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In vivo gene silencing of CD81 by lentiviral expression of small interference RNAs suppresses cocaine‐induced behaviour
Author(s) -
Bahi Amine,
Boyer Frederic,
Kolira Manoj,
Dreyer JeanLuc
Publication year - 2005
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2004.02961.x
Subject(s) - cd81 , gene silencing , small hairpin rna , gene knockdown , rna interference , biology , nucleus accumbens , microbiology and biotechnology , ventral tegmental area , dopaminergic , rna , gene , receptor , dopamine , immunology , biochemistry , neuroscience , virus , hepatitis c virus
The tetraspanin CD81 is induced in the mesolimbic dopaminergic pathway after cocaine administration. To further investigate its role, a regulatable lentivirus (Lenti‐CD81) bearing the CD81 gene under the control of a tetracycline‐inducible promoter and lentiviruses expressing short hairpin RNA (shRNA) targeted against CD81 (Lenti‐CD81‐shRNAs) have been prepared. Infection of HEK293T cells in vitro with Lenti‐CD81‐shRNAs resulted in 96.5% gene silencing (from quantitative real‐time PCR(qRT‐PCR) and immunocytochemistry). In vivo delivery of Lenti‐CD81‐shRNA into the nucleus accumbens or ventral tegmental area resulted in 91.3 and 94% silencing of endogenous CD81, respectively. Stereotaxic injection of Lenti‐CD81 into these regions, resulting in CD81 overexpression, induced a four‐ to fivefold increase in locomotor activity after chronic cocaine administration, which returned to basal levels when Lenti‐CD81‐shRNA had been coinjected or when CD81 expression was blocked by doxycycline. Furthermore, silencing endogenous CD81 in vivo resulted in a significant decrease in locomotor activity over controls, again suppressing cocaine‐induced behaviour.