Reactive oxygen species (ROS) play a profound role in cardiorespiratory function under normal physiological conditions and disease states. ROS can influence neuronal activity by altering various ion channels and transporters. Within the nucleus tractus solitarii (nTS), a vital brainstem area for cardiorespiratory control, hydrogen peroxide (H2O2) induces sustained hyperexcitability following an initial depression of neuronal activity. The mechanism(s) associated with the delayed hyperexcitability are unknown. Here we evaluate the effect(s) of H2O2 on cytosolic Ca++ (via Fura-2 imaging) and voltage-dependent calcium currents in dissociated rat nTS neurons. H2O2 perfusion (200 µM; 1 min) induced a delayed, slow, and moderate increase (~27%) in intracellular Ca++ ([Ca++]i). The H2O2-mediated increase in [Ca++]i prevailed during Thapsigargin, excluding the endoplasmic reticulum as a Ca++ source. The effect, however, was abolished by removal of extracellular Ca++ or the addition of cadmium to the bath solution, suggesting voltage-gated Ca++ channels (VGCCs) as targets for H2O2 modulation. Recording of the total voltage-dependent Ca++ current confirmed H2O2 enhanced Ca++ entry. Blocking VGCC L-, N-, and P/Q-subtype decreased the number of cells and their calcium currents that respond to H2O2. The number of responder cells to H2O2 also decreased in the presence of Dithiothreitol, suggesting the actions of H2O2 were dependent on sulfhydryl-oxidation. In summary, here, we have shown that H2O2 increases [Ca++]i and their Ca++ currents, which is dependent on multiple VGCCs likely by oxidation of sulfhydryl groups. These processes presumably contribute to the previously observed delayed hyperexcitability of nTS neurons in in vitro brainstem slices.
- autonomic nervous system
- redox signaling
- voltage-gated calcium channel
- Copyright © 2017, American Journal of Physiology-Cell Physiology