eprintid: 10405 rev_number: 69 eprint_status: archive userid: 598 dir: disk0/00/01/04/05 datestamp: 2008-11-05 11:44:25 lastmod: 2015-07-23 09:35:15 status_changed: 2008-11-05 11:44:25 type: article metadata_visibility: show item_issues_count: 0 creators_name: Abramov, A.Y. creators_name: Jacobson, J. creators_name: Wientjes, F. creators_name: Hothersall, J. creators_name: Canevari, L. creators_name: Duchen, M.R. creators_id: AABRA58 creators_id: JJACO94 creators_id: creators_id: JSHOT73 creators_id: LCANE26 creators_id: MRDUC42 title: Expression and modulation of an NADPH oxidase in mammalian astrocytes ispublished: pub subjects: 25700 subjects: 10000 subjects: 3900 divisions: F84 keywords: NADPH oxidase, intracellular calcium, free radicals, oxidative stress, alzheimer's disease, amyloid β peptide note: Published by the Society of Neuroscience This work is licensed under the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. The license allows you to copy, distribute, and transmit the work, as well as adapting it. However, you must attribute the work to the author (but not in any way that suggests that they endorse you or your use of the work), and cannot use the work for commercial purposes without prior permission of the author. If you alter or build upon this work, you can distribute the resulting work only under the same or similar license to this one. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/ or send a letter to Creative Commons, 444 Castro Street, Suite 900, Mountain View, California, 94041, USA. abstract: Amyloid β peptides generate oxidative stress in hippocampal astrocytes through a mechanism sensitive to inhibitors of the NADPH oxidase [diphenylene iodonium (DPI) and apocynin]. Seeking evidence for the expression and function of the enzyme in primary hippocampal astrocytes, we confirmed the expression of the subunits of the phagocyte NADPH oxidase by Western blot analysis and by immunofluorescence and coexpression with the astrocyte-specific marker glial fibrillary acidic protein both in cultures and in vivo. Functional assays using lucigenin luminescence, dihydroethidine, or dicarboxyfluorescein fluorescence to measure the production of reactive oxygen species (ROS) demonstrated DPI and apocynin-sensitive ROS generation in response to the phorbol ester PMA and to raised [Ca2+]c after application of ionomycin or P2u receptor activation. Stimulation by PMA but not Ca2+ was inhibited by the protein kinase C (PKC) inhibitors staurosporine and hispidin. Responses were absent in transgenic mice lacking gp91phox. Expression of gp91phox and p67phox was increased in reactive astrocytes, which showed increased rates of both resting and stimulated ROS generation. NADPH oxidase activity was modulated by intracellular pH, suppressed by intracellular alkalinization, and enhanced by acidification. The protonophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone suppressed basal ROS generation but markedly increased PMA-stimulated ROS generation. This was independent of mitochondrial ROS production, because it was unaffected by mitochondrial depolarization with rotenone and oligomycin. Thus, the NADPH oxidase is expressed in astrocytes and is functional, activated by PKC and intracellular calcium, modulated by pHi, and upregulated by astrocyte activation. The astrocytic NADPH oxidase is likely to play important roles in CNS physiology and pathology. date: 2005-10-05 official_url: http://dx.doi.org/10.1523/JNEUROSCI.1632-05.2005 vfaculties: VFBRS rae2008: 14 oa_status: green language: eng primo: open primo_central: open_green doi: 10.1523/JNEUROSCI.1632-05.2005 lyricists_name: Abramov, A lyricists_name: Transfer, MM lyricists_id: AABRA58 lyricists_id: JJACO94 full_text_status: public publication: Journal of Neuroscience volume: 25 number: 40 pagerange: 9176-9184 refereed: TRUE issn: 0270-6474 citation: Abramov, A.Y.; Jacobson, J.; Wientjes, F.; Hothersall, J.; Canevari, L.; Duchen, M.R.; (2005) Expression and modulation of an NADPH oxidase in mammalian astrocytes. Journal of Neuroscience , 25 (40) pp. 9176-9184. 10.1523/JNEUROSCI.1632-05.2005 <https://doi.org/10.1523/JNEUROSCI.1632-05.2005>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10405/1/10405.pdf