Technical Reference #1662

1662.

Improvement of Renal Hemodynamics During Hypertension-Induced Chronic Renal Disease: Role of EGF Receptor Antagonism Frank Helle; Charlotte Jouzel; Christos Chadjichristos; Sandrine Placier; Martin Flamant; Dominique Guerrot; He´le`ne Franc¸ois; Jean-Claude Dussaule;and Christos Chatziantoniou, INSERM, American Journal of Physiology: Renal Physiology, 297(1662), (2009)
Link To Paper

Abstract:
The present study investigated mechanisms of regression of renal disease after severe proteinuria byfocusing on the interaction among EGF receptors renal hemodynamicsand structural lesions.

Keywords:
angiotensin; fibrosis; proteinuria; regression

Materials & Methods:
Mean lumen diameter and afferent arteriole hypertrophic measurements. Isolated vessels were attached by self-adhesion to the clean glass surface of a no. 0 coverslip mounted in a petri dish (MatTek P35G-0-14-C) containing 3 ml medium without Ca2 . Before the experiments Ca2 concentration in the medium was gradually increased to 2 mmol/l in three steps (20 mol/l 200 mol/l and 2 mmol/l) with a 3- to 5-min incubation in between. To stimulate the vessels 300 l medium was aspirated from the dish and the same volume containing 10 8 mol/l endothelin-1 was gently superfused ( 100 l/s) over the vessels using a 1-ml pipette (Gilson) producing a final concentration of 10 9 mol/l endothelin-1 in the vessel bath. Brightfield images (Leica SP2 camera with 40 objective 512 512 pixels field of view 480 480 m2 optical resolution 0.9 m) were acquired at baseline and every 30 s for 6 min after addition of the agonist. The inner outline of the vessel wall was traced using Olympus DP-Soft 5.0 software. Mean lumen diameter was calculated using the formula mean lumen diameter lumen area/lumen length. Mean vessel wall thickness was calculated using the formula mean vessel wall [(arteriole area lumen area)/2]/arteriole length. The media/ lumen ratio was calculated as mean vessel wall thickness/mean lumen diameter. Up to two arterioles from each kidney were used for contractility experiments and 10–17 arterioles were used in each experimental group. For measurements of afferent arteriole vessel wall thickness and media/lumen ratio 1–3 arterioles were harvested from each kidney and 17–28 arterioles were used in each experimental group.

Microscopic Technique
NA

Cell Type(s)
Arteriole