Quantitative Autoradiography as a Tool to Study Receptors in Neural Tissue. Studies on 3H-Ouabain Binding Sites and Correlation with Synaptic Protein Phosphorylation in Different Brain Areas

Localization and Characterization of Binding Sites with High Affinity for [3H]Ouabain in Cerebral Cortex of Rabbit Brain Using Quantitative Autoradiography

“…Recently, several investigators (Spyropoulos and Rainbow, 1984; Benfenati et al, 1985;Caspera et al, 1987) have carried out QAR with rat brain tissue sections labeled with radioactive cardiac glycosider;; however, little has been reported on the specificity, kinetics, or characteristics of binding. The aims of the present study were to localize specific ouabain binding sites in cerebral cortex and to determine if the characteristics of these sites, measured by QAR, are the same as those determined in biochemical studies with isolated microsomal membranes.…”

Quantitative autoradiography of central neurotransmitter receptors: methodological and statistical aspects with special reference to computer‐assisted image analysis

“…Another interesting comparison made to verify the reliability of autoradiographical determinations with respect to biochemical studies was carried out at the spinal cord and neocortical level. In a previous autoradiographical study (Benfenati et al 1985) it was reported that [3H]ouabain binding is particularly high in the spinal cord grey matter and rather low in the fronto-parietal cortex. However, subsequent determinations carried out on crude membrane preparations or mitochondrial (P2) fractions from neocortex and spinal cord did not show the previously observed difference.…”

“…Receptor autoradiography was performed on 14-,um cryostat sections obtained from the brain of Sprague-Dawley rats (body weight Z O W Z~O g) kept under standardized dark and light conditions; the rats were given food pellets and water ad libitum. The procedure for tissue preparation and sectioning has been extensively described elsewhere (Herkenham & Pert 1982, Fuxe et al 1983, Agnati et al 1984c, Benfenati et al 1985. Briefly rats, under a light ketamine anaesthesia, were perfused via an aortic cannula with 50 ml saline solution followed by 300 ml ice-cold phosphatebuffered saline containing 0.…”

“…I yo paraformaldehyde at pH 7.4 for 20-30 min; brains were removed, frozen with CO, and cut at -18 "C. Particular care was used in melting and drying sections on gelatinized slides at o "C under humidity-free atmosphere (Herkenham 8z Pert 1982). Sections were then incubated under appropriate conditions (Schwann et al 1982, Agnati et al 1984c, Benfenati et al 1985 with [3H]spiperone (25.0 Ci mmol-I, New England Nuclear, Boston, Mass., USA) and [3H]ouabain (20.8 Ci mmol-I, New England Nuclear) in order to visualize dopamine and Na-K-ATPase binding sites, respectively. In order to compare absolute binding measurements performed on tissue sections with biochemical determinations, [3H]ouabain binding was also assayed biochemically in cellular subfractions prepared from cerebral cortex and spinal cord (De Robertis et a f .…”

“…On the occasion of the 20th Anniversary of the Stockholm 'Amine Group', November 1985. , Unnerstall et al 1982, Rainbow et al 1982, Benfenati et al 1985 in order to study the distribution of neurotransmitter receptors in the brain (Unnerstall et al. , Fuxe et al 1983), their biochemical characteristics and functional changes in different experimental conditions (Palacios 1984, Agnati et al 1 9 8 4~) .…”