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. Author manuscript; available in PMC: 2021 Jun 29.
Published in final edited form as: Cell Rep. 2021 May 11;35(6):109123. doi: 10.1016/j.celrep.2021.109123

Figure 2. DAT expression and function in DAT-P2A-Flpo mice.

Figure 2.

(A) Representative western blot images of DAT, TH, vesicular monoamine transporter 2 (VMAT2), and histone H3 (H3) loading control from striatal lysates from DAT-P2A-Flpo WT and heterozygous (Het) mice. Two independent samples per genotype are shown. Blots were cropped to show the relevant bands. Molecular weight (MW) in kDa is indicated on the right. Representative of 4 mice.(B) Quantification of protein levels relative to H3, normalized to WT. Bars represent mean ± SEM. Each dot represents the average of two striatal samples from one mouse (n = 4 mice per genotype, 2 males and 2 females, P100–120). DAT, *p = 0.0259; TH, p = 0.9639; VMAT2, p = 0.0930; unpaired t tests.

(C) Extracellular DA ([DA]o) transients evoked by single electrical pulses and recorded with fast-scan cyclic voltammetry (FCV) in different striatal sub-regions. Traces are mean ± SEM [DA]o versus time (average of 32 transients per recording site from 4 pairs of DAT-P2A-Flpo WT and Het mice, 2 male pairs and 2 female pairs, P100–120). 1, ventromedial striatum; 2, dorsomedial striatum; 3, dorsolateral striatum; 4, central striatum; 5, ventrolateral striatum; 6+7, nucleus accumbens (NAc).

(D) Mean ± SEM [DA]o versus time (average of 80 transients per genotype) from the dorsal striatum (dStr) sites 1–5.

(E) Mean ± SEM [DA]o versus time (average of 32 transients per genotype) from the NAc sites 6+7.

(F) Mean ± SEM [DA]o of selected region- and peak-matched transients from the dStr of DAT-P2A-Flpo WT and Het mice (average of 22 transients per genotype).

(G) Single exponential curve fit of the decay phase of the transients in (F). DAT-P2A-Flpo Het mice have significantly slower [DA]o re-uptake than WT mice (***p < 0.0001; WT tau = 0.409, Het tau = 0.648).

(H–K) Behavioral performance of DAT-P2A-Flpo mice in a 60-min open-field test. For all graphs, bars represent mean ± SEM and dots represent values for individual mice. n = 15 WT mice, 6 males and 9 females; n = 14 Het mice, 5 males and 9 females, P50–90. (H) Total distance traveled in 60 min; p = 0.2680, unpaired t test. (I) Total number of rears in 60 min; p = 0.8847, unpaired t test. (J) Total time spent in the center of open field in 60 min; p = 0.9981, unpaired t test. (K) Total number of grooming bouts in the first 20 min; p = 0.8061, unpaired t test.

See also Figure S2.