Evidence for decrease in myofilament responsiveness to Ca2+ during hypoxia in spontaneously active vascular smooth muscle in rats

The cellular mechanisms underlying smooth muscle hypoxic relaxation were examined in experiments on isolated chemically skinned muscle strips of rat portal vein. Low PO2 (3.9 kPa) shifted the pCa-tension relation to the right by 0.29 +/- 0.01 pCa units as compared to control curves (PO2, 18.8 kPa). Thus the Ca2+ sensitivity of the filaments had decreased. Addition of cyclic AMP (30 mumol l-1) with theophylline (5 mmol l-1) to the buffer solution produced a similar shift to low PO2. Low PO2 also decreased the maximal force to 40-50% of control. Inclusion of sodium fluoride (10 mmol l-1) and aluminium chloride (10 mumol l-1) in a bath solution caused partial (25-30%) relaxation of skinned smooth muscle preconstricted with 10 mumol l-1 Ca2+. The rate and amplitude of smooth muscle relaxation at low PO2 were significantly decreased by tolbutamide (5 mmol l-1), which is known to inhibit cyclic AMP-dependent protein kinases. We suggest that PO2 changes can alter myofilament responsiveness to Ca2+ and this effect may be related to cyclic AMP-dependent phosphorylation of myosin light chain kinase, its inactivation and subsequent uncoupling between Ca2+ and contractile machinery in smooth muscle.