Ibutamoren Mesylate

Animal experiment:

Compounds used are: Ibutamoren mesylate (50 μg), octreotide (100 μg), and mGH (30 μg). Mice are give an initial ip injection (0.1 mL) of either saline, octreotide or mGH, followed 10 min later by an ip injection (0.1 mL) of either saline or Ibutamoren mesylate. Thus, the first study comprised of the following groups: saline/saline, saline/Ibutamoren mesylate, mGH/saline, mGH/Ibutamoren mesylate saline/saline, saline/Ibutamoren mesylate, mGH/saline, mGH/Ibutamoren mesylate, and the second study of: saline/saline, saline/Ibutamoren mesylate, octreotide/saline, octreotide/Ibutamoren mesylate. Additionally, a number of mice are injected ip with hypertonic saline (0.2 mL, 1.5 M) to serve as positive controls for the immunocytochemistry. Ninety minutes after injection animals are terminally anesthetized with sodium pentobarbitone (60 mg/kg, ip) and perfused transcardially with heparinized saline followed by 4% paraformaldehyde in 0.1mol/Lphosphate buffer (PB, pH 7.4). Brains are then removed and placed in the same solution for 24 h before being stored at− 80°C until processing. Coronal sections of forebrain (40 μM) are cut on a freezing microtome and placed in 0.1mol/LPB containing Triton X-100 (PB-T, pH 7.4). Sections are incubated for 24 h at 4°C in Ab-2 Fos antibody (rabbit polyclonal; 1:1000 in 1% normal sheep serum). The antibody-antigen complex is localized with a 1-h incubation in biotinylated anti-rabbit Ig, followed by a 1-h incubation in avidin, biotinylated horseradish peroxidase. The reaction product is visualized using a glucose oxidase-diaminobenzidine-nickel method, and Fos-like immunoreactivity is visualized as a dense purple-black precipitate restricted to the nucleus. The number of c-fos positive nuclei in the arcuate and periventricular nuclei (six to eight sections per mouse) are counted double-blind and a group mean calculated (mean±sem). Statistical analysis is performed by a two-way ANOVA followed by an all pairwise multiple comparison of data with significance taken as P < 0.05.

References:

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