NADPH oxidase subunit, gp91phox homologue, preferentially expressed in human colon epithelial cells

Kikuchi, Hideaki; Hikage, Mituru; Matsumoto, Hitoshi; Fukumoto, Manabu

doi:10.1016/s0378-1119(00)00258-4

Cited by 135 publications

(108 citation statements)

References 15 publications

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“…Intracellular regions of the Nox family are composed of highly conserved domains; therefore, specific Ab for Nox1 had not been available. We have developed polyclonal Abs against human Nox1 useful for immunoblot and immunohistochemical analyses, and found that Nox1 protein was constitutively expressed in surface mucous cells of the guinea pig colon, supporting an in situ hybridization study showing that the Nox1 transcript was expressed in epithelial cells of human colonic mucosa (27). A small amount of Nox1 mRNA is detectable in primary cultures of guinea pig gastric mucosal cells even in LPS-free conditions (12,14), while Nox1 protein was absent in normal gastric and small intestinal mucosal tissues of both guinea pigs and humans (data not shown).…”

Section: Discussionsupporting

confidence: 55%

Role of Nicotinamide Adenine Dinucleotide Phosphate Oxidase 1 in Oxidative Burst Response to Toll-Like Receptor 5 Signaling in Large Intestinal Epithelial Cells

Kawahara

Kuwano

Teshima-Kondo

et al. 2004

The Journal of Immunology

127

149

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The NADPH oxidase 1 (Nox1) is a gp91phox homologue preferentially expressed in the colon. We have established primary cultures of guinea pig large intestinal epithelial cells giving 90% purity of surface mucous cells. These cells spontaneously released superoxide anion (O2−) of 160 nmol/mg protein/h and expressed the Nox1, p22phox, p67phox, and Rac1 mRNAs, but not the gp91phox, Nox4, p47phox, p40phox, and Rac2 mRNAs. They also expressed novel homologues of p47phox and p67phox (p41nox and p51nox, respectively). Human colon cancer cell lines (T84 and Caco2 cells) expressed the Nox1, p22phox, p51nox, and Rac1 mRNAs, but not the other NADPH component mRNAs, and secreted only small amounts of O2− (<2 nmol/mg protein/h). Cotransfection of p41nox and p51nox cDNAs in T84 cells enhanced PMA-stimulated O2− release 5-fold. Treatment of the transfected T84 cells with recombinant flagellin (rFliC) from Salmonella enteritidis further augmented the O2− release in association with the induction of Nox1 protein. The enhanced O2− production by cotransfection of p41nox and p51nox vectors further augmented the rFliC-stimulated IL-8 release from T84 cells. T84 cells expressed the Toll-like receptor 5, and rFliC rapidly phosphorylated TGF-β-activated kinase 1 and TGF-β-activated kinase 1-binding protein 1. A potent inhibitor for NF-κB (pyrrolidine dithiocarbamate) significantly blocked the rFliC-primed increase in O2− production and induction of Nox1 protein. These results suggest that p41nox and p51nox are involved in the Nox1 activation in surface mucous cells of the colon, and besides that, epithelial cells discern pathogenicities among bacteria to appropriately operate Nox1 for the host defense.

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Section: Discussionsupporting

confidence: 55%

Role of Nicotinamide Adenine Dinucleotide Phosphate Oxidase 1 in Oxidative Burst Response to Toll-Like Receptor 5 Signaling in Large Intestinal Epithelial Cells

Kawahara

Kuwano

Teshima-Kondo

et al. 2004

The Journal of Immunology

127

149

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“…Cloning of Mouse and Rat NOX3 cDNA from Inner Ear-NOX3 mRNA has been shown to be expressed in human embryonic kidney, but expression levels were very low (23,24); hence, the physiological relevance was questionable. We reasoned that the physiologically relevant localization of NOX3 might have been missed because previous studies had restricted their analysis to commercially available human RNA sources.…”

