The examination and quantitation of tissue cytosolic receptors for 2,3,7,8-tetrachlorodibenzo-p-dioxin using hydroxylapatite

Journal Article · Thu Jul 15 00:00:00 EDT 1982 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(82)90212-3· OSTI ID:6324183

Gasiewicz, T A ^[1]; Neal, R A

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1. Univ. of Rochester, NY

An assay using hydroxylapatite has been developed for the examination and quantitation of cytosolic receptors for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). This method, as compared to others, has a relatively high ratio of specific to nonspecific binding of (/sup 3/H)TCDD and is relatively rapid. The total number of specific binding sites and equilibrium dissociation constants (K/sub D/) for (/sup 3/H)TCDD were determined in hepatic cytosol from Sprague-Dawley rats and C57BL/6J, DBA/2J, and B6D2F/sub 1//J mice. With the exception of the cytosol from DBA/2J mice in which little specific binding was observed, high-affinity, specific binding was obtained in all cases. The rat hepatic cytosol demonstrated the highest affinity for (/sup 3/H)TCDD with a K/sub D/ of 0.12 nM. The total amount of specific (/sup 3/H)TCDD binding in hepatic cytosol from B6D2F/sub 1//J mice was found to be intermediate between that of the C57BL/6J and DBA/2J parents. Ligand competition studies suggest the specificity of binding of various compounds to the receptor species from Sprague-Dawley rats is similar to that observed with the hepatic cytosol from C57BL/6J mice. These data support the hypothesis that, as in mice, the particular cytosol binding species in rats is the receptor that may mediate the induction of aryl hydrocarbon hydroxylase activity.

• DOE Contract Number: AC02-76EV03490
• OSTI ID: 6324183
• Journal Information: Anal. Biochem.; (United States), Vol. 124:1
• Country of Publication: United States
• Language: English

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