PMID:14766239

Citation	Li Volti, G, Ientile, R, Abraham, NG, Vanella, A, Cannavò, G, Mazza, F, Currò, M, Raciti, G, Avola, R and Campisi, A (2004) Immunocytochemical localization and expression of heme oxygenase-1 in primary astroglial cell cultures during differentiation: effect of glutamate. Biochem. Biophys. Res. Commun. 315:517-24
Abstract	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 (100microM). 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.
Links	PubMed Online version:10.1016/j.bbrc.2004.01.090
Keywords	Animals; Astrocytes/enzymology; Astrocytes/metabolism; Benzodiazepines/pharmacology; Blotting, Western; Cell Differentiation; Cell Nucleolus/metabolism; Cell Nucleus/metabolism; Cells, Cultured; Cytosol/metabolism; Densitometry; Excitatory Amino Acid Antagonists/pharmacology; Glutamate-Ammonia Ligase/metabolism; Glutamates/metabolism; Glutamic Acid/pharmacology; Heme Oxygenase (Decyclizing)/biosynthesis; Heme Oxygenase-1; Immunoblotting; Immunohistochemistry; Microscopy, Confocal; Microscopy, Fluorescence; Models, Biological; Protein Transport; Rats; Rats, Wistar; Receptors, AMPA/metabolism; Time Factors
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
RAT:HMOX1	part_of	GO:0005730: nucleolus	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
RAT:HMOX1	part_of	GO:0005634: nucleus	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
RAT:HMOX1	part_of	GO:0005829: cytosol	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete

References

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