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. Author manuscript; available in PMC: 2018 Mar 1.
Published in final edited form as: Acta Neuropathol. 2016 Dec 23;133(3):381–393. doi: 10.1007/s00401-016-1661-y

Fig. 5.

Fig. 5

Markers of transduction in the medulla oblongata and midbrain of AAV-injected rats. a Rats (n = 6) received midbrain injections of hα-synuclein-AAVs and were killed after 2 months. Non-injected (NI) animals provided control samples. Hα-synuclein (hα-syn), WPRE or hypoxantine phosphoribosyltransferase 1 (HPRT) mRNA was assayed by RT-PCR in samples of the right (AAV-injected) ventral mesencephalon (VM) or dorso-medial medulla oblongata (MO). Specific bands were at 79 (hα-syn), 204 (WPRE), and 61 (HPRT) bp. b, c Medulla oblongata sections from rats injected with hα-synuclein-AAVs and killed at 2 months (n = 4) were processed for fluorescent in situ hybridization coupled with immunofluorescence to detect WPRE mRNA (white), ChAT (green) and DAPI (blue). Representative images show ChAT-positive cells (b) in the absence (no white signal) of WPRE hybridization (c). Insets show higher magnification images of the right DMnX. Scale bars = 100 μm (large panels) and 50 μm (insets). d, e mRNA expression of WPRE (white) was evaluated in 4 rats killed 3 weeks after a single midbrain AAV injection. Midbrain sections were processed for fluorescent in situ hybridization and DAPI (blue). Representative images show right (AAV-injected) and left hemispheres (d, Bregma = −5.64 mm) or only the right hemisphere (e, Bregma = −4.92 mm). Scale bars = 1 mm. f Rats (n = 5) received midbrain injections of GFP-AAVs and were killed after 2 months. Non-injected (NI) animals provided control samples. GFP, WPRE, or hypoxantine phosphoribosyltransferase 1 (HPRT) mRNA was assayed by RT-PCR in samples of the right ventral mesencephalon (VM) or dorso-medial medulla oblongata (MO). Specific bands were at 94 (GFP), 204 (WPRE), and 61 (HPRT) bp.