Receptor imaging technique with 11C-labeled receptor ligands in living brain slices: Its application to time-resolved imaging and saturation analysis of benzodiazepine receptor using [11C]Ro15-1788

Tetsuhito Murata, Kiyoshi Matsumura, Hirotaka Onoe, Mats Bergström, Hajime Takechi, Sven Sihver, Wiebke Sihver, Henrik Neu, Yvonne Andersson, Mattias Ögren, Karl Johan Fasth, Bengt Långström, Yasuyoshi Watanabe

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Abstract

Recently we developed a novel imaging technique using positron emitter-labeled compounds as probes and a storage phosphor screen as a detector. This approach makes it possible to follow a variety of biochemical processes with spatial information in living brain slices. Further technical development is reported here in terms of time-resolved imaging and receptor characterization in a real equilibrium state. The method was validated by use of [11C]Ro15-1788, a benzodiazepine receptor antagonist. Fresh brain slices were incubated with [11C]Ro15-1788 in oxygenated Krebs-Ringer solution at 37°C, in a specially designed chamber. By placing the chamber on a storage phosphor screen, we could obtain two-dimensional images of radioactivity in the slices. Time-resolved imaging was made at 5 min intervals, revealing that it took 60 min to reach equilibrium binding. The dissociation process was observed by adding an excess amount of unlabeled Ro15-1788 to the chamber, 25 min was required for the full dissociation. In the equilibrium state, i.e. in the presence of free radio-ligand, Scatchard plot analysis was performed on the cerebral cortex (K(d) = 7.4 nM, B(max) = 146 fmol/mg tissue) and striatum (K(d) = 7.5 nM, B(max) = 107 fmol/mg tissue), suggesting the presence of a single component of binding site in these two regions. The present method, for the first time, made it possible to study a ligand-receptor interaction in living brain slices with temporal and spatial resolutions. This technique should prove useful for studies of receptor function under physiological conditions.

Original languageEnglish
Pages (from-to)145-154
Number of pages10
JournalNeuroscience Research
Volume25
Issue number2
DOIs
Publication statusPublished - 01-01-1996

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GABA-A Receptors
Ligands
Brain
Biochemical Phenomena
Radio
Cerebral Cortex
Radioactivity
Binding Sites
Electrons

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

Murata, Tetsuhito ; Matsumura, Kiyoshi ; Onoe, Hirotaka ; Bergström, Mats ; Takechi, Hajime ; Sihver, Sven ; Sihver, Wiebke ; Neu, Henrik ; Andersson, Yvonne ; Ögren, Mattias ; Fasth, Karl Johan ; Långström, Bengt ; Watanabe, Yasuyoshi. / Receptor imaging technique with 11C-labeled receptor ligands in living brain slices : Its application to time-resolved imaging and saturation analysis of benzodiazepine receptor using [11C]Ro15-1788. In: Neuroscience Research. 1996 ; Vol. 25, No. 2. pp. 145-154.
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abstract = "Recently we developed a novel imaging technique using positron emitter-labeled compounds as probes and a storage phosphor screen as a detector. This approach makes it possible to follow a variety of biochemical processes with spatial information in living brain slices. Further technical development is reported here in terms of time-resolved imaging and receptor characterization in a real equilibrium state. The method was validated by use of [11C]Ro15-1788, a benzodiazepine receptor antagonist. Fresh brain slices were incubated with [11C]Ro15-1788 in oxygenated Krebs-Ringer solution at 37°C, in a specially designed chamber. By placing the chamber on a storage phosphor screen, we could obtain two-dimensional images of radioactivity in the slices. Time-resolved imaging was made at 5 min intervals, revealing that it took 60 min to reach equilibrium binding. The dissociation process was observed by adding an excess amount of unlabeled Ro15-1788 to the chamber, 25 min was required for the full dissociation. In the equilibrium state, i.e. in the presence of free radio-ligand, Scatchard plot analysis was performed on the cerebral cortex (K(d) = 7.4 nM, B(max) = 146 fmol/mg tissue) and striatum (K(d) = 7.5 nM, B(max) = 107 fmol/mg tissue), suggesting the presence of a single component of binding site in these two regions. The present method, for the first time, made it possible to study a ligand-receptor interaction in living brain slices with temporal and spatial resolutions. This technique should prove useful for studies of receptor function under physiological conditions.",
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Murata, T, Matsumura, K, Onoe, H, Bergström, M, Takechi, H, Sihver, S, Sihver, W, Neu, H, Andersson, Y, Ögren, M, Fasth, KJ, Långström, B & Watanabe, Y 1996, 'Receptor imaging technique with 11C-labeled receptor ligands in living brain slices: Its application to time-resolved imaging and saturation analysis of benzodiazepine receptor using [11C]Ro15-1788', Neuroscience Research, vol. 25, no. 2, pp. 145-154. https://doi.org/10.1016/S0168-0102(96)01037-1

Receptor imaging technique with 11C-labeled receptor ligands in living brain slices : Its application to time-resolved imaging and saturation analysis of benzodiazepine receptor using [11C]Ro15-1788. / Murata, Tetsuhito; Matsumura, Kiyoshi; Onoe, Hirotaka; Bergström, Mats; Takechi, Hajime; Sihver, Sven; Sihver, Wiebke; Neu, Henrik; Andersson, Yvonne; Ögren, Mattias; Fasth, Karl Johan; Långström, Bengt; Watanabe, Yasuyoshi.

In: Neuroscience Research, Vol. 25, No. 2, 01.01.1996, p. 145-154.

Research output: Contribution to journalArticle

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T1 - Receptor imaging technique with 11C-labeled receptor ligands in living brain slices

T2 - Its application to time-resolved imaging and saturation analysis of benzodiazepine receptor using [11C]Ro15-1788

AU - Murata, Tetsuhito

AU - Matsumura, Kiyoshi

AU - Onoe, Hirotaka

AU - Bergström, Mats

AU - Takechi, Hajime

AU - Sihver, Sven

AU - Sihver, Wiebke

AU - Neu, Henrik

AU - Andersson, Yvonne

AU - Ögren, Mattias

AU - Fasth, Karl Johan

AU - Långström, Bengt

AU - Watanabe, Yasuyoshi

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