title: Influence of insulin on glutamine synthetase in the Müller glial cells of retina.
abstract: Glutamine synthetase (GS), a Müller cell specific enzyme in the retina, is the key enzyme involve in glutamate metabolism. The goal of this study was to investigate the expression and regulation of GS by insulin in the cultured rat retinal Müller cells. Immunocytochemical and immunoblotting experiments showed that the cultured Müller cells express GS protein under normal cell culture conditions. Insulin treatments decreased the GS expression both in a time and dose dependent manner. Insulin also decreased the hydrocortisone induced GS expression. Furthermore, we investigated the expression and regulation of two other Müller cell specific enzymes known to be involved in glutamate metabolism, the mitochondrial branched chain aminotransferase (BCATm) and pyruvate carboxylase (PC). Immunoblotting experiments showed that Müller cells expressed both BCATm and PC. Treatments of cells with hydrocortisone or insulin did not influence the BCATm expression level. Hydrocortisone treatment of cells increased the PC expression but this induced expression was suppressed by insulin treatment. Müller cells expressed insulin receptor proteins (IRβ and IRS-1) and insulin activation induced the phosphotyrosine level of insulin receptor proteins. Moreover, hydrocortisone did not influence the expression or activation of these receptor proteins. The data suggests that insulin modulates the GS synthesis and may influence glutamate metabolism in the cultured retinal Müller cells but not by influencing the insulin signaling pathway.
authors.affiliation: Department of Ophthalmology, College of Medicine, King Saud University, Old Airport Road, KAUH, P.O. Box 245, Riyadh 11411, Kingdom of Saudi Arabia. mola@ksu.edu.sa
authors.forename: Mohammad Shamsul
authors.initials: MS
authors.lastname: Ola
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Hosoya
authors.forename: Kathryn F
authors.initials: KF
authors.lastname: LaNoue
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D005974:Glutamate-Ammonia Ligase / Q000096:biosynthesis / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D006854:Hydrocortisone / Q000378:metabolism; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism*; D055504:Insulin Receptor Substrate Proteins / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism; D024101:Mitochondrial Proteins; D027501:Monocarboxylic Acid Transporters; D011766:Pyruvate Carboxylase / Q000378:metabolism*; D051381:Rats; D011972:Receptor, Insulin / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000201:enzymology; D000070590:Solute Carrier Proteins; D000637:Transaminases / Q000378:metabolism
chemical_list: D007328:Insulin; D055504:Insulin Receptor Substrate Proteins; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D027501:Monocarboxylic Acid Transporters; C000629174:Mpc1 protein, rat; D000070590:Solute Carrier Proteins; D018698:Glutamic Acid; D000637:Transaminases; C000629171:Bcat2 protein, rat; C019798:branched-chain-amino-acid transaminase; D011972:Receptor, Insulin; D005974:Glutamate-Ammonia Ligase; D011766:Pyruvate Carboxylase; D006854:Hydrocortisone