Logo image
Back
Demonstration of nuclear translocation of the mineralocorticoid receptor (MR) using an anti-MR antibody and confocal laser scanning microscopy
Journal article   Open access   Peer reviewed

Demonstration of nuclear translocation of the mineralocorticoid receptor (MR) using an anti-MR antibody and confocal laser scanning microscopy

N M Robertson, G Schulman, S Karnik, E Alnemri and G Litwack
Molecular endocrinology (Baltimore, Md.), v 7(9), pp 1226-1239
01 Sep 1993
DOI: https://doi.org/10.1210/me.7.9.1226
PMID: 8247024
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.1210/me.7.9.1226View
Published, Version of Record (VoR) Open

Abstract

Amino Acid Sequence Animals Antibodies Blotting, Western Carcinoma, Hepatocellular - metabolism Carcinoma, Hepatocellular - pathology Cell Nucleus - metabolism Cross Reactions Cytosol - metabolism Fluorescent Antibody Technique Humans Immunohistochemistry Kidney - cytology Kidney - metabolism Kidney Cortex - cytology Kidney Cortex - metabolism Liver Neoplasms - metabolism Liver Neoplasms - pathology Macrophages - cytology Macrophages - metabolism Mice Microscopy, Fluorescence - methods Molecular Sequence Data Oligopeptides - immunology Rats Receptors, Mineralocorticoid - analysis Receptors, Mineralocorticoid - immunology Receptors, Mineralocorticoid - metabolism
Using a synthetic peptide that corresponds to a unique region of the N-terminal domain of the human mineralocorticoid receptor (MR), amino acids 87-96, we have generated a polyclonal antibody, human (h) MRsN. This sequence shares no homology with the corresponding sequences of the glucocorticoid receptor or other steroid/thyroid hormone receptor superfamily members. Antibody hMRsN cross-reacts with MR from human, rat, and mouse cells and recognizes denatured MR from either crude preparations or partially purified rat kidney cytosol, rat colon, or recombinant hMR overexpressed in baculovirus-infected Sf9 cells. Immunoprecipitation of the native MR from either partially purified or crude preparations of rat kidney cytosol with hMRsN, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver stain, demonstrated a major protein band with a mol wt of 116 kilodaltons. In addition, using confocal laser scanning microscopy and digital image analysis, the immunocytochemical localization of the recombinant hMR over-expressed in Sf9 cells 24 h post-transfection was determined. In the absence of ligand, the MR was detected solely in the cytoplasm, after a 30-min exposure to 100 nM aldosterone the MR was perinuclear, and after 60 min, the MR was predominantly nuclear. To ascertain that this phenomenon was not unique to insect cells, aldosterone induced MR nuclear translocation in mouse macrophage cells was also demonstrated immunocytochemically, clearly indicating a role for nuclear translocation in MR function.

Metrics

5 Record Views
83 citations in Web of Science

Details

UN Sustainable Development Goals (SDGs)

This publication has contributed to the advancement of the following goals:

#3 Good Health and Well-Being

InCites Highlights

Data related to this publication, from InCites Benchmarking & Analytics tool:

Collaboration types
Domestic collaboration
Web of Science research areas
Endocrinology & Metabolism
Logo image