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


RESULTS
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.


CONCLUSIONS
This triple transgenic approach will aid the morphological and functional characterization of neurons in various Cre-dependent transgenic mice.
Chakravarthy
Sridhara
S
Molecular Visual Plasticity Group, Netherlands Institute for Neuroscience, Amsterdam, The Netherlands.
Keck
Tara
T
Roelandse
Martijn
M
Hartman
Robin
R
Jeromin
Andreas
A
Perry
Sean
S
Hofer
Sonja B
SB
Mrsic-Flogel
Thomas
T
Levelt
Christiaan N
CN
D000328:Adult; D000818:Animals; D001921:Brain / Q000502:physiology; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009030:Mosaicism; D009474:Neurons / Q000502:physiology*; D016548:Prosencephalon / Q000502:physiology*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases