Document Detail


Mineralocorticoid and glucocorticoid receptor steroid binding and localization in colonic cells.
MedLine Citation:
PMID:  8166236     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In rat colon epithelium glucocorticoids and mineralocorticoids regulate Na transport by binding to distinct receptors and stimulating different pathways. The distribution and intracellular localization of mineralocorticoid (MR) and glucocorticoid (GR) receptors in colonic Na-absorbing surface cells and Cl-secreting crypt cells is unknown. Surface and crypt cells were sequentially isolated from rat distal colon by EDTA chelation and mechanical dissociation. Cell viability was confirmed by trypan blue exclusion and low rates of 2',7'-bis(2-carboxyethyl)-5(6)-carboxylfluorescein leak. Histologic examination, alkaline phosphatase activity, and rates of [3H]leucine incorporation confirmed separation of surface from crypt cells. Scatchard analysis of [3H]aldosterone and [3H]triamcinolone acetonide binding demonstrated that the number of MR decreased from 7,228 +/- 1,067 in surface to 2,299 +/- 434 receptors/cell in crypt cells, whereas the number of GR increased from 20,857 +/- 4,241 in surface to 58,598 +/- 8,207 receptors/cell in crypt cells. The dissociation constants were 2.8 +/- 0.4 nM for the MR and 12 +/- 3 nM for the GR. Indirect immunofluorescence using the specific anti-MR antibody hMRsN and the anti-GR antibody BuGR-2 demonstrated that both unliganded receptors were cytoplasmic and translocated to the nucleus after hormone binding. These data indicate that both surface and crypt cells are potentially responsive to mineralocorticoids and glucocorticoids and that both the MR and GR require hormone for nuclear translocation.
Authors:
G Schulman; N M Robertson; I B Elfenbein; D Eneanya; G Litwack; C P Bastl
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The American journal of physiology     Volume:  266     ISSN:  0002-9513     ISO Abbreviation:  Am. J. Physiol.     Publication Date:  1994 Mar 
Date Detail:
Created Date:  1994-05-20     Completed Date:  1994-05-20     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370511     Medline TA:  Am J Physiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  C729-40     Citation Subset:  IM    
Affiliation:
Department of Medicine, Temple University Health Sciences Center, Philadelphia 19140.
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MeSH Terms
Descriptor/Qualifier:
Aldosterone / metabolism*
Animals
Colon / cytology,  metabolism*
Fluoresceins
Fluorescent Antibody Technique
Fluorescent Dyes
Rats
Rats, Sprague-Dawley
Receptors, Glucocorticoid / metabolism*
Receptors, Mineralocorticoid / metabolism*
Tissue Distribution
Triamcinolone Acetonide / metabolism*
Grant Support
ID/Acronym/Agency:
R01-DK-27486/DK/NIDDK NIH HHS; S07RR05417/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Fluoresceins; 0/Fluorescent Dyes; 0/Receptors, Glucocorticoid; 0/Receptors, Mineralocorticoid; 52-39-1/Aldosterone; 76-25-5/Triamcinolone Acetonide; 85138-49-4/2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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