A versatile system for the neuronal subtype specific expression of lentiviral vectors

Abstract

Lentiviral expression vectors are powerful tools for gene therapy and long-term gene expression/repression in the mammalian brain. However, no specificity of transduction has been reported so far in the central nervous system. Here we have developed a novel system to achieve a neuronal subtype specific expression in either dopaminergic (DA) or GABAergic neurons. We employed a delivery strategy by which the transgene is not expressed until its activation by Cre recombinase. We successfully tested the system in vitro and then used this novel lentivector, containing loxP sites, in 2 different transgenic mouse lines expressing Cre either in DA or in GABAergic neurons. In both lines the reporter gene was detected exclusively in Cre-positive cells, demonstrating that with this experimental approach we were able to achieve completely specific expression of transgenes delivered by lentiviral vectors. This universal system can be applied to all neural subtypes making use of the growing number of specific Cre driver lines.- Tolu, S., Avale, M. E., Nakatani, H., Pons, S., Parnaudeau, S., Tronche, F., Vogt, A., Monyer, H., Vogel, R., de Chaumont, F., Olivo-Marin, J.-C., Changeux, J.-P., Maskos, U. A versatile system for the neuronal subtype specific expression of lentiviral vectors.

Nicotine consumption is regulated by a human polymorphism in dopamine neurons

“…Thereafter, to address and ascertain the specific role of α5*-nAChRs in VTA DA cells, a DA cell-specific expression system was generated similar to Tolu et al 14,22 α5 − / − -DAT Cre mice were generated by crossing α5 − / − with DAT-Cre expressing transgenic mice. These mice were injected with a Cre recombinasedependent conditional lentiviral expression vector to drive α5WT or α5SNP exclusively in VTA DA neurons (Figure 2a, bottom).…”

“…The lentiviral expression vectors are derived from the pHR's expression vectors first described by Naldini et al, 20 with several subsequent modifications. 19,21,22 To create the conditional lentivectors, a previously described sub-cloning strategy was used. 14,19,21,22 Data analysis Behavioral data.…”

“…19,21,22 To create the conditional lentivectors, a previously described sub-cloning strategy was used. 14,19,21,22 Data analysis Behavioral data. The number of nose pokes (NPs) for both A and P mice in each treatment group was analyzed, first with a Shapiro test, then with two-way analysis of variance to evaluate effects of the drug delivery mode, unit dose and interactions between group and drug dose.…”

GFRα1 Regulates Purkinje Cell Migration by Counteracting NCAM Function

“…The following transgenic mouse lines were used for experiments: Gfra1 KO (Enomoto et al., 1998), Gdnf KO (Pichel et al., 1996), Gdnf bgal (Moore et al., 1996), Ret EGFP (Jain et al., 2006), Gfra1 fx (kindly provided by M. Saarma and J.-O. Andressoo, University of Helsinki), Ptf1a Cre (Kawaguchi et al., 2002), Gad67 Cre (Tolu et al., 2010), Nestin Cre (Tronche et al., 1999), Rosa26 dTom (Madisen et al., 2010), Gfra1 EGFP (Uesaka et al., 2007), and Gfra1 CreERT2 (this study). The latter were generated at TaconicArtemis by inserting a CreERT2 cassette in frame with the second exon of the Gfra1 gene (Figure S2).…”

Protein O-Mannosylation in the Murine Brain: Occurrence of Mono-O-Mannosyl Glycans and Identification of New Substrates

“…As shown in Fig 3A, the staining overlapped to a high extent in the hippocampus (upper panels) and cerebellum (lower panels), although additional α-O-Man epitopes were detected for example in the granular cell layer (see also inlay in lower panel 3). Moreover, in brains from transgenic mice expressing the cre protein in GABAergic neurons [44] perfect co-localization of α-O-Man immunoreactivity with cre-positve neurons was observed. (Fig 3B), further corroborating the pronounced occurrence of mono-O-mannosyl glycans in GABAergic inhibitory neurons.…”