The serine protease subtilisin suppresses epileptiform activity in rat hippocampal slices and neocortex in vivo.

Serine proteases of the S8A family and those belonging to the subtilase group generate a long-lasting inhibition of hippocampal evoked potentials, which shows little recovery and resembles long-term depression. The present work investigates the effects of subtilisin A on epileptiform activity induced in hippocampal slices. Interictal bursts were generated by perfusion with 4-aminopyridine in magnesium-free medium, whereas ictal bursts were produced by the addition of baclofen. Subtilisin A superfused for 10 min at concentrations of 50 nM and above reduced the duration of ictal bursts, whereas higher concentrations reduced the frequency of interictal activity with little or no recovery, indicating similarity with the long-term depression reported previously. The anti-epileptiform activity was not prevented by inhibitors of phosphatases or several kinases, but the inhibition of ictal activity was selectively reduced by the tyrosine kinase inhibitor genistein. The rho-activated coiled-coil kinase (ROCK) inhibitor Y-27632 had no effect on the suppression of ictal or interictal bursts. Subtilisin applied at nanomolar concentrations to the surface of the cerebral cortex in vivo also suppressed epileptiform spikes induced by bicuculline. It is concluded that serine proteases of the subtilase group are highly potent inhibitors of epileptiform activity, especially ictal bursts, and that tyrosine kinases may be involved in that inhibition. The mechanism of inhibition is different from the long-lasting depression of evoked potentials, which is partly mediated via ROCK.