Section: Resultsmentioning

confidence: 99%

“…colon epithelium for NOX1 (16,17), kidney cortex for NOX4 (18,19), lymphoid organs and testis for NOX5 (20), and the thyroid gland for DUOX1 and DUOX2 (21, 22)). The tissue distribution of NOX3 was systematically investigated in two studies (23,24) in which the authors found only very low levels of NOX3 mRNA expression in embryonic kidney.Our knowledge of the activation mechanisms of the members of NOX/DUOX family varies considerably among individual enzymes. NOX1 and gp91 phox /NOX2 are subunit-dependent en-…”

mentioning

confidence: 99%

NOX3, a Superoxide-generating NADPH Oxidase of the Inner Ear

Bánfi

Malgrange

Knisz³

et al. 2004

Journal of Biological Chemistry

402

349

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Reactive oxygen species (ROS) play a major role in drug-, noise-, and age-dependent hearing loss, but the source of ROS in the inner ear remains largely unknown. Herein, we demonstrate that NADPH oxidase (NOX) 3, a member of the NOX/dual domain oxidase family of NADPH oxidases, is highly expressed in specific portions of the inner ear. As assessed by real-time PCR, NOX3 mRNA expression in the inner ear is at least 50-fold higher than in any other tissues where its expression has been observed (e.g. fetal kidney, brain, skull). Microdissection and in situ hybridization studies demonstrated that NOX3 is localized to the vestibular and cochlear sensory epithelia and to the spiral ganglions. Transfection of human embryonic kidney 293 cells with NOX3 revealed that it generates low levels of ROS on its own but produces high levels of ROS upon co-expression with cytoplasmic NOX subunits. NOX3-dependent superoxide production required a stimulus in the absence of subunits and upon co-expression with phagocyte NADPH oxidase subunits p47 phox and p67 phox , but it was stimulus-independent upon co-expression with colon NADPH oxidase subunits NOX organizer 1 and NOX activator 1. Pre-incubation of NOX3-transfected human embryonic kidney 293 cells with the ototoxic drug cisplatin markedly enhanced superoxide production, in both the presence and the absence of subunits. Our data suggest that NOX3 is a relevant source of ROS generation in the cochlear and vestibular systems and that NOX3-dependent ROS generation might contribute to hearing loss and balance problems in response to ototoxic drugs.The inner ear is a highly complex structure involved in hearing and balancing. The conversion of sound into electrical signals occurs within the cochlea, in the organ of Corti, and the electrical signals are conducted by the axons of spiral ganglion neurons to the brain. The linear movement of the head is sensed by the otolith organs (utricle and saccule) and the rotation movements by the ampulla of the semicircular canals. The signals generated in the vestibular system are transmitted by the vestibular ganglion neurons to the central nervous system.Hearing impairment caused by loss of cochlear function occurs frequently, if not invariably, over a lifetime. Noise and ototoxic chemicals may lead to a precocious, rapid hearing loss, whereas aging leads to a more insidious, chronic loss of hearing. Research over the last decades has identified reactive oxygen species (ROS) 1 as the major factor mediating hearing loss (1). ROS is generated within the cochlea after exposure to ototoxic drugs (e.g. cisplatin (2, 3), aminoglycoside antibiotics (3)) or to noise (4). Signs of oxidative stress, such as DNA damage and lipid peroxidation, have been documented in vivo in response to those challenges (5, 6), as well as in cochlear aging (7). The vestibular system is also damaged by ototoxic drugs (8, 9) in a process that includes excessive ROS production (10, 11).Although the role of oxidative stress in inner ear damage is well established, its source ...

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“…Nox-3 présente 58 % d'homologie avec Nox-2 et comprend 568 aa (Figure 3) [14]. Les ARNm de Nox-3 n'ont pas été détectés dans les tissus adultes.…”

Section: Nox-3unclassified

“…Les ARNm de Nox-3 n'ont pas été détectés dans les tissus adultes. Cependant, des expériences de RT-PCR ont permis de détecter les ARNm de Nox-3 dans différents tissus foetaux tels que le rein, le foie, le poumon et la rate [8,14]. Chez l'adulte, son expression est exclusivement réduite à l'oreille interne et joue un rôle majeur dans la formation des otoconies, dépôts de carbonate de calcium qui reposent sur les cellules sensorielles ciliées de la macule de l'oreille interne.…”

Section: Nox-3unclassified