EFFECTS OF PROTEASE INHIBITORS ON NEURITE OUTGROWTH AND SURVIVAL OF CULTURED NEURONS
| Autor: | Hiroko Kanai |
|---|---|
| Rok vydání: | 1994 |
| Předmět: | |
| Zdroj: | The KITAKANTO Medical Journal. 44:363-371 |
| ISSN: | 1883-6135 0023-1908 |
| Popis: | Some protease inhibitors have been reported to influence the extent of neurite outgrowth and survivals in various types of cultured neurons. In this paper, I have studied effects of several protease inhibitors on the neurite outgrowth and survival of cultured neurons of the rat.For the trophic effect assay, hippocampal neurons and dorsal root ganglion cells were cultured in chemically defined medium. In order to evaluate the number of surviving hipptcampal neurons semiquantitatively, enzyme immunoassay (EIA) for MAP 2 was carried out using anti-MAP 2 antibody. There was a good correlation between the MAP 2 level obtained by the EIA and the number of neuronal cells counted unedr phase contrast microscopy. Therefore, quantified anti-MAP 2 bindings, primarily reflecting neuronal survival in culture, were used as an index of the neurotrophic activity of protease inhibitors. Neuronal cells were cultured in chemically defined medium containing the following protease inhibitors at various concentrations ; soybean trypsin inhibitor (STI), leupeptin, diisofluorophosphate (DFP), α2-maroglobulin (α2M) and α 1-antichymotrypsin (ACT). Among these protease inhibitors, only ACT and α2M showed trophic effects on cultured hippocampal neurons. Anti MAP2 bindings increased with increasing concentrations of ACT ranging from 0.125μg/ml to 4 μg/ml. The addition of α2M at the optimal dose (100μg/ml) to the culture medium caused a four-fold increase in anti-MAP2 bindings of the cultures of hippocampal neurons. On the other hand, the protease inhibitors tested showed no trophic effects on dorsal root ganglion cells.Under phase-cotrast microscopic observation, only ACT increased both the length and numbers of neurites in cultured neurons. The maximal effect on the increase in neurite length was detected at a concentration of 8 μg/ml.This study has revealed that ACT enhances both survival and neurite outgrowth of hippocampal neurons. It is suggested that ACT, an associated protein of senile plaque amyloid, may act as a trophic factor in the brain of Alzheimer type dementia. |
| Databáze: | OpenAIRE |
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