Imaging receptors by autoradiography: Computer-assisted approaches

Kuhar, Michael J.; Lloyd, D.; Appel, Nathan M.; Loats, H.L.

doi:10.1016/0891-0618(91)90040-j

Cited by 14 publications

(4 citation statements)

References 18 publications

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“…Emulsion-covered coverslips and subsequent microscopic examination have been widely used, especially in the initial studies. The introduction of tritium-sensitive sheet films for the visualization of the labeled receptors (Palacios et al, 1981) represented an important advance because of their ease of use and, more importantly, because they allow accurate, rapid, and relatively easy quantification of the densities of receptors with the help of computer-assisted image analysis systems (Kuhar et al, 1991 ;Palacios et al, 1991).…”

Section: Methods For the Visualization O F Neurotransmitter Receptorsmentioning

confidence: 99%

Receptor distribution in the human and animal hippocampus: Focus on muscarinic acetylcholine receptors

et al. 1993

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Section: Methods For the Visualization O F Neurotransmitter Receptorsmentioning

confidence: 99%

Receptor distribution in the human and animal hippocampus: Focus on muscarinic acetylcholine receptors

et al. 1993

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“…Thus, [ 3H]-esuadiol and [ 3HJ-moxestrol both give good results, albeit after a 3-month exposure as opposed to the 1-day exposure required with [ 12'I]-MIE2. Non-photographic image-enhancement techniques Kuhar et al, 1991) may provide a valuable adjunct to studies with 3H-labeled ligands to achieve shorter exposure times. Results with the commercially available iba-[ '2'I]-iodo-estradiol, by contrast, were not satisfactory.…”

Section: Discussionmentioning

confidence: 99%

In vitro autoradiographic visualization of occupied estrogen receptors in the rat brain with an iodinated estrogen ligand.

Walters¹,

Brown²,

Hochberg³

et al. 1993

J Histochem Cytochem.

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Methods have been developed for the selective measurement of occupied estrogen receptors (ER) in brain tissue sections.Cryostat sections of d i e d tissue were incubated with radiolabeled estrogen at physiological temperatures, displacing endogenous receptot-bound estrogen by radioligand and thereby allowing the receptor complexes to be visualized autoradiographically after washing to remove nonspecifically bound steroid. The resultant autoradiographs were analyzed by computer-assisted densitometry. Synthetic 1 lpmethoxysubstituted radiolabeled estrogens gave the best autoradiographic images, as a result of reduced nonspecific labeling, although [3H]-estradiol was also used successfully. With the attoduction The actions of steroids on the brain involve effects on many different cell groups, each with its own specific populations of steroidsensitive and steroid-insensitive cells and regional patterns of metabolism. This complexity and heterogeneity make it difficult to elucidate the actions of steroids through measurements performed either on the whole brain or on crudely dissected neural structures. Methodological difficulties in the measurement of steroid receptors in small tissue samples have, however, hindered attempts to more precisely define regional patterns of steroid receptor occupation under physiological conditions of steroid hormone secretion.

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“…During the last decade, autoradiograms on film or micro-autoradiograms on histological sections have been quantified using image analysers calibrated with known standards (Kuhar et al, 1991). The determination of receptor specificity and affinity became possible on very small structures of interest using quantitative *To whom correspondence should be addressed.…”

Section: Introductionmentioning

confidence: 99%

Radioimagers as an alternative to film autoradiography forin situ quantitative analysis of125I-ligand receptor binding and pharmacological studies

et al. 1996

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Three radioimagers, the mu-imager, the beta-imager and the phosphorimager, were tested as alternatives to quantitative autoradiography on film, for receptor imaging and pharmacological in situ quantitative analysis. Two iodinated ligands 125I-interleukin-1 alpha and 125I-gonadotropin releasing hormone agonist were used for receptor characterization in mouse brain and pituitary sections. Due to the high number of the agonist receptors in rat pituitary gland, this tissue was used to compare measurements obtained from digital autoradiograms with classical gamma detector determination. This permits the evaluation of radioimager efficiency and absolute quantification. Radioimagers represent an improvement in terms of time of image acquisition. All the radioimagers are more sensitive than film for the detection of low levels of radioactivity. The spatial resolution provided by the mu-imager compares favourably with that obtained on film autoradiograms while digital autoradiograms from the phosphorimager and beta-imager did not show precise definition under our experimental conditions. Superimposition of histological structures from the stained sections with radiolabelled areas in the autoradiograms remains, at this time, the unique advantage of film. In conclusion, radioimagers represent an alternative to autoradiography on film or emulsion for in situ quantitative studies on tissue sections. They combine precise imaging for in situ binding studies with easy and direct access to counts in cpm. The improvement in radioimaging technology has, therefore, brought in situ analysis of iodinated ligand binding to the level of accuracy that is obtained with classical detectors of radioactivity.

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Imaging receptors by autoradiography: Computer-assisted approaches

Cited by 14 publications

References 18 publications

Receptor distribution in the human and animal hippocampus: Focus on muscarinic acetylcholine receptors

Receptor distribution in the human and animal hippocampus: Focus on muscarinic acetylcholine receptors

In vitro autoradiographic visualization of occupied estrogen receptors in the rat brain with an iodinated estrogen ligand.

Radioimagers as an alternative to film autoradiography forin situ quantitative analysis of125I-ligand receptor binding and pharmacological studies

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