Journal:

Article Title: Rapamycin Protects Against Neuron Death In in vitro and in vivo Models of Parkinson's Disease

doi: 10.1523/JNEUROSCI.3944-09.2010

Figure Lengend Snippet: (a) Mice were injected with either vehicle, rapamycin, MPTP or rapamycin + MPTP, as described in Methods for the acute regimen, and sacrificed after 24 hours. Ventral midbrains were dissected, homogenized in lysis buffer and the homogenates subjected to SDS-PAGE and Western immunoblotting. Membranes were probed with RTP801 antibody and reprobed with an antibody for total ERK1/2 as a loading control. Film resulting from the immunoblot was scanned and relative densities of RTP801 bands were normalized in each case to the densities of the corresponding ERK1/2 signals. Values (in arbitrary units) for RTP801 are expressed as mean ± SEM for at least three mice per condition. *p<0.01 vs mice injected with vehicle; ψp<0.01 vs mice injected with MPTP. (b) Mice subjected to the acute regimen as indicated were sacrificed 7 days after the last MPTP injection. Midbrains were cryo-sectioned, immunostained with tyrosine hydroxylase antibody and subjected to Nissl staining. Unbiased stereology was used to determine the total numbers of TH positive neurons in the substantia nigra. Results represent mean ± SEM of total number of TH-positive cells per substantia nigra (right and left) for at least 7 mice per each condition. **p<0.001 vs mice injected with vehicle; ++p<0.01 vs mice injected with rapamycin+MPTP. Lower panel shows representative micrographs of SN stained with TH-antibody in comparable sections for each condition. (c) Mice subjected to the subacute MPTP regimen as indicated were sacrificed two days after the last injection. Midbrains were cryo-sectioned, immunostained lightly with tyrosine hydroxylase antibody and subjected to Nissl staining to visualize condensed nuclei (with fragmented chromatin). Results represent mean ± SEM of total number of apoptotic nuclei per midbrain for at least 4 mice per each condition. ***p<0.001 vs mice injected with MPTP alone.

Article Snippet: In vivo MPTP treatments in mice Rapamycin preparation Rapamycin (LC laboratories, Woburn, MA) was dissolved in 0.2 ml of 100% ethanol and then diluted 100-fold with 40% propyleneglycol to obtain a final concentration of 0.75 mg/ml. ( Zhou et al., 2009 ) Animals and MPTP regimens Ten-week-old, male, C57/bl mice (Charles River Laboratories, Wilmington, MA) were divided into four groups of 3-10 mice (MPTP/rapamycin; MPTP/vehicle; saline/rapamycin; and saline/vehicle) and were subjected to either an acute or a sub-acute MPTP regimen.

Techniques: Injection, Lysis, SDS Page, Western Blot, Control, Staining