Low frequency oscillations of murine skin microcirculations and periodic changes of [Ca2+]i and [NO]i levels in murine endotheliocytes: an effect of provocative tests

The five frequency intervals of skin blood oscillation were described: cardiac, respiratory, myogenic, neurogenic and endothelial ones. The endothelial interval is derived into NO-independent and NO-dependent. Exact molecular, cell or systemic mechanisms of endothelial oscillations generation are unclear. We proposed that oscillations of Ca2+ and NO in endotheliocytes may be possible source of skin blood perfusion (SBP) oscillations in endothelial interval. To examine our hypothesis we compared the oscillations of cytoplasmic Ca2+ and NO ([Ca2+]i and [NO]i) concentration in cultured murine microvascular endotheliocytes and SBP oscillations in mice. Local heating test and model hypoxia were used as tools to evaluate an interconnection of studied parameters. [Ca2+]i and [NO]i were measured simultaneously by Fura-2 AM and DAF-FM. The SBP was measured by laser Doppler flowmetry. The [Ca2+]i and [NO]i oscillations at 0.005-0.01 Hz were observed in endotheliocytes, that coincides the ranges of NO-independent endothelial interval. Heating decreased amplitude of [Ca2+]i and [NO]i oscillations in cells in NO-independent endothelial interval, while amplitudes of SBP oscillations increased in NO-independent and NO-dependent intervals. Hypoxia reduced the [NO]i oscillations amplitude. Heating test during hypoxia increased NO-independent endothelial SBP oscillations and decrease myogenic ones, did not effect on [NO]i oscillations, and shifted [Ca2+]i oscillations peak from 0.005-0.01 Hz to 0.01-0.018 Hz. We observed the [Ca2+]i and [NO]i oscillations synchronization within a cell and between cells for the first time. Heating abolished these synchronizations. Therefore low frequency [Ca2+]i and [NO]i oscillations in endotheliocytes may be considerate as modulator of low frequency endothelial SBP oscillations.

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