关键词:
voltage-activated calcium channels
hypoxia
rat embryo chromaffin cells
摘要:
At early life, the adrenal chromaffin cells respond with a catecholamine surge under hypoxic conditions. This response depends on Ca2+ entry through voltage-activated calcium channels (VACCs). We have investigated here three unresolved questions that concern this response in rat embryo chromaffin cells (ECCs): 1) the relative contribution of L (alpha(1D), Cav1.3), N (alpha(1B), Cav2.2), and PQ (alpha(1A), Cav2.1) to the whole cell Ca2+ current (I-Ca);2) the relative contribution of L and N/PQ channels to the cytosolic Ca2+ elevations triggered by hypoxia (Delta[Ca2+](c));and 3) the role of L and non-L high-VACCs in the regulation of the catecholamine surge occurring during prolonged (1 min) hypoxia exposure of ECCs. Nimodipine halved peak I-Ca and blocked 60% the total Ca2+ entry during a 50-ms depolarizing pulse to 0 mV (Q(Ca)). Combined omega-agatoxin IVA plus omega-conotoxin GVIA (Aga/GVIA) blocked 30% of both I-Ca peak and Q(Ca). This relative proportion of L- and non-L VACCs was corroborated by Western blot that indicated 55, 23, and 25% relative expression of L, N, and PQ VACCs. Exposure of ECCs to hypoxia elicited a mild but sustained Delta[Ca2+](c);the area of Delta[Ca2+](c) was blocked 50% by nifedipine and 10% by Aga/GVIA. Exposure of ECCs to 1-min hypoxia elicited an initial transient burst of amperometric secretory spikes followed by scattered spikes along the time of cell exposure to hypoxia. This bulk response was blocked 85% by nimodipine and 35% by Aga/GVIA. Histograms on secretory spike frequency vs. time indicated a faster initial inactivation when Ca2+ entry took place through N/PQ channels;more sustained secretion but at a lower rate was associated to Ca2+ entry through L channels. The results suggest that the HIS response may initially be controlled by L and P/Q channels, but later on, N/PQ channels inactivate and the delayed HIS response is maintained at lower rate by slow-inactivating L channels.