13 research outputs found
Dopamine D2 receptor pharmacology : In vitro analyses and in vivo pet imaging
The goals of the present thesis were to study high affinity agonist
binding at the D2 dopamine receptor and to explore the role of agonist
induced internalization of the receptor in positron emission tomography
(PET) imaging studies. For this purpose, we combined in vitro studies of
radioligand binding and receptor internalization with in vivo studies in
rodents using PET.
A pharmacological characterization of the novel PET radioligand MNPA in
vitro demonstrated that it is a potent and full agonist at the D2
receptor. In membrane homogenates, MNPA bound to both a high and low
affinity site of the D2 receptor, of which the high affinity site was
sensitive to guanosine triphosphate. In intact cells, however, MNPA bound
to only one site of low affinity. As a typical agonist at G-protein
coupled receptors, MNPA also induced receptor internalization, which was
further augmented by arrestin proteins.
[11C]MNPA was evaluated as an agonist radioligand and used to measure the
baseline occupancy of the D2 dopamine receptors by dopamine. In vivo,
[11C]MNPA was a D2 selective radioligand. Comparison of [11C]MNPA binding
in dopamine depleted rats and control rats suggested that about 50% of
the D2 receptors are occupied by dopamine at baseline.
To study high affinity agonist binding in vivo, we compared the binding
of [11C]MNPA in control mice to that of dopamine-beta-hydroxylase (DBH)
knockout mice, which have been reported to exhibit an increased
percentage of D2 receptors in the high affinity state. The in vivo
studies were accompanied by in vitro binding experiments in striatal
membrane homogenates. We did not observe any differences in [11C]MNPA
BPND between DBH knockout and control mice. Nor did we find any
differences in density or percentage of D2 receptors in high affinity
state with in vitro binding experiments. Combined, this study suggests
that DBH knockout mice have normal densities of D2 receptors in the high
affinity state.
Agonist induced D2 receptor internalization was evaluated using
immunohistochemistry in striatal tissue slices from arrestin3 knockout
and wild-type mice. Both dopamine and MNPA induced D2 receptor
internalization in wild-type tissue but not in knockout tissue. These
results demonstrated that D2 receptor internalization is mediated by
arrestin3 and that the arrestin3 knockout mice can be used to study D2
receptor internalization in vivo.
To determine whether the prolonged decrease of radioligand binding after
amphetamine is caused by receptor internalization, we imaged wild-type
and arrestin3 knockout mice, which are incapable of internalizing D2
receptors. The mice were imaged with both the D2 agonist [11C]MNPA and
the D2 antagonist [18F]fallypride. The effect of amphetamine on
radioligand binding was examined at two time points; 30 min and 4 hours
post-amphetamine. At 30 min, [11C]MNPA showed greater displacement than
[18F]fallypride, but each radioligand gave similar displacement in
knockout and wild-type mice. At 4 hours, the binding of both radioligands
returned to baseline in knockout mice but remained decreased in wild-type
mice. Our results suggest that the prolonged decrease of radioligand
binding after amphetamine is primarily due to D2 receptor internalization
rather than dopamine displacement.
In conclusion, the present thesis demonstrates that MNPA is a potent and
full agonist at the D2 dopamine receptor and binds in vitro to both a
high and low affinity state of the D2 receptor. We also demonstrated that
arrestin3 mediates internalization of the D2 dopamine receptor and that
the prolonged in vivo decrease of radioligand binding after amphetamine
is likely due to receptor internalization rather than dopamine
displacement
Visualization of Functional Neuropeptide Y Receptors in the Mouse Hippocampus and Neocortex Using [<sup>35</sup>S]GTPÎłS Binding
Kinetic modelling and test-retest reproducibility for the dopamine D\u2081R radioligand [\ub9\ub9C]SCH23390 in healthy and diseased mice
Synaptic vesicle glycoprotein 2A is affected in the CNS of Huntington\u2019s Disease mice and post-mortem human HD brain
Elevated type 1 metabotropic glutamate receptor availability in a mouse model of Huntington\u2019s disease : a longitudinal PET study
In vitro and In vivo assessment of suitable reference region and kinetic modelling for the mGluR1 radioligand <tex>\left[^{11}C\right]$</tex>ITDM in mice
Longitudinal preclinical evaluation of the novel radioligand [\ub9\ub9C]CHDI-626 for PET imaging of mutant huntingtin aggregates in Huntington's disease
PURPOSE: As several therapies aimed at lowering mutant huntingtin (mHTT) brain levels in Huntington’s disease (HD) are currently being investigated, noninvasive positron emission tomography (PET) imaging of mHTT could be utilized to directly evaluate therapeutic efficacy and monitor disease progression. Here we characterized and longitudinally assessed the novel radioligand [(11)C]CHDI-626 for mHTT PET imaging in the zQ175DN mouse model of HD. METHODS: After evaluating radiometabolites and radioligand kinetics, we conducted longitudinal dynamic PET imaging at 3, 6, 9, and 13 months of age (M) in wild-type (WT, n = 17) and heterozygous (HET, n = 23) zQ175DN mice. Statistical analysis was performed to evaluate temporal and genotypic differences. Cross-sectional cohorts at each longitudinal time point were included for post-mortem [(3)H]CHDI-626 autoradiography. RESULTS: Despite fast metabolism and kinetics, the radioligand was suitable for PET imaging of mHTT. Longitudinal quantification could discriminate between genotypes already at premanifest stage (3 M), showing an age-associated increase in signal in HET mice in parallel with mHTT aggregate load progression, as supported by the post-mortem [(3)H]CHDI-626 autoradiography. CONCLUSION: With clinical evaluation underway, [(11)C]CHDI-626 PET imaging appears to be a suitable preclinical candidate marker to monitor natural HD progression and for the evaluation of mHTT-lowering therapies. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00259-021-05578-8