SPF mice were orally gavaged daily for 7 days with 10 mg/kg fluoxetine (Santa Cruz) or treated with fluoxetine (40 μg/ml) in the drinking water for 14 days as indicated in figure legends. Fecal samples were harvested on day 0, 1, 4, 7 or 14 of treatment as indicated in figure legends and processed for 16S rDNA sequencing as described above.
Fluoxetine
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Fluoxetine is a laboratory reagent used in research applications. It is a selective serotonin reuptake inhibitor (SSRI) that is commonly used as a reference compound in various experimental studies.
Automatically generated - may contain errors
Lab products found in correlation
Vacuum manifold, by Merck Group (2 mentions)
Ls6500, by Beckman Coulter (2 mentions)
Reserpine, by Merck Group (2 mentions)
Tetrabenazine, by Merck Group (2 mentions)
Unlabeled 5 ht, by Merck Group (2 mentions)
Ne or trp, by PerkinElmer (2 mentions)
Multiscreenhts plates, by Merck Group (2 mentions)
Filter count scintillation fluid, by PerkinElmer (2 mentions)
6 protocols using fluoxetine
1
Fluoxetine Modulates Gut Microbiome
2
Fluoxetine Modulates Gut Microbiome
3
Microbial Uptake of Neurotransmitters
4
Microbial Uptake of Neurotransmitters
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T. sanguinis or B. theta was cultured anaerobically as described above and subcultured for 15–17 hours to reach stationary phase. Cells were washed and resuspended in Kreb-Ringer’s buffer, then left untreated or pre-treated with vehicle, unlabeled 5-HT (Sigma Aldrich), fluoxetine (Santa Cruz), reserpine (Sigma Aldrich) or tetrabenazine (Sigma Aldrich) for 30 min at 37°C. Cells were then incubated with 1 uM tritiated 5-HT (plus 0–500 uM unlabeled 5-HT for dose-dependent assays), NE or Trp (Perkin Elmer). For T. sanguinis, all uptake reactions were performed at room temperature. For B. theta, uptake reactions were performed at either room temperature or 37°C.
Uptake reactions were terminated by washing in ice cold Kreb-Ringer’s buffer. Washed T. sanguinis was trapped through 0.45 um PVDF filters on MultiScreenHTS plates connected to a vacuum manifold (EMD Millipore). For B. theta, washed cells were either trapped through 0.45 um PVDF filters or lysed by incubating in TE buffer plus triton and lysozyme. Extracted filters or lysed cells were incubated in 4 ml Filter Count scintillation fluid (Perkin Elmer) for 1 hour at room temp and radioactivity was measured using a liquid scintillation counter (Beckman LS6500).
Uptake reactions were terminated by washing in ice cold Kreb-Ringer’s buffer. Washed T. sanguinis was trapped through 0.45 um PVDF filters on MultiScreenHTS plates connected to a vacuum manifold (EMD Millipore). For B. theta, washed cells were either trapped through 0.45 um PVDF filters or lysed by incubating in TE buffer plus triton and lysozyme. Extracted filters or lysed cells were incubated in 4 ml Filter Count scintillation fluid (Perkin Elmer) for 1 hour at room temp and radioactivity was measured using a liquid scintillation counter (Beckman LS6500).
5
Chronic SSRI Administration in Mice
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Following initial PCA/saline injections and a 2 d recovery period, mice began daily injections of SSRI or saline intraperitoneally (Fig. 1C ). Fluoxetine (Santa Cruz, catalog #sc-201125A) was dissolved in saline at concentrations of 2 mg/ml and 1 mg/ml and administered to mice at 20 mg/kg or 5 mg/kg, respectively. Sertraline (Acros Organics, catalog #462190010) was dissolved in 1.5% DMSO in saline at a concentration of 1 mg/ml and administered to mice at 10 mg/kg. Saline controls for sertraline animals received 1.5% DMSO in saline. Once daily injections were given for either 6 weeks or 10 weeks depending upon the experiment.
6
In Vitro and In Vivo Pharmacological Testing
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For the in vitro study, the compounds (TKDC and 28NH) were dissolved in dimethyl sulfoxide. For the in vivo study, TKDC was dissolved in 5‰ tween-80 and intraperitoneally administered to animals. Used as a positive control, fluoxetine (Santa Cruz Biotechnology, USA) was dissolved in water intraperitoneally administered at a dose of 10 mg kg−1. The vehicle, 5‰ tween-80, was administered to control mice as described above. All animals were treated with the drugs 30 min before the behavioral experiments.
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