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.
Chen
Qiang
Q
Department of Psychiatry and Behavioral Sciences, Duke University Medical Center, PO Box 3870, Research Drive, Durham, NC 27710, USA.
Xiong
Xueying
X
Lee
Tong H
TH
Liu
Yingmiao
Y
Sun
Qi-An
QA
Wetsel
William
W
Zhang
Xiuwu
X
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
D000900:Anti-Bacterial Agents; D004318:Doxycycline; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D000099200:Scaffold Protein ILK