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—The effects of hypoxia on the L-type Ca2+ current (I Ca-L ) in the absence and presence of the β-adrenergic receptor agonist isoproterenol (Iso) were examined. Exposing guinea pig ventricular myocytes to hypoxia alone resulted in a reversible inhibition of basalI Ca-L . When cells were exposed to Iso in the presence of hypoxia, theK 0.5 for activation ofI Ca-L by Iso was significantly decreased from 5.3±0.7 to 1.6±0.1 nmol/L. The membrane-impermeant thiol-specific oxidizing compound 5,5′-dithio-bis(2-nitrobenzoic acid) (DTNB) attenuated the inhibition of basalI Ca-L by hypoxia 81.3±9.4% but had no effect on the increase in sensitivity ofI Ca-L to Iso. In addition, DTT mimicked the effects of hypoxia on basalI Ca-L and the increase in sensitivity to Iso. Neither the inhibitors of guanylate cyclase LY-83583 or methylene blue nor the NO synthase inhibitorN G -monomethyl-l -arginine monoacetate had any effect on the basal inhibition ofI Ca-L or the decrease inK 0.5 for activation ofI Ca-L by Iso during hypoxia. However, the protein kinase C (PKC) inhibitors bisindolylmaleimide I and Gö 7874 significantly attenuated the increase in sensitivity ofI Ca-L to Iso. More specifically, the response was attenuated when cells were dialyzed with a peptide inhibitor of the C2 region–containing classical PKC isoforms. The same effect was not observed with the PKCε peptide inhibitor. These results suggest that hypoxia regulatesI Ca-L through the following 2 distinct mechanisms: direct inhibition of basalI Ca-L and an indirect effect on the sensitivity of the channel to β-adrenergic receptor stimulation that is mediated through a classical PKC isoform.

circulation research

Hypoxia Increases the Sensitivity of the L-Type Ca <sup>2+</sup> Current to β-Adrenergic Receptor Stimulation via a C2 Region–Containing Protein Kinase C Isoform

Hypoxia Increases the Sensitivity of the L-Type Ca 2+ Current to β-Adrenergic Receptor Stimulation via a C2 Region–Containing Protein Kinase C Isoform

Hypoxia Increases the Sensitivity of the L-Type Ca ²⁺ Current to β-Adrenergic Receptor Stimulation via a C2 Region–Containing Protein Kinase C Isoform