Adeno-associated virus-mediated ILK gene silencing in the rat NAc core

In this study we established conditional silencing of integrin-linked kinase (ILK) expression in Sprague-Dawley rat brain by microinjection of rAAV-2-carrying, Tet-On-regulated siRNA expression cassette into nucleus accumbens (NAc) core and induction with doxycycline. We demonstrated that inhibition of ILK expression was effectively induced by administration of doxycycline for 2 weeks while ILK expression was restored after withdrawing doxycycline for 8 days. Increases in GFAP and OX42 expression were observed 5 weeks post virus injection. Importantly, inhibition of ILK expression in the NAc core had no significant effect on cell apoptosis and animal basal locomotion and stereotypical behaviors, but decreased dendritic density of medium spiny neurons. Our studies suggest that: (1) rAAV-delivered Tet-On-regulated siRNA expression can conditionally regulate gene expression in rat brain; (2) inhibition of ILK expression has no significant effect on cell apoptosis and basal locomotor and stereotypical behaviors, but decreases dendritic density; and (3) microinjection of rAAV-2 causes inflammatory response around the injection track. (c) 2008 Elsevier B.V. All rights reserved.