Thrombin, its receptor and protease nexin I, its potent serpin, in the nervous system

Abstract

The multifunctional serine protease, thrombin, the principal component of the blood coagulation cascade, is also active in nervous system growth and maintenance. In neural tissue culture, it prevents neurite outgrowth and modulates morphologic changes in both neurons and astrocytes. In recent studies, we found that it mediates polyneuronal synapse elimination, both in vivo and in vitro. Of relevance to neurologic disease, as well as to development, evidence also implicates thrombin in apoptosis of these cells. As with other serine proteases, thrombin is in "balance" with one or more endogenous protein inhibitors, members of the serpin superfamily of proteins. The most potent vertebrate inhibitor for thrombin is protease nexin I (PNI), which regulates thrombin's effect by forming post-translational, covalent complexes with the protease. We review some of the nervous system effects of the thrombin:PNI balance, and also present results of a recent study of this balance after peripheral nerve injury. We measured thrombin and prothrombin activity in extracts from adult mouse sciatic nerve using a specific chromogenic assay. We also performed reverse transcription polymerase chain reaction of RNA from nerve crush samples. We found a burst of activity at 3 days following injury distal to the crush site that was inhibited by thrombin specific inhibitors. It is possible that a significant fraction of the increased prothrombin in injured nerve was synthesized locally. Active PNI levels increased in these crush samples 6 to 9 days after the thrombin induction. These data suggest that nerve injury first induces the synthesis of prothrombin, which is subsequently converted to active thrombin. Nerve crush-induced thrombin is followed by the generation of functionally active PNI and may be directly responsible for its induction. These results suggest that the balance between serine proteases and their serpins is dysregulated during nerve injury and support a role for its reestablishment in nerve damage repair.