In vivo gene silencing of CD81 by lentiviral expression of small interference RNAs suppresses cocaine-induced behaviour
- Bahi, Amine Institute of Biochemistry, University of Fribourg, Fribourg, Switzerland
- Boyer, Frederic Institute of Biochemistry, University of Fribourg, Fribourg, Switzerland
- Kolira, Manoj Institute of Biochemistry, University of Fribourg, Fribourg, Switzerland
- Dreyer, Jean-Luc Institute of Biochemistry, University of Fribourg, Fribourg, Switzerland
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04.02.2005
Published in:
- Journal of Neurochemistry. - 2005, vol. 92, no. 5, p. 1243-1255
English
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.
- Faculty
- Faculté des sciences et de médecine
- Department
- Département de Médecine
- Language
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- English
- Classification
- Biological sciences
- License
- License undefined
- Identifiers
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- RERO DOC 5127
- DOI 10.1111/j.1471-4159.2004.02961.x
- Persistent URL
- https://folia.unifr.ch/unifr/documents/299853
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