Fluoxetine

Male SD rats were randomly divided into four groups: sham, sham+Fluoxetine, SAH+vehicle, and SAH+Fluoxetine group. Fluoxetine was purchased from Selleck and dissolved in sterile 0.9% NaCl. Fluoxetine (10 mg/kg) or vehicle was injected intravenously at 6 h after SAH induction as previously described [18 (link)]. SAH grade and neurological scores were measured in all groups. Brain edema (n = 6/group), Evans blue extravasation (n = 6/group), immunofluorescence/TUNEL staining (n = 5/group), quantitative real-time polymerase chain reaction (qRT-PCR) (n = 5/group), and western blot (n = 6/group) were performed.

Proliferation of cells was analyzed by Fluoroskan quantification following fluorescence labeling by stable transduction of the cell lines with a third generation lentiviral SIN vector carrying either the eGFP gene or the mCherry gene^{63 (link)}. Stimulation of cells was carried out in culture medium as detailed above containing indicated serum concentrations to ensure optimal steady-state conditions for cell growth. Labeled cells (1,000 cells/well for short-term (24–72 h) and 500 cells/well for long-term (96–144 h)) were incubated in flat bottom 96-well plates in the absence (controls) or in the presence of the SSRIs fluoxetine, sertraline, and citalopram (Selleckchem) at clinically relevant concentrations (10–1000 nM) as well as with 5-HT (Selleckchem). At indicated time points, the medium was removed and cells were lysed with 10% SDS following detection of fluorescence intensity of mCherry (excitation 485 nm, emission 612 nm) and eGFP (excitation 485 nm, emission 520 nm) using the Fluoroskan Ascent FL (Thermo Fisher Scientific, Schwerte, Germany) since the fluorescence intensity is proportional to the cell number^{59 (link)}. Relative proliferation rates were calculated as percentage of controls.