Myosin Light Chain Phosphorylation and Contractile Proteins in a Canine Two-Hemorrhage Model of Subarachnoid Hemorrhage

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Abstract

Background and Purpose

Subarachnoid hemorrhage (SAH) impairs both contraction and relaxation response in cerebral arteries. We tested the hypothesis that cerebral vasospasm might be ATP-independent contraction, such as latch state, and protein synthesis might be substantially downregulated due to ATP consumption after long-lasting contraction.

Methods

Chronic cerebral vasospasm was induced in the canine 2-hemorrhage model of SAH. The normal and spastic basilar arteries were stabilized in Krebs-Henseleit solution, and contraction was induced by 30 [micro sign]mol/L prostaglandin F2 alpha (PGF2 alpha) in vitro and in vivo. Before and at 15 minutes and 1 hour after the treatment with PGF2 alpha, the levels of phosphorylated 20-kDa myosin light chain (MLC20) were measured. The time course of expression of contraction proteins actin and MLC20, and contraction-inhibiting proteins h-caldesmon and calponin was determined by immunoblotting techniques.

Results

A significant vasospasm occurred in the basilar artery during days 4 to 21, most prominently on days 7 and 14. There were no significant differences in the baseline levels of phosphorylated MLC20 between normal and spastic basilar arteries. The increase in MLC20 phosphorylation by PGF2 alpha was significantly attenuated in the spastic basilar artery in vitro and in vivo (P<0.05). The immunoreactivity for actin, h-caldesmon, and calponin in the spastic basilar arteries was progressively decreased until day 14 and returned to the normal level on day 21. In contrast, protein levels of MLC20 did not significantly change during days 0 to 21.

Conclusions

Chronic cerebral vasospasm closely resembles the latch state, and temporary deficiencies of contractile proteins may result from increased destruction and inhibition of protein synthesis. (Stroke. 1998;29:2149-2154.)

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