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Cre-dependent expression of multiple transgenes in isolated neurons of the adult forebrain

Chakravarthy, S; Keck, T; Roelandse, M; Hartman, R; Jeromin, A; Perry, S; Hofer, SB; ... Levelt, CN; + view all (2008) Cre-dependent expression of multiple transgenes in isolated neurons of the adult forebrain. PLOS ONE , 3 (8) , Article e3059. 10.1371/journal.pone.0003059. Green open access

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Abstract

Background: Transgenic mice with mosaic, Golgi-staining-like expression of enhanced green fluorescent protein (EGFP) have been very useful in studying the dynamics of neuronal structure and function. In order to further investigate the molecular events regulating structural plasticity, it would be useful to express multiple proteins in the same sparse neurons, allowing co-expression of functional proteins or co-labeling of subcellular compartments with other fluorescent proteins. However, it has been difficult to obtain reproducible expression in the same subset of neurons for direct comparison of neurons expressing different functional proteins. Principal Findings: Here we describe a Cre-transgenic line that allows reproducible expression of transgenic proteins of choice in a small number of neurons of the adult cortex, hippocampus, striatum, olfactory bulb, subiculum, hypothalamus, superior colliculus and amygdala. We show that using these Cre-transgenic mice, multiple Cre-dependent transgenes can be expressed together in the same isolated neurons. We also describe a Cre-dependent transgenic line expressing a membrane associated EGFP (EGFP-F). Crossed with the Cre-transgenic line, EGFP-F expression starts in the adolescent forebrain, is present in dendrites, dendritic protrusions, axons and boutons and is strong enough for acute or chronic in vivo imaging. Significance: This triple transgenic approach will aid the morphological and functional characterization of neurons in various Cre-dependent transgenic mice.

Type: Article
Title: Cre-dependent expression of multiple transgenes in isolated neurons of the adult forebrain
Open access status: An open access version is available from UCL Discovery
DOI: 10.1371/journal.pone.0003059
Publisher version: http://dx.doi.org/10.1371/journal.pone.0003059
Language: English
Additional information: © 2008 Chakravarthy et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. S.C. was supported by Rotterdamse Vereniging Blindenbelangen, Algemeen Nederlandse Vereeniging ter Voorkoming van Blindheid and Stichting Blindenhulp. M.R. is supported by KWF Kanker Bestijding (NORI 2003-2789). C.N.L. is supported by the Royal Netherlands Academy of Arts and Sciences (K.N.A.W.), A VIDI grant from the Netherlands Organization for Scientific Research (N.W.O.), and a “Bsik” grant from SenterNovem. None of the funders were involved either in the design and implementation of the experiments or in the preparation of the manuscript.
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > The Sainsbury Wellcome Centre
URI: https://discovery.ucl.ac.uk/id/eprint/1366110
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