title: Adeno-associated virus-mediated ILK gene silencing in the rat NAc core.
abstract: 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.
authors.affiliation: Department of Psychiatry and Behavioral Sciences, Duke University Medical Center, PO Box 3870, Research Drive, Durham, NC 27710, USA.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
authors.forename: Xueying
authors.initials: X
authors.lastname: Xiong
authors.forename: Tong H
authors.initials: TH
authors.lastname: Lee
authors.forename: Yingmiao
authors.initials: Y
authors.lastname: Liu
authors.forename: Qi-An
authors.initials: QA
authors.lastname: Sun
authors.forename: William
authors.initials: W
authors.lastname: Wetsel
authors.forename: Xiuwu
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D049229:Dendritic Spines / Q000502:physiology / Q000648:ultrastructure; D000229:Dependovirus / Q000235:genetics; D004318:Doxycycline / Q000494:pharmacology; D004660:Encephalitis / Q000139:chemically induced / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing / Q000502:physiology*; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D008297:Male; D008845:Microinjections / Q000009:adverse effects; D008967:Molecular Biology / Q000379:methods*; D009043:Motor Activity / Q000187:drug effects / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000502:physiology; D009714:Nucleus Accumbens / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D034622:RNA Interference / Q000502:physiology; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013239:Stereotyped Behavior / Q000187:drug effects / Q000502:physiology; D000099200:Scaffold Protein ILK
chemical_list: D000900:Anti-Bacterial Agents; D004318:Doxycycline; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D000099200:Scaffold Protein ILK