ANGIOTENSIN II AT1a RECEPTOR TRANSLOCATION TO THE NUCLEUS
Tom Morinelli, Ph.D.
Ralph H. Johnson VA Medical Research Center,
Division of Nephrology, Department of Medicine,
Medical University of South Carolina
Charleston, SC
Time
lapse real time recordings of live cells exposed to AngII (100nM). Nuclei are
stained with DRAQ5. Images
acquired with the PerkinElmer UltraView system using a 40x oil objective. Cells maintained in Hank’s Balanced
Salt Solution, pH 7.4, 37°C for the duration of the study (30’-60’).
HEK-293
cells expressing the Wild-type receptor (AT1a/GFP) respond by morphological shape changes followed by receptor
internalization and trafficking to the nuclear area.
[STL335-338AAA]AT1a/GFP/HEK
cell recording.
This
shows the internalization-deficient mutant receptor expressing cells responding
to the addition of AngII by morphological shape changes without subsequent
internalization of the receptor nor nuclear localization.
[K307Q]AT1a/GFP/HEK
cell recording.
This
recording shows NLS-deficient mutant receptor expressing cells responding to
AngII by morphological shape changes, receptor internalization but without
apparent nuclear localization of the receptor.
LSCM 3-D renderings of Human Embryonic Kidney (HEK) 293
Cells expressing the wild type AT1a/GFP construct and the NLS-deficient mutant
[K307Q]/AT1a/GFP after stimulation by AngII (100nM, 60 min). Image rendering performed using
Velocity Software (Improvision) on z-axis scans. DRAQ5 (Alexis Corp., San Diego, CA) used to stain nuclear
DNA.