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’).

AT1a/GFP/HEK cell recording

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.

AT1a/GFP/HEK cells

[K307Q]AT1a/GFP/HEK cells