AI Article Synopsis
- Fluorescent ligands, specifically BODIFY FL-prazosin (QAPB), allow for effective receptor study in tissues using confocal microscopy, offering advantages over traditional methods like antibodies.
- Histogram analysis of 3-D image data helps visualize and analyze large volumes of ligand-receptor binding under various conditions, including changes induced by antagonists.
- This method demonstrates sensitivity to receptor availability changes and can be applied beyond adrenergic receptors to other fluorescence-image-based assays.
Article Abstract
Fluorescent ligands provide the means of studying receptors in whole tissues using confocal laser scanning microscopy and have advantages over antibody- or non-fluorescence-based method. Confocal microscopy provides large volumes of images to be measured. Histogram analysis of 3-D image volumes is proposed as a method of graphically displaying large amounts of volumetric image data to be quickly analyzed and compared. The fluorescent ligand BODIFY FL-prazosin (QAPB) was used in mouse aorta. Histogram analysis reports the amount of ligand-receptor binding under different conditions and the technique is sensitive enough to detect changes in receptor availability after antagonist incubation or generic manipulations. QAPB binding was concentration dependent, causing concentration-related rightward shifts in histogram. In the presence of 10 microM phenoxybenzamine (blocking agent), the QAPB (50 nM) histogram overlaps the autofluorescence curve. The histogram obtained for the 1D knockout aorta lay to the left of that control and 1B knockout aorta, indicating a reduction in 1D receptors. We have shown, for the first time, that it is possible to graphically display binding of a fluorescent drug to a biological tissue. Although our application is specific to adrenergic receptors, the general method could be applied to any volumetric, fluorescence-image-based assay.
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| http://dx.doi.org/10.1162/15353500200504172 | DOI Listing |
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