H 2 O 2 augments cytosolic calcium in nucleus tractus solitarii neurons via multiple voltage-gated calcium channels.

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 (H2 O2 ) 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 H2 O2 on cytosolic Ca2+ (via fura-2 imaging) and voltage-dependent calcium currents in dissociated rat nTS neurons. H2 O2 perfusion (200 µM; 1 min) induced a delayed, slow, and moderate increase (~27%) in intracellular Ca2+ concentration ([Ca2+ ]i ). The H2 O2 -mediated increase in [Ca2+ ]i prevailed during thapsigargin, excluding the endoplasmic reticulum as a Ca2+ source. The effect, however, was abolished by removal of extracellular Ca2+ or the addition of cadmium to the bath solution, suggesting voltage-gated Ca2+ channels (VGCCs) as targets for H2 O2 modulation. Recording of the total voltage-dependent Ca2+ current confirmed H2 O2 enhanced Ca2+ entry. Blocking VGCC L, N, and P/Q subtypes decreased the number of cells and their calcium currents that respond to H2 O2 The number of responder cells to H2 O2 also decreased in the presence of dithiothreitol, suggesting the actions of H2 O2 were dependent on sulfhydryl oxidation. In summary, here, we have shown that H2 O2 increases [Ca2+ ]i and its Ca2+ 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.