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Cationic lipid-mediated delivery and expression of prepro-neuropeptide Y cDNA after intraventricular administration in rat: feasibility and limitations.
Magnus Huss Clinic, Karolinska Hospital, S-17176 Stockholm, Sweden.
Department of Medical Genetics, Uppsala University, Uppsala, Sweden.
Magnus Huss Clinic, Karolinska Hospital, S-17176 Stockholm, Sweden.
1996 (English)In: Regulatory Peptides, ISSN 0167-0115, E-ISSN 1873-1686, Vol. 61, no 3, 205-211 p.Article in journal (Refereed) Published
Abstract [en]

The utility of in vivo lipofection for delivery and expression of a neuropeptide gene in the adult rat brain was explored. Prepro-neuropeptide Y (NPY) cDNA was cloned into the episomal eucaryotic expression vector pCEP4. This construct was complexed to lipofectamine or lipofectin. Complexed DNA was injected into the lateral ventricles of adult rats. Brains were removed for analysis following various time intervals. Polymerase chain reaction (PCR) reactions were designed for specific detection of endogenous and vector derived NPY sequence, respectively. PCR of DNA preparations from 5 major brain regions (frontal and parietal cortex, striatum, hypothalamus, brain stem) demonstrated presence of vector DNA up to 1 month (longest interval studied) in all brain regions. Reverse-transcription (RT-) PCR of DNase treated RNA-preparations from brain tissue demonstrated presence of both vector-derived and endogenous NPY mRNA in treated animals, while only endogenous mRNA was detected in controls. In situ hybridization histochemistry indicated scattered patches of vector uptake into tissue in the vicinity of the CSF compartment, but not into deeper located structures. Weight gain was not affected, indicating that the expression levels achieved may not be sufficient to play a functional role, and/or may need to be targeted to specific brain areas. These findings suggest a potential for cationic lipid mediated gene transfer in the brain as an experimental tool and as a possible future therapeutic principle, but also indicate the need for optimization of delivery strategies in order to achieve functionally relevant expression levels.

Place, publisher, year, edition, pages
1996. Vol. 61, no 3, 205-211 p.
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Neurosciences
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URN: urn:nbn:se:liu:diva-112519PubMedID: 8701037OAI: oai:DiVA.org:liu-112519DiVA: diva2:799377
Available from: 2015-03-30 Created: 2014-12-01 Last updated: 2017-12-04

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