Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

Research output: Contribution to journalJournal articleCommunication

Standard

Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets. / Griem-Krey, Nane; Klein, Anders Bue; Herth, Matthias; Wellendorph, Petrine.

In: Journal of visualized experiments : JoVE, No. 145, 12.03.2019.

Research output: Contribution to journalJournal articleCommunication

Harvard

Griem-Krey, N, Klein, AB, Herth, M & Wellendorph, P 2019, 'Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets' Journal of visualized experiments : JoVE, no. 145. https://doi.org/10.3791/58879

APA

Griem-Krey, N., Klein, A. B., Herth, M., & Wellendorph, P. (2019). Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets. Journal of visualized experiments : JoVE, (145). https://doi.org/10.3791/58879

Vancouver

Griem-Krey N, Klein AB, Herth M, Wellendorph P. Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets. Journal of visualized experiments : JoVE. 2019 Mar 12;(145). https://doi.org/10.3791/58879

Author

Griem-Krey, Nane ; Klein, Anders Bue ; Herth, Matthias ; Wellendorph, Petrine. / Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets. In: Journal of visualized experiments : JoVE. 2019 ; No. 145.

Bibtex

@article{dbb8093029244064b72441b836c4bd8e,
title = "Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets",
abstract = "In vitro autoradiography aims to visualize the anatomical distribution of a protein of interest in tissue from experimental animals as well as humans. The method is based on the specific binding of a radioligand to its biological target. Therefore, frozen tissue sections are incubated with radioligand solution, and the binding to the target is subsequently localized by the detection of radioactive decay, for example, by using photosensitive film or phosphor imaging plates. Resulting digital autoradiograms display remarkable spatial resolution, which enables quantification and localization of radioligand binding in distinct anatomical structures. Moreover, quantification allows for the pharmacological characterization of ligand affinity by means of dissociation constants (Kd), inhibition constants (Ki) as well as the density of binding sites (Bmax) in selected tissues. Thus, the method provides information about both target localization and ligand selectivity. Here, the technique is exemplified with autoradiographic characterization of the high-affinity γ-hydroxybutyric acid (GHB) binding sites in mammalian brain tissue, with special emphasis on methodological considerations regarding the binding assay parameters, the choice of the radioligand and the detection method.",
author = "Nane Griem-Krey and Klein, {Anders Bue} and Matthias Herth and Petrine Wellendorph",
year = "2019",
month = "3",
day = "12",
doi = "10.3791/58879",
language = "English",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "Journal of Visualized Experiments",
number = "145",

}

RIS

TY - JOUR

T1 - Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

AU - Griem-Krey, Nane

AU - Klein, Anders Bue

AU - Herth, Matthias

AU - Wellendorph, Petrine

PY - 2019/3/12

Y1 - 2019/3/12

N2 - In vitro autoradiography aims to visualize the anatomical distribution of a protein of interest in tissue from experimental animals as well as humans. The method is based on the specific binding of a radioligand to its biological target. Therefore, frozen tissue sections are incubated with radioligand solution, and the binding to the target is subsequently localized by the detection of radioactive decay, for example, by using photosensitive film or phosphor imaging plates. Resulting digital autoradiograms display remarkable spatial resolution, which enables quantification and localization of radioligand binding in distinct anatomical structures. Moreover, quantification allows for the pharmacological characterization of ligand affinity by means of dissociation constants (Kd), inhibition constants (Ki) as well as the density of binding sites (Bmax) in selected tissues. Thus, the method provides information about both target localization and ligand selectivity. Here, the technique is exemplified with autoradiographic characterization of the high-affinity γ-hydroxybutyric acid (GHB) binding sites in mammalian brain tissue, with special emphasis on methodological considerations regarding the binding assay parameters, the choice of the radioligand and the detection method.

AB - In vitro autoradiography aims to visualize the anatomical distribution of a protein of interest in tissue from experimental animals as well as humans. The method is based on the specific binding of a radioligand to its biological target. Therefore, frozen tissue sections are incubated with radioligand solution, and the binding to the target is subsequently localized by the detection of radioactive decay, for example, by using photosensitive film or phosphor imaging plates. Resulting digital autoradiograms display remarkable spatial resolution, which enables quantification and localization of radioligand binding in distinct anatomical structures. Moreover, quantification allows for the pharmacological characterization of ligand affinity by means of dissociation constants (Kd), inhibition constants (Ki) as well as the density of binding sites (Bmax) in selected tissues. Thus, the method provides information about both target localization and ligand selectivity. Here, the technique is exemplified with autoradiographic characterization of the high-affinity γ-hydroxybutyric acid (GHB) binding sites in mammalian brain tissue, with special emphasis on methodological considerations regarding the binding assay parameters, the choice of the radioligand and the detection method.

U2 - 10.3791/58879

DO - 10.3791/58879

M3 - Journal article

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 145

ER -

ID: 216920141