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. 2015 Dec 16;35(50):16463–16478. doi: 10.1523/JNEUROSCI.2131-15.2015

Figure 4.

Figure 4.

Adoptive transfer of splenocytes from mice treated with VIPR agonists is neuroprotective. A, Photomicrographs of TH+Nissl+ neurons in the SN and STR in mice treated with PBS, MPTP, or MPTP followed by adoptive transfer of spleen cells from mice treated with VIP, LBT-3393, or LBT-3627 (40× image; scale bar, 200 μm). Sections were immunostained with anti-TH and HRP-conjugated secondary antibody and visualized with DAB. SN sections were counterstained with thionin. B, Representative photomicrographs of Mac-1+ microglia within the SN of mice treated with PBS alone, MPTP alone, or MPTP-treated mice receiving splenocytes from donors treated with VIP, LBT-3393, or LBT-3627 (40× image; scale bar, 200 μm; inset = 200× image). C, Total numbers of surviving dopaminergic neurons (TH+Nissl+) and nondopaminergic neurons (TH−Nissl+) in the SN after MPTP treatment and adoptive transfer (10× image; scale bar, 1000 μm). D, Relative TH densitometry of striatal dopaminergic termini. E, Quantification of reactive microglia taken from midbrains 2 d after MPTP treatment. Sections were stained for Mac-1+ microglia using an anti-Mac-1 antibody, HRP-conjugated secondary antibody, and DAB for color visualization. Numbers of reactive microglia (amoeboid Mac-1+) were determined by stereological analysis. C, D, Differences in means (±SEM, n = 8) were determined where p < 0.05 compared with groups treated with PBS (a), MPTP (b), VIP (c), or LBT-3393 (d). E, Differences in means (±SEM, n = 5) were determined where p < 0.05 compared with groups treated with PBS (a) or MPTP alone (b).