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1.
Figure 4

Figure 4. From: Repeated swim impairs serotonin clearance via a corticosterone-sensitive mechanism: Organic cation transporter 3, the smoking gun.

Repeated swim reduces OCT3 expression and function in hippocampus. (A) Hippocampal OCT3 density was reduced in mice exposed to 14 d of swim, compared to no swim counterparts (percent normalized to 0 d controls; t-test, *p = 0.023, n = 10/group). Histamine clearance time (*P<0.05) (B) and rate (*P<0.01) (C) was slower in hippocampus of mice exposed to 14 d of swim.

NL Baganz, et al. J Neurosci. ;30(45):15185-15195.
2.
Figure 6

Figure 6. From: Repeated swim impairs serotonin clearance via a corticosterone-sensitive mechanism: Organic cation transporter 3, the smoking gun.

Adrenalectomy abolishes the swim-induced decrease in 5-HT clearance. (A) Maximal velocity (Vmax) for 5-HT clearance was slower in hippocampus of sham-operated mice exposed to 14 d of swim (2-way ANOVA with Bonferroni post-hoc test, ★p < 0.001), compared to sham-operated mice that were not exposed to swim and ADX mice swum for 0 or 14 d. (B) Plasma corticosterone concentration (ng/ml) was essentially undetectable in ADX mice, regardless of swim, compared to sham-operated mice (★P<0.01). Corticosterone levels were elevated in sham-operated mice exposed to 14 d of swim, compared to 0 d counterparts (**P<0.01).

NL Baganz, et al. J Neurosci. ;30(45):15185-15195.
3.
Figure 3

Figure 3. From: Repeated swim impairs serotonin clearance via a corticosterone-sensitive mechanism: Organic cation transporter 3, the smoking gun.

Serotonin clearance is impaired in SERT KO mice exposed to repeated swim. (A) As expected, in mice not exposed to swim, 5-HT clearance rate was slower in SERT KO (C57BL/6J background) mice than in C57BL/6J mice. Serotonin clearance rate in C57BL/6J mice from Figure 1 is re-plotted here for ease of comparison. In SERT KO mice, 5-HT clearance rate was slower in mice exposed to 14 d of swim, compared to unswum SERT KO mice (2-way ANOVA with Bonferroni post-hoc; *P<0.05; **P<0.01; ★P<0.01, n = 5–9/group/dose). (B) Summary of clearance rates for 20 pmol of 5-HT highlighting the dramatic effect of repeated swim to decrease the rate of 5-HT clearance in both C57BL/6J and SERT KO mice. (C) For a given amount of 5-HT (pmol) administered, 5-HT signal amplitudes (µM) were equivalent in all groups of mice. Serotonin signal amplitudes obtained in C57BL/6J mice from Figure 1 is re-plotted here for ease of comparison.

NL Baganz, et al. J Neurosci. ;30(45):15185-15195.
4.
Figure 5

Figure 5. From: Repeated swim impairs serotonin clearance via a corticosterone-sensitive mechanism: Organic cation transporter 3, the smoking gun.

The ability of corticosterone to inhibit 5-HT clearance in hippocampus is dependent on exposure to repeated swim. Change in 5-HT clearance rate (expressed as percent of baseline) 2 min post-hippocampal application of PBS control, EtOH (0.8 nmol), or corticosterone (272 pmol) in mice exposed to 0 or 14 d of swim. In both groups of mice, corticosterone inhibited 5-HT clearance rate, compared to PBS vehicle, but this effect of corticosterone was blunted in mice exposed to repeated swim (2-way ANOVA with Bonferroni post-hoc, *P<0.05; ★P<0.01). EtOH significantly reduced 5-HT clearance rate only in mice that were not exposed to swim. Pre-drug/vehicle signal amplitudes did not vary among groups. Pooled values were 3.59 ± 0.35 µM, n=17 and 3.20 ± 0.18 µM, n=18 for 0 d and 14 d swim respectively. Pre-drug/vehicle clearance rate was slower in mice exposed to 14 d swim (31.82 ± 5.56 nM/sec) compared to control mice (56.58 ± 8.4 nM/sec) (t33 = 2.595, p = 0.015).

NL Baganz, et al. J Neurosci. ;30(45):15185-15195.
5.
Figure 1

Figure 1. From: Repeated swim impairs serotonin clearance via a corticosterone-sensitive mechanism: Organic cation transporter 3, the smoking gun.

Serotonin clearance is impaired in the CA3 region of the hippocampus of mice subjected to repeated swim. (A) Representative oxidation currents (converted to 5-HT concentration, µM) produced by locally-applied 5-HT (barrel concentration 200 µM) in the CA3 region of the hippocampus of mice exposed to 0 d (black line) or 14 d (gray line) of swim. Raw traces are superimposed for ease of comparison. Time of clearance (T80) is defined by the amount of time it takes for the signal to decay to 80% of its peak amplitude (µM). Clearance rate (Tc) is the slope of the most linear portion of the descending signal ranging from T20 to T60. Compared to controls, 5-HT clearance rate (nM/sec) is slower (B) and time (sec) is longer (C) in hippocampus of mice swum for 14 d (2-way ANOVA with Bonferroni post-hoc comparison, *P<0.05; **P<0.01; ★P<0.01, n = 5–9/group/pmol amount). (D) For a given amount of 5-HT (pmol) administered, 5-HT signal amplitudes (µM) were equivalent in both groups of mice.

NL Baganz, et al. J Neurosci. ;30(45):15185-15195.
6.
Figure 2

Figure 2. From: Repeated swim impairs serotonin clearance via a corticosterone-sensitive mechanism: Organic cation transporter 3, the smoking gun.

SERT function and expression is the same in mice subjected to 0 or 14 d of repeated swim. As expected, fluvoxamine inhibited 5-HT clearance time (A) and rate (B) in hippocampus of control mice (no swim), however, these effects of fluvoxamine were equivalent in mice exposed to repeated swim. Compared to PBS vehicle, fluvoxamine significantly slowed 5-HT clearance time and rate in mice both groups of mice (2-way ANOVA with Bonferroni post-hoc, *P<0.05; **P<0.01, ★P<0.01, n = 5–9/group/dose). (C) In a behavioral read out of SERT and NET function, repeated swim did not affect the response to SERT and NET blockers in the tail suspension test. Immobility was sampled every 5 s over a 6 min test. Compared to saline, fluvoxamine (30 mg/kg), fluoxetine (15 mg/kg), and DMI (15 mg/kg) reduced immobility time in the TST in both groups of mice equally, regardless of swim. Data are expressed as a percent of respective saline control (2-way ANOVA with Bonferroni post-hoc; ★P<0.01).

NL Baganz, et al. J Neurosci. ;30(45):15185-15195.

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