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NOS Antagonism Using Viral Vectors as an Experimental Strategy: Implications for In Vivo Studies of Cardiovascular Control and Peripheral Neuropathies

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Nitric Oxide

Part of the book series: Methods in Molecular Biology ((MIMB,volume 704))

Abstract

Nitric oxide, a free gaseous signalling molecule, has attracted the attention of numerous biologists and has been implicated in the regulation of the cardiovascular, nervous and immune system. However, the cellular mechanisms mediating nitric oxide modulation remain unclear. Upregulation by gene over-expression or down-regulation by gene inactivation of nitric oxide synthase has generated quantitative changes in abundance thereby permitting functional insights. We have tested and proved that genetic nitric oxide synthase antagonism using viral vectors, particularly with dominant negative mutants and microRNA 30-based short hairpin RNA , is an efficient and effective experimental approach to manipulate nitric oxide synthase expression both in vitro and in vivo.

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References

  1. Kasparov, S., Teschemacher, A. G., Hwang, D. Y., Kim, K. S., Lonergan, T., Paton, J. F. (2004) Viral vectors as tools for studies of central cardiovascular control. Prog Biophys Mol Biol 84, 251–277.

    Article  PubMed  CAS  Google Scholar 

  2. Aagaard, L., Rossi, J. J. (2007) RNAi therapeutics: principles, prospects and challenges. Adv Drug Deliv Rev 59, 75–86.

    Article  PubMed  CAS  Google Scholar 

  3. Thomas, C. E., Ehrhardt, A., Kay, M. A. (2003) Progress and problems with the use of viral vectors for gene therapy. Nat Rev Genet 4, 346–358.

    Article  PubMed  CAS  Google Scholar 

  4. Papale, A., Cerovic, M., Brambilla, R. (2009) Viral vector approaches to modify gene expression in the brain. J Neurosci Methods 15, 1–14.

    Article  Google Scholar 

  5. Snove, O., Jr., Rossi, J. J. (2006) Expressing short hairpin RNAs in vivo. Nat Methods 3, 689–695.

    Article  PubMed  CAS  Google Scholar 

  6. Paddison, P. J. (2008) RNA interference in mammalian cell systems. Curr Top Microbiol Immunol 320, 1–19.

    Article  PubMed  CAS  Google Scholar 

  7. Michel, T., Feron, O. (1997) Nitric oxide synthases: which, where, how, and why? J Clin Invest 100, 2146–2152.

    Article  PubMed  CAS  Google Scholar 

  8. Kantor, D. B., Lanzrein, M., Stary, S. J., Sandoval, G. M., Smith, W. B., Sullivan, B. M., et al. (1996) A role for endothelial NO synthase in LTP revealed by adenovirus-mediated inhibition and rescue. Science 274, 1744–1748.

    Article  PubMed  CAS  Google Scholar 

  9. Liu, J., Sessa, W. C. (1994) Identification of covalently bound amino-terminal myristic acid in endothelial nitric oxide synthase. J Biol Chem 269, 11691–11694.

    PubMed  CAS  Google Scholar 

  10. Busconi, L., Michel, T. (1993) Endothelial nitric oxide synthase. N-terminal myristoylation determines subcellular localization. J Biol Chem 268, 8410–8413.

    PubMed  CAS  Google Scholar 

  11. Harding, T. C., Geddes, B. J., Noel, J. D., Murphy, D., Uney, J. B. (1997) Tetracycline-regulated transgene expression in hippocampal neurones following transfection with adenoviral vectors. J Neurochem 69, 2620–2623.

    Article  PubMed  CAS  Google Scholar 

  12. Harding, T. C., Geddes, B. J., Murphy, D., Knight, D., Uney, J. B. (1998) Switching transgene expression in the brain using an adenoviral tetracycline-regulatable system. Nat Biotechnol 16, 553–555.

    Article  PubMed  CAS  Google Scholar 

  13. Wang, S., Teschemacher, A. G., Paton, J. F., Kasparov, S. (2006) Mechanism of nitric oxide action on inhibitory GABAergic signaling within the nucleus tractus solitarii. FASEB J 20, 1537–1539.

    Article  PubMed  CAS  Google Scholar 

  14. Waki, H., Murphy, D., Yao, S. T., Kasparov, S., Paton, J. F. (2006) Endothelial NO synthase activity in nucleus tractus solitarii contributes to hypertension in spontaneously hypertensive rats. Hypertension 48, 644–650.

    Article  PubMed  CAS  Google Scholar 

  15. Waki, H., Kasparov, S., Wong, L. F., Murphy, D., Shimizu, T., Paton, J. F. (2003) Chronic inhibition of endothelial nitric oxide synthase activity in nucleus tractus solitarii enhances baroreceptor reflex in conscious rats. J Physiol 546, 233–242.

    Article  PubMed  CAS  Google Scholar 

  16. Paton, J. F., Waki, H., Abdala, A. P., Dickinson, J.,, Kasparov, S. (2007) Vascular-brain signaling in hypertension: role of angiotensin II and nitric oxide. Curr Hypertens Rep 9, 242–247.

    Article  PubMed  CAS  Google Scholar 

  17. Bridge, P. M., Ball, D. J., Mackinnon, S. E., Nakao, Y., Brandt, K., Hunter, D. A., et al. (1994) Nerve crush injuries – a model for axonotmesis. Exp Neurol 127, 284–290.

    Article  PubMed  CAS  Google Scholar 

  18. Verdu, E., Ceballos, D., Vilches, J. J., Navarro, X. (2000) Influence of aging on peripheral nerve function and regeneration. J Peripher Nerv Syst 5, 191–208.

