High-resolution phosphor imaging: validation for use with human brain tissue sections to determine the affinity and density of radioligand binding.
This study investigated the suitability of high-resolution storage phosphor imaging for the quantitative analysis of radioligand binding to human brain tissue. Hence, the binding of [(3)H]mazindol to the dopamine transporter in caudate-putamen tissue homogenates or frozen tissue sections apposed to either autoradiographic film or phosphor imaging plates was measured. Estimates of binding affinity were similar for homogenate studies and phosphor imaging plates (Kd=6.44+/-0.14 and 6.91+/-0.47 nM, respectively), but higher values were obtained with film autoradiography (Kd=11.31+/-0.82 nM). The density of binding was similar for both autoradiographic techniques (Bmax=371.9+/-30.8 fmol/mg estimated tissue equivalent, ETE (imaging plate) and 425+/-13.77 fmol/mg ETE (film)), although lower values were obtained from tissue homogenates (Bmax=64.27+/-6.74 fmol/mg wet weight). These results suggest that high resolution phosphor imaging can be used to analyse radioligand binding parameters in human brain tissue. Moreover, the reduced exposure time of phosphor imaging plates (e.g. 7 days vs 5 weeks) allows results to be obtained more rapidly than with conventional film autoradiography.
Pavey
Geoffrey M
GM
The Rebecca L. Cooper Research Laboratories, Division of Molecular Schizophrenia, The Mental Health Research Institute of Victoria, Locked Bag 11, Parkville, Vic. 3052, Australia. g.pavey@papyrus.mhri.edu.au
Copolov
David L
DL
Dean
Brian
B
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001345:Autoradiography / Q000295:instrumentation / Q000379:methods; D001921:Brain / Q000378:metabolism*; D002421:Caudate Nucleus / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System / Q000378:metabolism; D050483:Dopamine Plasma Membrane Transport Proteins; D018765:Dopamine Uptake Inhibitors / Q000378:metabolism; D005260:Female; D006801:Humans; D007089:Image Enhancement / Q000379:methods; D008297:Male; D008454:Mazindol / Q000378:metabolism; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins / Q000378:metabolism; D008875:Middle Aged; D009419:Nerve Tissue Proteins; D011699:Putamen / Q000378:metabolism; D011869:Radioligand Assay / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data
D050483:Dopamine Plasma Membrane Transport Proteins; D018765:Dopamine Uptake Inhibitors; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D008454:Mazindol