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M. Imasawa, K. Kashiwagi, Y. Iizuka, M. Tanaka, S. Tsukahara, H. Iijima; Different Expression Role Among Glutamate Transporters in Rat Retinal Glial Cells Under Various Culture Conditions . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4021.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Using retinal glial cells from cultured rat retinas, the changes in the expression of glutamate transporters (GLTs) under such conditions as varying degrees of confluence of the retinal glial cells, hypoxia, glutamate loading and potassium loading, were assessed. Methods: After retinal glial cells were isolated from three–day–old Sprague–Dawley rats, GLAST, GLT–1 and EAAC1 mRNA expression was confirmed by reverse transcription–polymerase chain reaction (RT–PCR) and their protein expression was confirmed by Western blot analysis and immunostaining. Changes in the expression of GLT at 6 hours after passage and at confluence, during culture in 5% oxygen, during glutamate loading of the medium and during potassium loading of the medium were assessed using real–time PCR. Results: The presence of three GLTs, namely, GLAST, GLT–1 and EAAC1, in cultured retinal glial cells was confirmed by Western blot analysis, immunostaining and RT–PCR. Although the expression of GLAST mRNA was increased during glutamate and potassium loading, no changes in expression were observed as a result of hypoxia and varying degrees of confluence. The expression of GLT–1 mRNA was increased during hypoxia and confluence, but there were no changes in GLT–1 mRNA expression during glutamate and potassium loading. EAAC–1 mRNA was not changes only in glutamate loading. Conclusions: The expression of EAAC1 in cultured retinal glial cells was confirmed. The expression of GLTs varied according to the environment and the type of loading, suggesting that the involvement of GLTs in retinal physiology and pathology varies depending on the subtype.
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