Expression analysis of green fluorescent protein in retinal neurons of four transgenic mouse lines

doi:10.1016/j.neuroscience.2009.01.081

Neuroscience

Volume 160, Issue 1, 21 April 2009, Pages 126-139

https://doi.org/10.1016/j.neuroscience.2009.01.081 Get rights and content

Abstract

Transgenic mice that express enhanced green fluorescent protein (EGFP) under the control of a cell-specific promoter have been used with great success to identify and label specific cell types of the retina. We studied the expression of EGFP in the retina of mice making use of four transgenic mouse lines. Expression of EGFP driven by the calretinin promoter was found in amacrine, displaced amacrine and ganglion cells. Comparison of the EGFP expression and calretinin immunolabeling showed that many but not all cells appear to be double labeled. Expression of EGFP under the control of the choline acetyltransferase promoter was found in amacrine cells; however, the cells did not correspond to the well known cholinergic (starburst) cells of the mouse retina. The expression of EGFP under the control of the parvalbumin promoter was restricted to amacrine cells of the inner nuclear layer and to cells of the ganglion cell layer (displaced amacrine cells and ganglion cells). Most of the cells were also immunoreactive for parvalbumin, however, differences in labeling intensity were observed. The expression of EGFP driven by the promoter for the 5-HT3 A receptor (5-HTR3A) was restricted to type 5 bipolar cells. In contrast, immunostaining for 5-HTR3A was found in synaptic hot spots in sublamina 1 of the inner plexiform layer and was not related to type 5 bipolar cells. The results show that these transgenic mice are very useful for future electrophysiological studies of specific types of amacrine and bipolar cells that express EGFP and thus permit directed microelectrode targeting under microscopic control.

Section snippets

Generation of transgenic mice

The generation of the PV-EGFP transgenic mice was described in detail by Meyer et al. (2002), the generation of the ChAT-EGFP transgenic mice was described by von Engelhardt et al. (2007), and CR-EGFP transgenic mice were described by Caputi et al. (2008).

5-HTR3A-EGFP transgenic mice were generated using BACs by homologous recombination (Inta et al., 2008). BAC clones spanning the mouse 5-HTR3A gene were identified by polymerase chain reaction screening of libraries of mouse genomic DNA.

Results

In all four transgenic mouse lines a specific expression pattern of EGFP could be observed (Fig. 1). In the mouse line (Fig. 1A), where EGFP expression was under the control of the CR promoter (CR-EGFP mouse) label was confined to amacrine cells of the inner nuclear layer (INL) and to displaced amacrine cells and possibly ganglion cells of the GCL. Two narrow dendritic strata were labeled in the IPL.

The labeling pattern of the ChAT-EGFP mouse (Fig. 1B) was different. Many amacrine cells of the

The BAC technology

Transgenic mice that express reliably and at high level detectable markers such as EGFP in distinct neuronal populations represent an important tool for anatomical and functional studies. The generation of transgenic mice using BACs has become a very useful approach for studying gene expression in vivo in the nervous system (Gong et al., 2003). Generally, BAC transgenes allow the labeling of entire neuronal populations. Nevertheless, they do not guarantee a perfect recapitulation of the

Acknowledgments

We thank B. Marshallsay for excellent technical assistance and I. Odenthal for typing the manuscript.

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NeuroanatomyResearch PaperExpression analysis of green fluorescent protein in retinal neurons of four transgenic mouse lines

Abstract

Section snippets

Generation of transgenic mice

Results

The BAC technology

Acknowledgments

Brain Res

Vis Res

Neuron

Neuron

Neuron

Brain Res

Neuron

Neuron

Mol Brain Res

Exp Eye Res

Neuroscience

Neuroscience

J Neurosci Methods

Mol Brain Res

Vis Res

Cell

Neuron

Cell Calcium

Quantitative analysis of neuronal morphologies in the mouse retina visualized by using a genetically directed reporter

J Comp Neurol

Imaging synaptic inhibition in transgenic mice expressing the chloride indicator, clomeleon

Brain Cell Biol

Localization of biologically active peptides in the retina

Neuropharmacological analysis of the role of indoleamine-accumulating amacrine cells in the rabbit retina

Vis Neurosci

A role for 5HT3 receptors in visual processing in the mammalian retina

Vis Neurosci

Two calretinin-positive GABAergic cell types in layer 2/3 of the mouse neocortex provide different forms of inhibition

Cereb Cortex

The 5-HT3B subunit is a major determinant of serotonin-receptor function

Nature

5-HT3 receptors are membrane ion channels

Nature

Probing neurochemical structure and function of retinal ON bipolar cells with a transgenic mouse

J Comp Neurol

Ganglion cell distribution in the retina of the mouse

Invest Ophthalmol Vis Sci

Ionotropic glutmate receptors of amacrine cells of the mouse retina

Vis Neurosci

Indoleamine accumulating neurons in the retina of rabbit, cat, and goldfish

Cell Tissue Res

Transgenic expression of the jellyfish green fluorescent protein in the cone photoreceptors of the mouse

Vis Neurosci

Types of bipolar cells in the mouse retina

J Comp Neurol

A gene expression atlas of the central nervous system based on bacterial artificial chromosomes

Nature

Immunocytochemical analysis of the mouse retina

J Comp Neurol

Neuroanatomy
Research Paper
Expression analysis of green fluorescent protein in retinal neurons of four transgenic mouse lines