title: Transgenic mice expressing an intracellular fluorescent fusion of angiotensin II demonstrate renal thrombotic microangiopathy and elevated blood pressure.
abstract: We have generated transgenic mice that express angiotensin II (ANG II) fused downstream of enhanced cyan fluorescent protein, expression of which is regulated by the mouse metallothionein promoter. The fusion protein, which lacks a secretory signal, is retained intracellularly. In the present study, RT-PCR, immunoblot analyses, whole-animal fluorescent imaging, and fluorescent microscopy of murine embryonic fibroblasts confirm expression of the fusion protein in vivo and in vitro. The transgene is expressed in all tissues tested (including brain, heart, kidney, liver, lung, and testes), and radioimmunoassay of plasma samples obtained from transgenic mice indicate no increase in circulating ANG II over wild-type levels, consistent with intracellular retention of the transgene product. Kidneys from transgenic and corresponding wild-type littermates were histologically evaluated, and abnormalities in transgenic mice consistent with thrombotic microangiopathy were observed; microthrombosis was frequently observed within the glomerular capillaries and small vessels. In addition, systolic and diastolic blood pressures, measured by telemetry (n = 8 for each group), were significantly higher in transgenic mice compared with wild-type littermates. Blood pressure of line A male transgenic mice was 125 + or - 1.7 over 97 + or - 1.6 compared with 109 + or - 1.7 over 83 + or - 1.4 mmHg in wild-type littermates (systolic over diastolic). In summary, overexpression of an intracellular fluorescent fusion protein of ANG II correlates with elevated blood pressure and kidney pathology. This transgenic model may be useful to further explore the intracellular renin-angiotensin system and its implication in abnormal kidney function and hypertension.
authors.affiliation: Department of Molecular Genetics, Ochsner Clinic Foundation, 1514 Jefferson Highway, Biomedical Research Bldg., New Orleans, LA 70121, USA.
authors.forename: K M
authors.initials: KM
authors.lastname: Redding
authors.forename: B L
authors.initials: BL
authors.lastname: Chen
authors.forename: A
authors.initials: A
authors.lastname: Singh
authors.forename: R N
authors.initials: RN
authors.lastname: Re
authors.forename: L G
authors.initials: LG
authors.lastname: Navar
authors.forename: D M
authors.initials: DM
authors.lastname: Seth
authors.forename: C D
authors.initials: CD
authors.lastname: Sigmund
authors.forename: W W
authors.initials: WW
authors.lastname: Tang
authors.forename: J L
authors.initials: JL
authors.lastname: Cook
mesh_terms: D000804:Angiotensin II / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001794:Blood Pressure / Q000502:physiology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006973:Hypertension / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D007668:Kidney / Q000098:blood supply* / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D012333:RNA, Messenger / Q000378:metabolism; D057049:Thrombotic Microangiopathies / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology
chemical_list: D012333:RNA, Messenger; C527701:enhanced cyan fluorescent protein; D000804:Angiotensin II; D049452:Green Fluorescent Proteins