Glutamine synthetase becomes nitrated and its activity is reduced during repetitive seizure activity in the pentylentetrazole model of epilepsy

• Published in: Epilepsia (Copenhagen) Vol. 49; no. 10; pp. 1733 - 1748
• Main Authors: Bidmon, Hans‐J., Görg, Boris, Palomero‐Gallagher, Nicola, Schleicher, Axel, Häussinger, Dieter, Speckmann, Erwin J., Zilles, Karl
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.10.2008; Wiley-Blackwell
• Subjects: Animals; Anticonvulsants. Antiepileptics. Antiparkinson agents; Astrocytes; Astrocytes - drug effects; Astrocytes - metabolism; Biological and medical sciences; Brain - metabolism; Brain - pathology; Disease Models, Animal; Epilepsy - chemically induced; Epilepsy - metabolism; Epilepsy - pathology; Epilepsy - physiopathology; Gene Expression Regulation - drug effects; Glial Fibrillary Acidic Protein - metabolism; Glutamate metabolism; Glutamate-Ammonia Ligase - metabolism; Headache. Facial pains. Syncopes. Epilepsia. Intracranial hypertension. Brain oedema. Cerebral palsy; HSP27 Heat-Shock Proteins - metabolism; HSP‐27; Immunoprecipitation - methods; Male; Medical sciences; Methionine‐sulfoximine; Models, Biological; Nervous system (semeiology, syndromes); Neurology; Neuropharmacology; Nitrotyrosine; Oxidative stress; Pentylenetetrazole; Pharmacology. Drug treatments; Rats; Rats, Wistar; Oxidative stress; Thiol; Nervous system diseases; Astrocytes; Glutamate metabolism; Neuroglia; Epilepsy; Methionine; Astrocyte; Glutamate; Metabolism; Cerebral disorder; Sulfur containing aminoacid; Convulsion; Central nervous system disease; Excitatory aminoacid; Neurotransmitter; Methionine-sulfoximine; Models; Nitrotyrosine; Neurological disorder; HSP-27; Glutamine
• ISSN: 0013-9580; 1528-1167
• DOI: 10.1111/j.1528-1167.2008.01642.x

Summary Purpose:  The astrocyte‐specific glutamine synthetase (GS) plays a key role in glutamate recycling and Gamma‐aminobutyric acid (GABA) metabolism. Changes in the expression or activity of GS have been proposed to contribute to epileptogenesis. The mechanisms or how and where GS may contribute to epilepsy is still a matter of discussion. Here we asked the question whether brain regions, which show an astrocytic stress response respond with alterations of GS. Methods:  Biochemical and histological alterations of GS, HSP‐27, and GFAP were studied after pentylenetetrazole‐induced repetitive epileptic seizures (PIRS) in rats using a topographical quantification of the GS‐immunoreactivity (GSIR) in relation to the focal heat shock response (HSR). Saline‐treated rats served as controls and rats treated by the GS‐inhibitor, L‐methionine‐sulfoximine (MSO) served as a positive control. Results:  No changes in the amount of GSIR and GS‐protein occurred during PIRS. A significant reduction of GSIR was observed by histochemistry (in situ) and in native (nonheated) protein extracts of MSO‐treated rats. In rats affected by PIRS, GS‐activity showed a significant, region‐specific reduction in association with a nitration of the enzyme. Discussion:  These results show that neither PIRS nor GS‐inhibition reduced the amount of GS protein, but that MSO interferes with antibody binding to native GS. PIRS resulted in a focal increase of astrocytic stress response, whereas MSO caused a widespread, homogeneous astrocytic HSR independent from quantitative changes of GS content. In rats with PIRS the regions showing a strong glial HSR, respond with reduced GS‐activity and GS‐nitration, which all together are clear indicators of a nitrosative stress response.