WT and MPS-IIIA mice were used to test the effect of α-methyl-para-tyrosine (α-MPT) on autistic-like behaviours. α-MPT (250 mg/kg i.p.; Sigma Aldrich) was dissolved in NaCl 0.9%. It was injected 2 h before starting behavioural tests. Mice were tested first in the social dominance tube test that was immediately followed by the open field test on the same day to avoid the re-injection of α-MPT on the following days (related to Fig. 3a, a ʺ). We selected the social tube test because it has a lower mean duration with respect to the social novelty task, in this way avoiding a longer delay post injection. Another group of mice was used to test the effect of α-MPT in the open field test 5 h after the injection (later time-point) (related to Supplementary Fig. 3g ). The dose of the drug and the testing time were chosen based on literature defining the time- and dose-dependent biochemical and behavioural effects of α-MPT32 ,60 ,61 . After the behavioural tests, the mice were sacrificed by cervical dislocation. They were not used for any biochemical or histological experiment, due to the effects of α-MPT on DA depletion.
α mpt
Manufactured by Merck Group
Sourced in France
α-MPT is a laboratory reagent used in research applications. It functions as an α-ketoglutarate dehydrogenase inhibitor.
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2 protocols using α mpt
1
Evaluating Effect of α-MPT on Autism-like Behaviors
2
Measuring Catecholamine Turnover Dynamics
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Catecholamine turnover was measured on the basis of the decline in tissue NE content after the inhibition of catecholamine biosynthesis with α-methyl-dl -tyrosine (α-MPT) (200 mg/kg i.p. injection; Sigma-Aldrich, ST, Quentin, France), as described previously (44 (link)).
In the morning, bedding was changed, and C57BL/6N mice were food-deprived for 3 hours to insure postprandial state and injected with α-methyl-dl -tyrosine (α-MPT; a tyrosine hydroxylase inhibitor) to block catecholamine synthesis. Before (time = 0) and 3 hours after the injection (time = 3 hours), animals were euthanized, and the tissues were removed, flash-frozen in liquid nitrogen, and stored at −80°C for monoamine and metabolite analysis.
Catecholamine content at time = 0 [NE (0)] was determined on a group of animals receiving a saline injection. Because the concentration of catecholamine in tissues declined exponentially, we could obtain the rate constant of NE efflux (expressed in h − 1). Comprehensive analysis of NE was carried out by reverse-phase liquid chromatography (LC) with electrochemical detection as described in (67 ). The values obtained were expressed as nanogram per milligram wet tissue and were logarithmically transformed for calculation of linearity of regression, SE of the regression coefficients, and significance of differences between regression coefficients.
In the morning, bedding was changed, and C57BL/6N mice were food-deprived for 3 hours to insure postprandial state and injected with α-methyl-
Catecholamine content at time = 0 [NE (0)] was determined on a group of animals receiving a saline injection. Because the concentration of catecholamine in tissues declined exponentially, we could obtain the rate constant of NE efflux (expressed in h − 1). Comprehensive analysis of NE was carried out by reverse-phase liquid chromatography (LC) with electrochemical detection as described in (67 ). The values obtained were expressed as nanogram per milligram wet tissue and were logarithmically transformed for calculation of linearity of regression, SE of the regression coefficients, and significance of differences between regression coefficients.
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