Glyoxal inactivates glutamate transporter-1 in cultured rat astrocytes.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive motor paralysis and selective motor neuron death. There is increasing evidence that motor neuron death in ALS is mediated by glutamate toxicity resulting from reduced activity of astrocytic glutamate transporter-1 (GLT-1). Recent morphological studies have shown that Nepsilon-(carboxymethyl)lysine (CML) accumulates in reactive astrocytes of ALS spinal cords. CML is a product of post-translational protein modification by glyoxal, a reactive aldehydic intermediate. In considering these documents, it is important to determine whether GLT-1 protein modification by glyoxal might cause reduced GLT-1 activity. To address this issue, we investigated the effects of glyoxal on GLT-1 properties in cultured rat astrocytes. High performance liquid chromatography showed reduced glutamate uptake activity in the glyoxal-exposed cells. Immunocytochemical analysis displayed CML accumulation in the cytoplasm of astrocytes by glyoxal exposure. Immunoblots of immunoprecipitated GLT-1 disclosed GLT-1 CML adduct formation in the glyoxal-exposed cells. Our results indicate that glyoxal modifies GLT-1 to form CML and simultaneously deprives its glutamate uptake activity. Thus, these toxic effects of glyoxal on astrocytes might be implicated in motor neuron death in ALS.
Kawaguchi
Motoko
M
Department of Pathology, Tokyo Women's Medical University, Tokyo, Japan. m-kawa@research.twmu.ac.jp
Shibata
Noriyuki
N
Horiuchi
Seikoh
S
Kobayashi
Makio
M
D027322:Amino Acid Transport System X-AG / Q000187:drug effects* / Q000378:metabolism; D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism / Q000503:physiopathology; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D018698:Glutamic Acid / Q000187:drug effects / Q000378:metabolism*; D006037:Glyoxal / Q000633:toxicity*; D015151:Immunoblotting; D007150:Immunohistochemistry; D008239:Lysine / Q000031:analogs & derivatives* / Q000378:metabolism; D051381:Rats
D027322:Amino Acid Transport System X-AG; D018698:Glutamic Acid; D006037:Glyoxal; C048496:N(6)-carboxymethyllysine; D008239:Lysine