Abstract：
:Nitric oxide (NO) is a gas messenger with diverse physiological roles in the nervous system, from modulation of synaptic plasticity and neurogenesis to the mediation of neuronal death. NO production in the brain is catalyzed by three isoforms of NO synthase (NOS) including neuronal NOS (nNOS), inducible NOS and endothelial NOS. In this report, we demonstrate a method for in vitro and in vivo silencing of nNOS using RNAi strategies. Because of their efficiency in infecting postmitotic cells like neurons, lentiviral vectors were used as nNOS shRNA carriers. Of the siRNA sequences screened, one corresponding to exon 10 of the rat nNOS specifically and efficiently inhibited nNOS expression at the mRNA and protein level. In vitro experiments using rat cortical neurons showed the general efficacy of shRNA vectors in silencing constitutively expressed nNOS. To demonstrate the anatomical specificity of nNOS silencing in vivo, vectors were used to selectively knock-down the endogenous nNOS expression in cortical GABAergic interneurons of rat piriform cortex. Our findings show that the method reported here can achieve stable and highly effective nNOS suppression in an anatomically defined brain region. The ability of our nNOS silencing vectors to effectively and precisely silence nNOS expression shows their value as research tools for further studies of the role of nNOS in specific brain circuits. Furthermore, our findings raise the possibility for future considerations of lentiviral strategies as therapies for diseases of the nervous system involving NO neurotoxic cascades.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Mahairaki V,Xu L,Farah MH,Hatfield G,Kizana E,Marbán E,Koliatsos VEdoi
10.1016/j.jneumeth.2009.02.006subject
Has Abstractpub_date
2009-05-15 00:00:00pages
292-9issue
2eissn
0165-0270issn
1872-678Xpii
S0165-0270(09)00109-5journal_volume
179pub_type
杂志文章abstract：:This work demonstrates technical approaches to high-quality magnetic resonance imaging (MRI) of small structures of the mouse brain in vivo. It turns out that excellent soft-tissue contrast requires the reduction of partial volume effects by using 3D MRI at high (isotropic) resolution with linear voxel dimensions of a...
journal_title：Journal of neuroscience methods
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abstract：BACKGROUND:Advancement in clinically relevant studies like seizure interruption using functional neuro imaging tools has shown that specific changes in hemodynamics precede and accompany seizure onset and propagation. However, preclinical seizure experiments need to be conducted in awake animals with images reconstruct...
journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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doi：10.1016/j.jneumeth.2009.07.018
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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journal_title：Journal of neuroscience methods
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