ABSTRACT
The calpains-calpastatin system (calcium-activated neutral proteases and endogenous inhibitor) seems to be, in the skeletal muscle, a fine enzymatic system of myofibrillar turnover regulation, in normal as well as pathological conditions (for ex., dystrophic muscle). The purpose of the research is to establish in qualitative and quantitative terms whether the level of calpastatin can evidence differences between a muscle in normal activity conditions and one having dysfunctional alterations experimentally induced. So the masseter muscle of rabbit in normal conditions and with experimentally modified occlusal plane has been used. Our results confirm the presence of the 68 KDa calpastatin in the masseter muscle. The presence of the inhibitor in the same subcellular structures in which the calpains have been detected (myofibrillars, sarcolemma, endomysial connective) has been confirmed. Finally, variations in calpastatin amount in the muscle of animals experimentally treated with respect to the controls have been found. Thus, calpastatin seems to act as a marker of muscle dysfunctions connected to occlusal plane alteration and to loss of vertical dimention.
Subject(s)
Calcium-Binding Proteins/pharmacokinetics , Calpain/pharmacokinetics , Cysteine Proteinase Inhibitors/pharmacokinetics , Masseter Muscle/chemistry , Animals , Blotting, Western , Calcium-Binding Proteins/immunology , Calpain/immunology , Cysteine Proteinase Inhibitors/immunology , Enzyme-Linked Immunosorbent Assay , Immunochemistry , Immunohistochemistry , Malocclusion/physiopathology , Masseter Muscle/immunology , Rabbits , Tooth Abrasion/physiopathology , Vertical DimensionABSTRACT
The purpose of this experiment was to assess the roles of free, intracellular calcium and calcium-dependent neutral protease (calpain II, EC.34.22.17) in selenite nuclear cataract. Free calcium ion concentrations within lens nuclear fibers during selenite cataractogenesis increased to 3 microM on day 2 post-injection (clear lens) and to 108 microM at day 4 (nuclear cataract). Calpain II is known to be activated in vitro by calcium levels above 50 microM. Calpain II activity was present in the lens nucleus at time periods preceding formation of selenite cataract. These data suggested that after selenite injection, calpain II was activated by elevated free calcium in the nucleus, and that calpain II-induced proteolysis of nuclear proteins was an important mechanism in selenite cataract. Calpain II levels were also observed to decrease in the nucleus during selenite cataractogenesis, probably due to autolysis. This was supported by the finding that incubation of purified lens calpain II with 100 microM calcium caused partial inactivation of the protease.
