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Angiotensin II causes nitric oxide synthase 3 uncoupling in the thick ascending limb
Author(s) -
Massey Katherine,
Saikumar Jagannath J.,
Faber Mark D.,
Yee Jerry J.,
Garvin Jeffrey L.
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.lb807
Subject(s) - angiotensin ii , medicine , chemistry , nitric oxide , angiotensin ii receptor type 1 , endocrinology , receptor , nitric oxide synthase , basal (medicine) , biology , biochemistry , insulin
In the thick ascending limb (TAL) luminal flow increases NO production which inhibits flow‐mediated O 2 − production via a cGMP/PKG dependent pathway. Angiotensin II (Ang II) acutely stimulates TAL NO production by activating the AT2 receptor and stimulates O 2 − production via the AT1 receptor. We hypothesized that Ang II‐stimulated NO inhibits Ang II‐stimulated O 2 − via a cGMP‐dependent process. We measured NO production by fluorescence microscopy in isolated TAL. NO production was increased by 0.025 ± 0.004, 0.023 ± 0.014, 0.028 ± 0.007 arbitrary units (AU)/sec by 1, 10 and 100 pM Ang II, respesctively. Scavenging O 2 − with tempol did not affect Ang II‐stimulated NO production. We then measured O 2 − production in isolated TAL using a lucigenin‐based assay. Ang II (1 nM) increased O 2 − production by 1.77 ± 0.26 to 2.62 ± 0.36 AU ( p <0.01). Adding L‐arginine (L‐Arg), the substrate for NO production, had no effect on angiotensin II‐induced O 2 − . In the absence of L‐Arg the NOS inhibitor, L‐NAME, reduced Ang II‐stimulated O 2 − by 0.82 ± 0.31 AU ( p <0.05). In the presence of L‐Arg L‐NAME reduced O 2 − production by 0.71 ± 0.24 AU, however L‐NAME had no effect on basal O 2 − production. We conclude that: 1) Ang II increases both O 2 − production and NO production; 2) unlike flow‐induced NO, Ang II‐induced NO production does not inhibit Ang II‐induced O 2 − production; and 3) Ang II causes NOS uncoupling. Supported by NIH 5P01HL028982.