Evidence for involvement of the PKC-α isoform in myogenic contractions of the coronary microcirculation

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Abstract
The role of protein kinase C (PKC) isoforms in myogenic tone of the ferret coronary microcirculation was investigated by measuring fura 2 Ca2+signals, PKC immunoblots, contractile responses, and confocal microscopy of PKC translocation. Phorbol ester-evoked contractions were completely abolished in the absence of extracellular Ca2+but involved a Ca2+sensitization relative to KCl contractions. Immunoblotting using isoform-specific antibodies showed the presence of PKC-α and -ι and traces of PKC-ɛ and -μ in the ferret coronary microcirculation. PKC-β was not detectable. When intraluminal pressure (40 to 60 and 80 mmHg) was increased, ferret coronary arterioles showed a transient increase in fura 2 Ca2+signals, whereas the myogenic tone remained sustained. The increase in Ca2+and tone was sustained at 100 mmHg. Isolated ferret coronary arterioles were fixed and immunostained for PKC-α at 40 and 100 mmHg intraluminal pressure. PKC translocation was determined by confocal microscopy. Increased PKC translocation was observed when vessels were exposed to 100 mmHg relative to that at resting pressure (40 mmHg). These results suggest a link between the Ca2+sensitization that occurs during the myogenic contraction and activation of the α-isoform of PKC.