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Although acid-sensing ion channels (ASICs) are widely expressed in the central nervous system, their distribution and roles in axonal growth cones remain unclear. In this study, we examined ASIC localization and function in the axonal growth cones of cultured immature hippocampal neurons. Our immunocytochemical data showed that native and overexpressed ASIC1a and ASIC2a are both localized in growth cones of cultured young hippocampal neurons. Calcium imaging and electrophysiological assay results were utilized to validate their function. The calcium imaging test results indicated that the ASICs (primarily ASIC1a) present in growth cones mediate calcium influx despite the addition of voltage-gated Ca2+ channels antagonists and the depletion of intracellular calcium stores. The electrophysiological tests results suggested that a rapid decrease in extracellular pH at the growth cones of voltage-clamped neurons elicits inward currents that were blocked by bath application of the ASIC antagonist amiloride, showing that the ASICs expressed at growth cones are functional. The subsequent immuno-colocalization test results demonstrated that ASIC1a and ASIC2a are both colocalized with Neurofilament-H and Bassoon in mature hippocampal neurons. This finding demonstrated that after reaching maturity, ASIC1a and ASIC2a are both distributed in axons and the presynaptic membrane. Our data reveal the distribution of functional ASICs in growth cones of immature hippocampal neurons and the presence of ASICs in the axons and presynaptic membrane of mature hippocampal neurons, indicating a possible role for ASICs in axonal guidance, synapse formation and neurotransmitter release.

Figure 1. Transduced ASIC1a and ASIC2a is expressed at the axonal growth cones.

The study is aimed to explore the distribution and roles of acid-sensing ion channels (ASICs) in axonal growth cones. In this study, the authors examined ASIC localization and function in the axonal growth cones of cultured immature hippocampal neurons. They demonstrated that the ASIC1a and ASIC2a subunits are present in axonal growth cones from 3 days in vitro (DIV3) to DIV6 hippocampal neurons. They demonstrate that these ASICs are functional in the growth cones and are also expressed in the axons and presynaptic active zones of mature (DIV14) neurons, suggesting that they may regulate neurotransmitter release.

 
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