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Long‐term effect of neuronal nitric oxide synthase over‐expression on cardiac neurotransmission mediated by a lentiviral vector
Author(s) -
Wang Lijun,
Li Dan,
Dawson Tom A.,
Paterson David J.
Publication year - 2009
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2009.172866
Subject(s) - tyrosine hydroxylase , choline acetyltransferase , neurotransmission , neuromodulation , nitric oxide synthase , cholinergic , acetylcholine , chemistry , endocrinology , medicine , gene expression , microbiology and biotechnology , biology , nitric oxide , stimulation , dopamine , receptor , gene , biochemistry
Short‐term over‐expression of neuronal nitric oxide synthase (nNOS) with adenoviral gene transfer into peripheral cardiac autonomic neurons can facilitate cholinergic neurotransmission, and inhibit sympathetic transmission, by regulating cyclic nucleotide‐dependent pathways coupled to neuronal calcium entry. We tested the idea whether cardiac neuromodulation by nNOS could be sustained by long‐term over‐expression of the enzyme following lentiviral gene transfer. We developed a lentiviral vector with an elongation factor 1 (EF1α) promoter to drive nNOS or enhanced green fluorescent protein (eGFP) expression. Lenti.EF1α‐nNOS or Lenti.EF1α‐eGFP was transferred to the right atrium of Spague–Dawley (SD) rats and acetylcholine (ACh) or noradrenaline (NA) release to field stimulation was measured 4 months after gene transfer. Atria transduced with Lenti.EF1α‐nNOS had higher nNOS expression compared to the atria treated with Lenti.EF1α‐eGFP ( P < 0.05). We also detected significant increases ( P < 0.05) in atrial cGMP and cAMP levels in the same tissue. Immunohistochemistry revealed co‐localisation of eGFP in intrinsic cholinergic neurons (choline acetyltransferase positive) and intrinsic adrenergic neurons (tyrosine hydroxylase positive) following gene transfer. nNOS‐transduced animals displayed enhanced ACh release ( P < 0.05) and reduced NA release ( P < 0.05) compared to the eGFP‐treated group. nNOS‐specific inhibition reversed the enhanced ACh release. Persistent nNOS over‐expression mediated by a lentiviral vector can modulate sympatho‐vagal control of cardiac excitability. This approach may provide a new tool to target impaired cardiac autonomic phenotypes that are disrupted by several cardiovascular pathologies.