TY - JOUR
T1 - Computer-assisted video analysis of [3H]spiroperidol binding autoradiographs
AU - Anthony Altar, C.
AU - Walter, Robert J.
AU - Neve, Kim A.
AU - Marshall, John F.
N1 - Funding Information:
The visual image processing equipment is part of the Laser Microbeam Program (LAMP) established in the Department of Developmental and Cell Biology at the University of California, Irvine, and is part of the National Institutes of Health Biotechnology Resource Program of the Division of Research Resources. This work has been supported in part by NIH Grants HL 15740, GM 23445, and RRO 1192 awarded to M.W. Berns and NSF Grant BNS 8208656 awarded to J.F.M.C.A.A. was supported by NRSA Postdoctoral Training Grant AG 00096 from the NIA. K.A.N. was supported by NRSA Predoctoral Fellowship MH 08915 from the NIMH. The assistance of Dr. Herbert P. Killackey in identifying neocortical distributions of binding is greatly appreciated. We thank Mary King for typing the manuscript.
PY - 1984/3
Y1 - 1984/3
N2 - A computer-based video image processing system is described that quantifies the binding of the neuroleptic drug, [3H]spiroperidol, to rat forebrain sections. Adjacent sections were incubated in buffer containing [3H]spiroperidol or [3H]spiroperidol plus 1 μM (+)-butaclamol and exposed to tritium-sensitive film to produce autoradiographs of total binding or non-butaclamol-displaced ('non-specific') binding. The image processing system digitized each autoradiograph, attenuated geometric distortion and unevenness of background illumination, and reassigned the digitized image intensities (gray values) to be a linear function of fmol [3H]spiroperidol bound per mg protein by using a calibration curve generated from 3H-containing tissue standards. An image of butaclamol-displaced ('specific') [3H]spiroperidol binding was produced by subtracting the linearized image of non-specific binding from the superimposed image of total binding. An image of percent specific [3H]spiroperidol binding was obtained by dividing the image of specific binding by the superimposed image of total binding. The computer-derived images, which could be displayed in gray tones or pseudocolor-coded, revealed that the greatest amounts of specific [3H]spiroperidol binding (1000-3000 fmol/mg protein: 60-80% specifically bound) were located in layers 5A and 5C of the neocortex, the claustrum, and in the caudate-putamen, where a lateral-to-medial binding gradient occurred. [3H]Spiroperidol was bound to a lesser extent (400-1000 fmol/mg protein; 31-51% specifically bound) to the medial nucleus accumbens septi or olfactory tubercle and without measurable specificity to the lateral septum, anterior commissure, corpus callosum, or superficial neocortex. These procedures are particularly useful for the quantitative and visual analysis of autoradiographs in which [3H]ligand binding is associated with more than a single site.
AB - A computer-based video image processing system is described that quantifies the binding of the neuroleptic drug, [3H]spiroperidol, to rat forebrain sections. Adjacent sections were incubated in buffer containing [3H]spiroperidol or [3H]spiroperidol plus 1 μM (+)-butaclamol and exposed to tritium-sensitive film to produce autoradiographs of total binding or non-butaclamol-displaced ('non-specific') binding. The image processing system digitized each autoradiograph, attenuated geometric distortion and unevenness of background illumination, and reassigned the digitized image intensities (gray values) to be a linear function of fmol [3H]spiroperidol bound per mg protein by using a calibration curve generated from 3H-containing tissue standards. An image of butaclamol-displaced ('specific') [3H]spiroperidol binding was produced by subtracting the linearized image of non-specific binding from the superimposed image of total binding. An image of percent specific [3H]spiroperidol binding was obtained by dividing the image of specific binding by the superimposed image of total binding. The computer-derived images, which could be displayed in gray tones or pseudocolor-coded, revealed that the greatest amounts of specific [3H]spiroperidol binding (1000-3000 fmol/mg protein: 60-80% specifically bound) were located in layers 5A and 5C of the neocortex, the claustrum, and in the caudate-putamen, where a lateral-to-medial binding gradient occurred. [3H]Spiroperidol was bound to a lesser extent (400-1000 fmol/mg protein; 31-51% specifically bound) to the medial nucleus accumbens septi or olfactory tubercle and without measurable specificity to the lateral septum, anterior commissure, corpus callosum, or superficial neocortex. These procedures are particularly useful for the quantitative and visual analysis of autoradiographs in which [3H]ligand binding is associated with more than a single site.
KW - autoradiography spiroperidol
KW - basal ganglia
KW - computer-assisted image processing
KW - neostriatum
KW - receptor binding
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U2 - 10.1016/0165-0270(84)90054-2
DO - 10.1016/0165-0270(84)90054-2
M3 - Article
C2 - 6738107
AN - SCOPUS:0021264303
SN - 0165-0270
VL - 10
SP - 173
EP - 188
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
IS - 3
ER -