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The Journal of Neuroscience, October 15, 2002, 22(20):8961-8970
Synaptic Localization of Nitric Oxide Synthase and Soluble Guanylyl Cyclase in the Hippocampus
Alain Burette1, Ulrike Zabel2, Richard J. Weinberg1, Harald H. H. W. Schmidt3, and Juli G. Valtschanoff1 1 Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, North Carolina 27599, 2 Department of Pharmacology and Toxicology, University of Würzburg, 97078 Würzburg, Germany, and 3 Rudolf-Buchheim-Institute for Pharmacology, D-35392 Giessen, Germany
Functional evidence suggests that nitric oxide released from CA1 pyramidal cells can act as a retrograde messenger to mediate hippocampal long-term potentiation, but the failure to find neuronal nitric oxide synthase (NOS-I) in the dendritic spines of these cells has cast doubt on this suggestion. We hypothesized that NOS-I may be in spines but in a form inaccessible to antibody when using standard histological fixation procedures. Supporting this hypothesis, we found that after a weak fixation protocol shown previously to enhance staining of synaptic proteins, CA1 pyramidal cells exhibit clear immunoreactivity for NOS-I. Confocal microscopy revealed that numerous dendritic spines in the stratum radiatum contained the NR2 subunit of the NMDA receptor and the adaptor protein postsynaptic density-95, and a subset of these spines also contained NOS-I. Quantitative studies showed that only ~8% of synaptic puncta (identified by synaptophysin staining) were associated with NOS-I, and ~9% contained the
subunit of soluble guanylyl cyclase (sGC), a major target of NO. However, the majority of NOS-I-positive synaptic puncta was associated with sGC and vice versa. Postembedding immunogold electron microscopy showed that NOS-I concentrates just inside the postsynaptic plasma membrane of asymmetric axospinous synapses in the stratum radiatum of CA1, whereas sGC
Key words: NOS; retrograde messenger; long-term potentiation; sGC; PSD-95; NR2
Copyright © 2002 Society for Neuroscience 0270-6474/02/22208961-10$05.00/0
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