Document Detail


Visualization of binding sites for bovine parathyroid hormone (PTH 1-84) on cultured kidney cells with a biotinyl-b-PTH (1-84) antagonist.
MedLine Citation:
PMID:  3512693     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Parathyroid hormone (PTH) receptors have been found in a subpopulation of kidney cells. In this report, we investigated the feasibility of techniques that apply a partial antagonist of PTH conjugated to biotin to localize receptors cytochemically on bovine kidney cortical cells in monolayer culture at the light microscopic level. Biotinylated bovine PTH (1-84) (biotinyl-PTH) was bound to the cultured cells for 1-30 min at 37 degrees C in the amounts of 10(-5) -10(-10) M. In a different set of experiments, the cells were also exposed to a solution containing 10(-6) M biotinylated PTH and an excess of unlabeled PTH, insulin, adrenocorticotropin, or calcitonin for 10 and 30 min at 37 degrees C to test the specificity of the binding. The cells were then fixed in 2.5% glutaraldehyde and stained with the avidin-biotin peroxidase complex (ABC) technique. Diffuse labeling was evident on 30% of the cells in 10 min with concentrations of biotinyl-PTH as low as 10(-8) M. The stain was diffuse, but more intense after 1-10 min in higher concentrations (10(-6) M). If a 15-1500-fold excess of unlabeled PTH was added to the biotinyl-PTH, no staining was observed. The other peptides (insulin, ACTH or calcitonin) had no effect on binding. Longer times in biotinyl-PTH (10(-6) M for 10-30 min) resulted in intense patches of label on the cells resembling caps (in addition to the pale diffuse label). The percentage of labeled cells in the monolayer (30%) did not change with time. These studies show that a partial antagonist of PTH can be used as a cytochemical probe for specific PTH receptors in a subpopulation of cultured cortical kidney cells.
Authors:
A Niendorf; M Dietel; H Arps; J Lloyd; G V Childs
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society     Volume:  34     ISSN:  0022-1554     ISO Abbreviation:  J. Histochem. Cytochem.     Publication Date:  1986 Mar 
Date Detail:
Created Date:  1986-04-01     Completed Date:  1986-04-01     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9815334     Medline TA:  J Histochem Cytochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  357-61     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Binding Sites
Biotin / analogs & derivatives*,  pharmacology
Cattle
Cell Survival
Cells, Cultured
Histocytochemistry / methods
Kidney / cytology,  metabolism*
Kidney Cortex / cytology,  metabolism
Microscopy, Interference
Microscopy, Phase-Contrast
Parathyroid Hormone / antagonists & inhibitors,  metabolism*,  pharmacology*
Time Factors
Chemical
Reg. No./Substance:
0/Parathyroid Hormone; 0/biotinyl-parathyroid hormone; 58-85-5/Biotin

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


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