    Article  PubMed  CAS  Google Scholar 

  19. Gonzalez-Hernandez, T., Rustioni, A. (1999) Expression of three forms of nitric oxide synthase in peripheral nerve regeneration. J Neurosci Res 55, 198–207.

    Article  PubMed  CAS  Google Scholar 

  20. Sunico, C. R., Portillo, F., Gonzalez-Forero, D., Kasparov, S.,, Moreno-Lopez, B. (2008) Evidence for a detrimental role of nitric oxide synthesized by endothelial nitric oxide synthase after peripheral nerve injury. Neuroscience 157, 40–51.

    Article  PubMed  CAS  Google Scholar 

  21. Stegmeier, F., Hu, G., Rickles, R. J., Hannon, G. J., Elledge, S. J. (2005) A lentiviral microRNA-based system for single-copy polymerase II-regulated RNA interference in mammalian cells. Proc Natl Acad Sci USA 102, 13212–13217.

    Article  PubMed  CAS  Google Scholar 

  22. Liu, B., Paton, J. F., Kasparov, S. (2008) Viral vectors based on bidirectional cell-specific mammalian promoters and transcriptional amplification strategy for use in vitro and in vivo. BMC Biotechnol 8, 49–57.

    Article  PubMed  Google Scholar 

  23. Sunico, C. R., Portillo, F., Gonzalez-Forero, D., Moreno-Lopez, B. (2005) Nitric-oxide-directed synaptic remodeling in the adult mammal CNS. J Neurosci 25, 1448–1458.

    Article  PubMed  CAS  Google Scholar 

  24. Coleman, J. E., Huentelman, M. J., Kasparov, S., Metcalfe, B. L., Paton, J. F., Katovich, M. J., et al. (2003) Efficient large-scale production and concentration of HIV-1-based lentiviral vectors for use in vivo. Physiol Genomics 12, 221–228.

    PubMed  CAS  Google Scholar 

  25. Bewig, B., Schmidt, W. E. (2000) Accelerated titering of adenoviruses. Biotechniques 28, 870–873.

    PubMed  CAS  Google Scholar 

  26. Lonergan, T., Teschemacher, A. G., Hwang, D. Y., Kim, K. S., Pickering, A. E., Kasparov, S. (2005) Targeting brain stem centers of cardiovascular control using adenoviral vectors: impact of promoters on transgene expression. Physiol Genomics 20, 165–172.

    Article  PubMed  CAS  Google Scholar 

  27. Gonzalez-Forero, D., Portillo, F., Sunico, C. R., Moreno-Lopez, B. (2004) Nerve injury reduces responses of hypoglossal motoneurones to baseline and chemoreceptor-modulated inspiratory drive in the adult rat. J Physiol 557, 991–1011.

    Article  PubMed  CAS  Google Scholar 

  28. Xu, H., McCann, M., Zhang, Z., Posner, G. H., Bingham, V., El-Tanani, M., et al. (2009) Vitamin D receptor modulates the neoplastic phenotype through antagonistic growth regulatory signals. Mol Carcinog 48, 758–772.

    Article  PubMed  CAS  Google Scholar 

  29. Chen, X., Xu, H., Wan, C., McCaigue, M., Li, G. (2006) Bioreactor expansion of human adult bone marrow-derived mesenchymal stem cells. Stem Cells 24, 2052–2059.

    Article  PubMed  CAS  Google Scholar 

  30. Paxinos, G., Watson, C. (1986) The Rat Brain in Stereotaxic Coordinates. Academic, London.

    Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Physiology and Pharmacology, University of Bristol, Bristol, UK

    Beihui Liu, James Hewinson, Julian F.R. Paton & Sergey Kasparov

  2. Department of Pharmacology, State Key Laboratory for Research and Development of Chinese Materia Medica, Chengdu University of Traditional Chinese Medicine, Chengdu, P.R. China

    Haibo Xu

  3. Área de Fisiología, Facultad de Medicina, Universidad de Cádiz, Cádiz, Spain

    Francisco Montero, Carmen R. Sunico, Federico Portillo & Bernardo Moreno-López

Authors
  1. Beihui Liu
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  2. James Hewinson
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  3. Haibo Xu
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  4. Francisco Montero
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  5. Carmen R. Sunico
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  6. Federico Portillo
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  7. Julian F.R. Paton
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  8. Bernardo Moreno-López
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  9. Sergey Kasparov
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Corresponding author

Correspondence to Beihui Liu .

Editor information

Editors and Affiliations

  1. School of Pharmacy, Queen's University, Belfast, Lisburn Road 97, Belfast, BT9 7BL, United Kingdom

    Helen O. McCarthy

  2. School of Pharmacy, Queen's University, Belfast, Lisburn Road 97, Belfast, BT9 7BL, United Kingdom

    Jonathan A. Coulter

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Liu, B. et al. (2011). NOS Antagonism Using Viral Vectors as an Experimental Strategy: Implications for In Vivo Studies of Cardiovascular Control and Peripheral Neuropathies. In: McCarthy, H., Coulter, J. (eds) Nitric Oxide. Methods in Molecular Biology, vol 704. Humana Press. https://doi.org/10.1007/978-1-61737-964-2_15

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  • DOI: https://doi.org/10.1007/978-1-61737-964-2_15

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61737-963-5

  • Online ISBN: 978-1-61737-964-2

  • eBook Packages: Springer Protocols

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