Immunocytochemical localization and expression of heme oxygenase-1 in primary astroglial cell cultures during differentiation: effect of glutamate
| Autor: | Angelo Vanella, Roberto Avola, Monica Currò, Nader G. Abraham, Riccardo Ientile, Giuseppe Cannavò, Agata Campisi, Francesco Mazza, Giovanni Li Volti, Giuseppina Raciti |
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| Rok vydání: | 2004 |
| Předmět: |
Time Factors
Blotting Western Immunoblotting Biophysics Glutamic Acid AMPA receptor Biology Models Biological Biochemistry Benzodiazepines Cytosol Glutamates Western blot Glutamate-Ammonia Ligase medicine Animals Receptors AMPA Rats Wistar Receptor Molecular Biology Cells Cultured Cellular compartment Cell Nucleus Microscopy Confocal medicine.diagnostic_test Heme Oxygenase-1 Glutamate astroglial cells Glutamate receptor Cell Differentiation Cell Biology Subcellular localization Immunohistochemistry Molecular biology Rats Cell biology Heme oxygenase Protein Transport Microscopy Fluorescence Astrocytes Heme Oxygenase (Decyclizing) Excitatory Amino Acid Antagonists Cell Nucleolus Densitometry |
| Zdroj: | Biochemical and Biophysical Research Communications. 315:517-524 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2004.01.090 |
| Popis: | Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation releasing iron, carbon monoxide (CO), and biliverdin. We investigated subcellular localization of HO-1 using confocal laser scanning microscopy (CLSM) and the expression by Western blot in primary astroglial cells during differentiation and after exposure to glutamate (100 μM). CLSM analysis of immunostained HO-1 in cultured astroglial cells during differentiation showed an increase of fluorescence between 7 and 14 days and a decrease between 14 and 21, although HO-1 peaked at 14 days it remained at high levels. The distribution of HO-1 protein undergoes modification in the various cellular compartments. Furthermore, localization of the protein in untreated astrocytes at 7 days appeared prevalently localized in the cytosol and in the perinuclear region. In contrast, at 14 and 21 days, fluorescence detection suggests that HO-1 was present also in the nucleus, and in the nucleoli. Fluorescence intensity significantly increased in glutamate-treated astrocytes during all development stages and the protein appeared in the cytosol, in the nucleus and in the nucleoli. The involvement of AMPA/Ka receptors was studied in glutamate-treated astroglial cells at 14 days by the preincubation of the cells with GYKI 52466, a specific receptor inhibitor, of AMPA/Ka receptor demonstrating the involvement of these receptors. Western blot analysis of HO-1 confirmed the CLSM results. Our results demonstrate that changes in HO-1 protein expression and localization in primary cultured astroglial cells may be part of the underlying mechanisms involved in brain development as well as in neurodegenerative diseases. |
| Databáze: | OpenAIRE |
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