Abstract
BackgroundTransgenic 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 FindingsHere 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.SignificanceThis triple transgenic approach will aid the morphological and functional characterization of neurons in various Cre-dependent transgenic mice.
Highlights
The development and functionality of the central nervous system involves complex interactions between various neuronal cell types
The advent of green fluorescent protein (GFP) as a tool to label cells [2] together with live cell imaging by confocal- [3,4,5,6] or two photon microscopy [7,8,9,10] have permitted the study of dynamic structural changes occurring with development and plasticity of the central nervous system in vivo [11,12,13]
In vivo two–photon imaging of transgenic mice with mosaic, Golgi-staining like GFP expression has made the analysis of the dynamics of structural changes in neurons possible
Summary
The development and functionality of the central nervous system involves complex interactions between various neuronal cell types. For more than a century, the majority of morphological studies in neuroscience at the cellular and subcellular level have been carried out using the Golgi approach to stain individual neurons [1] and visualize their dendrites and dendritic protrusions. This approach has been vital for the elucidation of the structural details of the central nervous system, it is not suitable for examining the dynamic functions of neurons. It has been difficult to obtain reproducible expression in the same subset of neurons for direct comparison of neurons expressing different functional proteins
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