Establishment of CNS-related genetic tools
1. Genetic approach for intracerebroventricular delivery
Administration of synthetic or purified peptides directly into the brain ventricles is a method commonly used by neuroscientists for exploring physiological and behavioral functions of gene products. Intravenous administration is controlled by the blood-brain barrier, which limits its effectiveness, and current approaches for acute or chronic intra-cerebro-ventricular delivery have significant technical drawbacks resulting from both the chemical properties of the delivered substance, and the experimental procedures. Here we describe a genetic approach for the delivery of secreted peptides or proteins into the cerebrospinal fluid (CSF). Using a choroid plexus-specific promoter, we established a lentiviral-based system, which offers inducible and reversible delivery of a gene product into the CSF. The functionality of this system was demonstrated using the over-expression of the two established neuropeptides, corticotropin-releasing factor and gonadotropin-releasing hormone, modulating anxiety-like behavior and estrus cycle, respectively. We show that this choroid plexus-specific lentiviral-based system is a reliable, effective and adaptable research tool for intra-cerebro-ventricular delivery. (Regev et al., Genetic approach for intracerebroventricular delivery. Proc Natl Acad Sci U S A 107:4424-4429, 2010)
Schematic representation of a lentiviral-based system designed for inducible over-expression of peptides or proteins of interest by the choroid plexus cells. rtTA, reverse tetracycline trans activator; TRE, tetracycline-responsive element; ISF, interstitial fluid. (For further information on this figure, please refer to the article by Regev et al., Proc Natl Acad Sci U S A 107:4424-4429, 2010).
