TY - JOUR
T1 - Role of NADPH oxidase in the apoptotic death of cultured cerebellar granule neurons
AU - Coyoy, Angélica
AU - Valencia, Antonio
AU - Guemez-Gamboa, Alicia
AU - Morán, Julio
N1 - Funding Information:
The authors are grateful to Guadalupe Dominguez for excellent technical assistance. This work was supported by DGAPA-UNAM (IN223107) and CONACYT (47158Q). Angelica Coyoy was supported by a CONACYT fellowship.
PY - 2008/10/15
Y1 - 2008/10/15
N2 - Cerebellar granule neurons (CGN) cultured in a medium containing 25 mM KCl and treated with staurosporine (ST) or transferred to a medium with 5 mM KCl (K5) die apoptotically. CGN death is mediated by an increase in reactive oxygen species (ROS) production. When CGN are treated with antioxidants all apoptotic parameters and cell death are markedly diminished, showing a central role for ROS in this process. Recently, it has been suggested that a possible ROS source involved in cell death is a NADPH oxidase. In that regard, we found expression in CGN of the components of NADPH proteins, p40phox, p47phox and p67phox, and p22phox, as well as three homologues of the catalytic subunit of this complex, NOX1, 2, and 4. The inhibition of NADPH oxidase with diphenylene iodonium or 4-(2-aminoethyl)benzenesulfonyl fluoride significantly reduced ROS production, NADPH oxidase activity, all the apoptotic events, and cell death induced by both K5 and ST. We conclude that ROS could be an early signal of apoptotic neuronal death and that NADPH oxidase, including NOX1, 2, and/or 4, could have a central role in apoptotic death induced by different conditions in these neurons.
AB - Cerebellar granule neurons (CGN) cultured in a medium containing 25 mM KCl and treated with staurosporine (ST) or transferred to a medium with 5 mM KCl (K5) die apoptotically. CGN death is mediated by an increase in reactive oxygen species (ROS) production. When CGN are treated with antioxidants all apoptotic parameters and cell death are markedly diminished, showing a central role for ROS in this process. Recently, it has been suggested that a possible ROS source involved in cell death is a NADPH oxidase. In that regard, we found expression in CGN of the components of NADPH proteins, p40phox, p47phox and p67phox, and p22phox, as well as three homologues of the catalytic subunit of this complex, NOX1, 2, and 4. The inhibition of NADPH oxidase with diphenylene iodonium or 4-(2-aminoethyl)benzenesulfonyl fluoride significantly reduced ROS production, NADPH oxidase activity, all the apoptotic events, and cell death induced by both K5 and ST. We conclude that ROS could be an early signal of apoptotic neuronal death and that NADPH oxidase, including NOX1, 2, and/or 4, could have a central role in apoptotic death induced by different conditions in these neurons.
KW - Apoptosis
KW - Cerebellar granule neurons
KW - Free radicals
KW - Low potassium
KW - NADPH oxidase
KW - NOX
KW - Rat
KW - Reactive oxygen species
KW - Staurosporine
KW - Superoxide anion
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U2 - 10.1016/j.freeradbiomed.2008.06.027
DO - 10.1016/j.freeradbiomed.2008.06.027
M3 - Article
C2 - 18675340
AN - SCOPUS:53049091077
VL - 45
SP - 1056
EP - 1064
JO - Free Radical Biology and Medicine
JF - Free Radical Biology and Medicine
SN - 0891-5849
IS - 8
ER -