In the SN, expression of hCDNF was delayed and clearly seen only after 12 weeks. The amount of expressed CDNF and GDNF proteins was similar to each other at 8–9 weeks after virus vector injection. Thus, it is obvious that the neuroprotection observed in this study is due to expression of the neurotrophic factors in the brain. The protection of TH-positive cells in the SN was rather modest. This may be related to the pattern

of protein expression and the titers used. Thus, Akt inhibitor analyses of the striata revealed a much more spatially restricted expression of hCDNF as compared with Inhibitors,research,lifescience,medical that of GDNF. This may explain why the protection of TH-reactive cells in the SNpc in AAV2-CDNF-treated rats was seen mainly in the central subdivision of the SN. In AAV2-GDNF-treated rats, protection of TH-reactive cells seemed to be Inhibitors,research,lifescience,medical more consistent across all the analyzed nigral sections, although the effect was statistically nonsignificant. TH-reactive cells from different anterior–posterior levels

of the rat SN have been reported to respond differently to both intrastriatal injections of 6-OHDA (Kirik et al. 1998) and to GDNF gene therapy (single intrastriatal injection of recombinant adenoviral vector) (Choi-Lundberg et al. 1998). Whether this is a consequence of the position of the striatal injection, spreading, Inhibitors,research,lifescience,medical and distribution of the neurotrophic factor or difference in the responsiveness of DAergic Inhibitors,research,lifescience,medical cells remains unclear. In a previous study, where approximately nine times bigger AAV2-GDNF titer dose was divided into three sites throughout the striatum intrastriatal AAV2-GDNF was shown to provide significant protection of nigral DAergic cells (Kirik et al. 2000). Therefore, it is likely that the titers used in this study were too low to provide maximal protection of the midbrain

DAergic neurons against 6-OHDA toxicity. Also, the transduction Inhibitors,research,lifescience,medical volume of AAV serotype 2 is known to be low, and other serotypes providing better spread of the viral transgene and increased expression (e.g., AAV serotype 5; Burger et al. 2004) would probably also result in better protection of the nigrostriatal pathway. On the other hand, AAV2-NRTN (CERE-120) provided significant protection of nigral TH-reactive cells even at viral vector doses as low as 1.6 × 108 vg (single-site secondly injection of 6-OHDA) (Gasmi et al. 2007a). Delivery of AAV2-GDNF prior to 6-OHDA administration provided an increase in TH-immunoreactive fiber density in the striatum, and sprouting of TH-immunoreactive fibers in the lateral GP and SNpr, as reported also by others (Kirik et al. 2000; Kordower et al. 2000; Georgievska et al. 2002a,b). We did not observe any sprouting of TH-positive fibers in the striatum of AAV2-CDNF-treated animals, even though the fiber density was partly preserved.