Synaptic localization of a functional NADPH oxidase in the mouse hippocampus.
Superoxide has been shown to be critical for hippocampal long-term potentiation (LTP) and hippocampus-dependent memory function. A possible source for the generation of superoxide during these processes is NADPH oxidase. The active oxidase consists of two membrane proteins, gp91phox and p22phox, and four cytosolic proteins, p40phox, p47phox, p67phox, and Rac. Upon stimulation, the cytosolic proteins translocate to the membrane to form a complex with the membrane components, which results in production of superoxide. Here, we determined the presence, localization, and functionality of a NADPH oxidase in mouse hippocampus by examining the NADPH oxidase proteins as well as the production of superoxide. All of the NADPH oxidase proteins were present in hippocampal homogenates and enriched in synaptoneurosome preparations. Immunocytochemical analysis of cultured hippocampal neurons indicated that all NADPH oxidase proteins were localized in neuronal cell bodies as well as dendrites. Furthermore, double labeling analysis using antibodies to p67phox and the presynaptic marker synaptophysin suggest a close association of the NADPH oxidase subunits with synaptic sites. Finally, stimulation of hippocampal slices with phorbol esters triggered translocation of the cytoplasmic NADPH oxidase proteins to the membrane and an increase in superoxide production that was blocked by inhibitors of NADPH oxidase. Taken together, our data suggest that NADPH oxidase is present in mouse hippocampus and might be the source of superoxide production required for LTP and memory function.
Tejada-Simon
Maria V
MV
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030, USA.
Serrano
Faridis
F
Villasana
Laura E
LE
Kanterewicz
Beatriz I
BI
Wu
Gang-Yi
GY
Quinn
Mark T
MT
Klann
Eric
E
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D003600:Cytosol / Q000201:enzymology; D006624:Hippocampus / Q000378:metabolism*; D007150:Immunohistochemistry; D017774:Long-Term Potentiation / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019255:NADPH Oxidases / Q000378:metabolism*; D009924:Organ Culture Techniques; D010703:Phorbol Esters / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D013481:Superoxides / Q000378:metabolism; D013569:Synapses / Q000201:enzymology*
D010703:Phorbol Esters; D013481:Superoxides; D019255:NADPH Oxidases