Glutamate-induced increases in transglutaminase activity in primary cultures of astroglial cells
| Autor: | V. Macaione, Giuseppina Raciti, Monica Currò, Daniela Caccamo, Salvatore Macaione, Angelo Vanella, Riccardo Ientile, Agatina Campisi, Giuseppe Cannavò |
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| Rok vydání: | 2003 |
| Předmět: |
Time Factors
Tissue transglutaminase Cellular differentiation Blotting Western Excitotoxicity Glutamic Acid AMPA receptor Biology medicine.disease_cause chemistry.chemical_compound Benzodiazepines Glutamate-Ammonia Ligase Cadaverine medicine Animals Drug Interactions Excitatory Amino Acid Agents Rats Wistar Molecular Biology Cells Cultured Microscopy Confocal Transglutaminases Dose-Response Relationship Drug General Neuroscience Acridine orange Glutamate receptor Molecular biology Immunohistochemistry Rats medicine.anatomical_structure chemistry Animals Newborn Anti-Anxiety Agents Astrocytes biology.protein Neuroglia Neurology (clinical) Excitatory Amino Acid Antagonists Developmental Biology Astrocyte |
| Zdroj: | Brain research. 978(1-2) |
| ISSN: | 0006-8993 |
| Popis: | Glutamate exposure of astroglial cells caused ligand-gated channel receptor activation, associated with excitotoxic cell response. We investigated the effects of 24 h glutamate exposure on transglutaminase in astrocytes primary cultures at 7, 14, and 21 days in vitro (DIV). Increases in enzyme activity were observed as a function of cell differentiation stage in glutamate-treated cultures. These effects were significantly reduced when GYKI 52466, an AMPA/KA receptors inhibitor, was added to the culture medium prior to incubation with glutamate. Microscopy observation on transglutaminase-mediated, fluorescent dansylcadaverine incorporation in living cells was consistent with these results. Western blotting analysis with monoclonal antibody showed that glutamate also up-regulated tissue transglutaminase expression, which reached the highest values in 14 DIV cultures. Confocal laser scanning microscopy analysis of immunostained astroglial cells showed a mainly cytoplasmic localisation of the enzyme both in control and treated cultures; nevertheless, counterstaining with the nuclear dye acridine orange demonstrated the presence of tissue transglutaminase also into the nucleus of glutamate-exposed and 21 DIV cells. The increases in enzyme expression and localisation in the nucleus of glutamate-treated astroglial cells may be part of biochemical alterations induced by excitotoxic stimulus. |
| Databáze: | OpenAIRE |
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