[Abstract] Objective: To investigate the effect of glial fibrillary acidic protein and glutamine synthetase in retina Müller cell in vitro on transporting Glutamate in the condition of high concentration of glutamate. Methods: 1 The primary retinal cells were purified by the constant-temperature shaking method. Müller cell was identified by immunocytochemistry staining of GFAP and GS. Afterwards we observed the general appearance by Hematoxylin and Eosin staining (H-E staining) 2 50mM glutamate was added in the early passaged medium 1 day and 4 days before determing the activity of Müller cells by MTT chromatometry. Then we observed the general appearance by H-E staining again. We performed immunocytochemistry staining of GFAP and GS on Müller cell again , and compared the difference of the expression of them by computerized micro-imaging analysis system. Results: The activity of Müller cell was enhanced, the expression of GFAP was improved, but there was no significant difference in morphological changes and the expression of GS. Conclusions: The constant temperature shaking method provides a rapid and reliable method for purification of retinal Müller cells. GFAP、 GS were expressed in retinal Müller cells in normal physiological condition in vitro. GFAP takes part in transporting of glutamate and will increase in the early period when the cell is stressed.GS ,the key enzyme in transformating glutamate to glutamine,also takes part in transporting of glutamate, and will not increase in the early period when the cell is stressed. In some pathological changes the concentration of extracellular will increase. The increase in transporting but lag in metabolism is motivation of cytotoxic effect of glutamate, which causes the cell damage.
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