The presence of an extractable substance in the CSF of humans with cerebral vasospasm after subarachnoid haemorrhage that correlates with phosphatase inhibition.
The cellular events leading to cerebral vasospasm after subarachnoid haemorrhage are poorly understood, although an increase in smooth muscle myosin light chain phosphorylation has been observed. This study set out to determine if phosphatase inhibition may be involved in the pathological maintenance of tension observed during vasospasm. We found that 1 nM okadaic acid, a type 2A protein phosphatase inhibitor, elicited an increase in rate of O(2) consumption in the porcine carotid artery similar to that by cerebrospinal fluid (CSF) from vasospastic patients (CSF(V), n=5) (control 0.23+/-0.03, CSF(V) 0.84+/-0.16 and okadaic acid 0.85+/-0.02 micromol min(-1) g dwt(-1)). It was also observed that phosphatase inhibition with 1 nM okadaic acid significantly slowed relaxation after a stretch in a similar fashion to CSF(V) haemorrhage. CSF from vasospastic subarachnoid haemorrhage patients, but not from those without vasospasm, contains an extractable substance which modulates myosin light chain phosphorylation in vitro. A phosphatase preparation obtained from the porcine carotid artery dephosphorylated 63+/-2% of the phosphorylated (MLC(20)) substrate in vitro, and non-vasospastic CSF treated enzyme dephosphorylated 60+/-2.6%. Okadaic acid inhibited phosphatase dephosphorylated only 7.5+/-1% of the substrate where CSF(V) treated enzyme dephosphorylated 22+/-2.8% of the substrate. We conclude that inhibition of smooth muscle phosphatase may be involved in the mechanisms associated with cerebral vasospasm after subarachnoid haemorrhage.
Pyne
G J
GJ
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Cadoux-Hudson
T A
TA
Clark
J F
JF
D000818:Animals; D002339:Carotid Arteries / Q000187:drug effects / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000187:drug effects; D018994:Myosin Light Chains / Q000378:metabolism; D019319:Okadaic Acid / Q000494:pharmacology; D010101:Oxygen Consumption / Q000187:drug effects; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D013345:Subarachnoid Hemorrhage / Q000150:complications*; D013552:Swine; D014664:Vasodilation / Q000187:drug effects; D020301:Vasospasm, Intracranial / Q000134:cerebrospinal fluid / Q000209:etiology*
D004791:Enzyme Inhibitors; D018994:Myosin Light Chains; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases