Shear stress affects the intracellular distribution of eNOS: direct demonstration by a novel in vivo technique.
The focal location of atherosclerosis in the vascular tree is correlated with local variations in shear stress. We developed a method to induce defined variations in shear stress in a straight vessel segment of a mouse. To this end, a cylinder with a tapered lumen was placed around the carotid artery, inducing a high shear stress field. Concomitantly, regions of low shear stress and oscillatory shear stress were created upstream and down-stream of the device, respectively. This device was used in mice transgenic for an eNOS3GFP fusion gene. We observed a strong induction of endothelial nitric oxide synthase-green fluorescent protein (eNOS-GFP) mRNA expression in the high shear stress region compared with the other regions (P < .05). Quantification of eNOS-GFP fluorescence or of immunoreactivity to the Golgi complex or to platelet endothelial cell adhesion molecule 1 (PECAM-1) showed an increase in the high shear stress region (P < .05) compared with nontreated carotid arteries. Colocalization of eNOS-GFP with either the Golgi complex or PECAM-1 also responded to alterations of shear stress. In conclusion, we showed a direct response of mRNA and protein expression in vivo to induced variations of shear stress. This model provides the opportunity to study the relationship between shear stress alterations, gene expression, and atherosclerosis.
Cheng
Caroline
C
Department of Cardiology, Erasmus University Medical Center, Rotterdam, The Netherlands.
van Haperen
Rien
R
de Waard
Monique
M
van Damme
Luc C A
LC
Tempel
Dennie
D
Hanemaaijer
Laurens
L
van Cappellen
Gert W A
GW
Bos
Joop
J
Slager
Cornelis J
CJ
Duncker
Dirk J
DJ
van der Steen
Anton F W
AF
de Crom
Rini
R
Krams
Rob
R
D000818:Animals; D050197:Atherosclerosis / Q000503:physiopathology; D002339:Carotid Arteries / Q000201:enzymology*; D005786:Gene Expression Regulation; D006056:Golgi Apparatus / Q000378:metabolism; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D016133:Polymerase Chain Reaction; D021381:Protein Transport / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D033081:Shear Strength; D013314:Stress, Mechanical; D018615:Ultrasonography, Doppler, Color
D019408:Platelet Endothelial Cell Adhesion Molecule-1; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse