloride ion (Cl−) is one of the most abundant anions in our body. Increasing evidence suggests that Cl− plays fundamental roles in various cellular functions. We have previously reported that electroneutral cation-chloride cotransporters, such as Na+-K+-2Cl− cotransporter 1 (NKCC1) and K+-Cl− cotransporter 1 (KCC1), are involved in neurite outgrowth during neuronal differentiation. In the present study, we studied if there is correlation between intracellular Cl− concentrations ([Cl−]i) and the length of growing neurites. We measured [Cl−]i in the cell body and growing neurite tips using halide-sensitive fluorescent dye N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE), revealing that [Cl−]i in the tip of growing neurite was higher than that in cell body in a single cell. Importantly, there was a significant positive correlation between the length of growing neurite and [Cl−]i in neurite tip. Bumtanide (BMT), an inhibitor of NKCC1, significantly inhibited neurite outgrowth and decreased [Cl−]i in neurite tip. The results obtained in the present study and our previous studies together strongly suggest that high [Cl−]i in neurite tip region is crucial for efficient neurite outgrowth.
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