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Manipulation of neuronal nitric oxide synthase within the paraventricular nucleus using adenovirus and antisense technology.
Methods Mol Med. 2005; 112:59-79.MM

Abstract

Congestive heart failure (CHF) is characterized by impaired cardiovascular reflexes and increased neurohumoral drive. The long-term sympatho-excitation increases the progression and risk of mortality during CHF. The paraventricular nucleus (PVN) of the hypothalamus is a very important central site for integration of sympathetic outflow and cardiovascular function. Within the PVN, nitric oxide (NO), mainly generated by neuronal nitric oxide synthase (nNOS), functions in inhibitory regulation of sympathetic outflow. Our previous study has indicated that in rats with experimental heart failure, the NO mechanism within the PVN is attenuated. We hypothesize that this alteration may contribute to the sympatho-excitation commonly observed in CHF. To investigate the role of NO within the PVN in sympathetic dysfunction in CHF, we have manipulated nNOS expression using adenoviral gene transfer of nNOS or nNOS antisense. These techniques have allowed us to observe the effects of alterations in nNOS on sympathetic outflow and cardiovascular function. In this chapter, we describe the methods for delivering nNOS adenoviral vector or nNOS antisense into the PVN using microinjection, as well as the protocols for detecting nNOS expression after these manipulations, using Western blot, NADPH-diaphorase staining, and immunofluorescent staining.

Authors+Show Affiliations

Department of Physiology and Biophysics, University of Nebraska Medical Center, Omaha, NE, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Review

Language

eng

PubMed ID

16010011

Citation

Li, Yi-Fan, et al. "Manipulation of Neuronal Nitric Oxide Synthase Within the Paraventricular Nucleus Using Adenovirus and Antisense Technology." Methods in Molecular Medicine, vol. 112, 2005, pp. 59-79.
Li YF, Wang Y, Channon KM, et al. Manipulation of neuronal nitric oxide synthase within the paraventricular nucleus using adenovirus and antisense technology. Methods Mol Med. 2005;112:59-79.
Li, Y. F., Wang, Y., Channon, K. M., Schultz, H. D., Zucker, I. H., & Patel, K. P. (2005). Manipulation of neuronal nitric oxide synthase within the paraventricular nucleus using adenovirus and antisense technology. Methods in Molecular Medicine, 112, 59-79.
Li YF, et al. Manipulation of Neuronal Nitric Oxide Synthase Within the Paraventricular Nucleus Using Adenovirus and Antisense Technology. Methods Mol Med. 2005;112:59-79. PubMed PMID: 16010011.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Manipulation of neuronal nitric oxide synthase within the paraventricular nucleus using adenovirus and antisense technology. AU - Li,Yi-Fan, AU - Wang,Yu, AU - Channon,Keith M, AU - Schultz,Harold D, AU - Zucker,Irving H, AU - Patel,Kaushik P, PY - 2005/7/13/pubmed PY - 2005/8/5/medline PY - 2005/7/13/entrez SP - 59 EP - 79 JF - Methods in molecular medicine JO - Methods Mol Med VL - 112 N2 - Congestive heart failure (CHF) is characterized by impaired cardiovascular reflexes and increased neurohumoral drive. The long-term sympatho-excitation increases the progression and risk of mortality during CHF. The paraventricular nucleus (PVN) of the hypothalamus is a very important central site for integration of sympathetic outflow and cardiovascular function. Within the PVN, nitric oxide (NO), mainly generated by neuronal nitric oxide synthase (nNOS), functions in inhibitory regulation of sympathetic outflow. Our previous study has indicated that in rats with experimental heart failure, the NO mechanism within the PVN is attenuated. We hypothesize that this alteration may contribute to the sympatho-excitation commonly observed in CHF. To investigate the role of NO within the PVN in sympathetic dysfunction in CHF, we have manipulated nNOS expression using adenoviral gene transfer of nNOS or nNOS antisense. These techniques have allowed us to observe the effects of alterations in nNOS on sympathetic outflow and cardiovascular function. In this chapter, we describe the methods for delivering nNOS adenoviral vector or nNOS antisense into the PVN using microinjection, as well as the protocols for detecting nNOS expression after these manipulations, using Western blot, NADPH-diaphorase staining, and immunofluorescent staining. SN - 1543-1894 UR - https://www.unboundmedicine.com/medline/citation/16010011/Manipulation_of_neuronal_nitric_oxide_synthase_within_the_paraventricular_nucleus_using_adenovirus_and_antisense_technology_ DB - PRIME DP - Unbound Medicine ER -