Rotenone - Molecule of the Month January 2004 [Archived version]

This is the Molecule of the Month entry for January 2004 about rotenone. It is a pdf archive version of the HTML webpage

Immunostaining for activated caspase 3 to demonstrate apoptosis.

<p>An acute dose of GDNF (0.1, 0.2, 0.6 and 1.0 µg/µL) or aCSF vehicle control was infused into the striatum and tissue was collected after 2 weeks and 4 weeks. Representative images showing cells immuno-positive for activated caspase 3 (red) and NeuN (green). Scale bar = 100 µm.</p

Number of hemispheres infused with r-metHuGDNF for qPCR, ELISA, and immunohistochemical analysis.

<p>Number of hemispheres infused with r-metHuGDNF for qPCR, ELISA, and immunohistochemical analysis.</p

Immunostaining for NeuN to show neuronal cell loss.

<p>Neuonal cell loss was seen around the r-metHuGDNF infused area after 2 weeks and 4 weeks at 0.6 µg/µL and 1.0 µg/µL. A lack of NeuN (red) was generally visible where GDNF (green) had been infused at these concentrations (see inlay). Nuclei are marked in blue with the use of DAPI. Scale bar = 250 µm.</p

Pharmokinetics of r-metHuGDNF.

<p>(A) ELISA to show amount of total human GDNF protein (pg/mg of total protein) within the striatum after 2 weeks and 4 weeks following acute CED of 0.1, 0.2, 0.6, and 1.0 µg/µL r-metHuGDNF into the striatum. The flow rate employed was 1 µL/minute. Each bar indicates the mean ± SD. **<i>p</i><0.01, r-metHuGDNF infused hemispheres versus aCSF control infused hemispheres by one-way ANOVA and Dunnett post-hoc test. (B) Representative images of immunostaining for GDNF after 2 weeks and 4 weeks following acute CED of 0.1, 0.2, 0.6, and 1.0 µg/µL r-metHuGDNF into the striatum. Scale bar = 250 µm.</p

Expression of GAP43 mRNA and synaptophysin.

<p>Levels of GAP43 mRNA were analysed following Q-PCR analysis (A). Synaptophysin protein levels were measured using Sandwich ELISA (B). Each bar indicates the mean ± SD. **<i>p</i><0.01, r-metHuGDNF infused hemispheres versus aCSF control infused hemispheres.</p

Immunostaining for Glial Fibrillary Acidic Protein (GFAP).

<p>GFAP-immunopositive cells were observed in the tissue surrounding the needle track area at both 2 and 4 weeks (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056186#pone-0056186-g003" target="_blank">Figure 3</a>). There was no detectable correlation between the degree of gliosis adjacent to the needle track and the infused concentration of r-metHuGDNF at any time point. Scale bar = 500 µm.</p

Immunostaining for Iba1, a protein upregulated during the activation of microglia.

<p>Immunostaining is shown 2 weeks and 4 weeks after r-metHuGDNF infusion. Iba1 (red) was seen in all infused concentrations at 2 weeks. The inlay shows the presence of perivascular microglia. GDNF is shown in green and nuclei marker, DAPI, in blue. Scale bar = 250 µm.</p