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Ectopic transgene expression in the retina of four transgenic mouse lines

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Abstract

Retinal expression of transgenes was examined in four mouse lines. Two constructs were driven by the choline acetyltransferase (ChAT) promoter: green fluorescent protein conjugated to tau protein (tau-GFP) or cytosolic yellow fluorescent protein (YFP) generated through CRE recombinase-induced expression of Rosa26 (ChAT-CRE/Rosa26YFP). Two other constructs targeted inhibitory interneurons: GABAergic horizontal and amacrine cells identified by glutamic acid decarboxylase (GAD65-GFP) or parvalbumin (PV) cells (PV-CRE/Rosa26YFP). Animals were transcardially perfused and retinal sections prepared. Antibodies against PV, calretinin (CALR), calbindin (CALB), and tyrosine hydroxylase (TH) were used to counterstain transgene-expressing cells. In PVxRosa and ChAT-tauGFP constructs, staining appeared in vertically oriented row of processes resembling Müller cells. In the ChATxRosa construct, populations of amacrine cells and neurons in the ganglion cell layer were labeled. Some cones also exhibited GFP fluorescence. CALR, PV and TH were found in none of these cells. Occasionally, we found GFP/CALR and GFP/PV double-stained cells in the ganglion cell layer (GCL). In the GAD65-GFP construct, all layers of the neuroretina were labeled, except photoreceptors. Not all horizontal cells expressed GFP. We did not find GFP/TH double-labeled cells and GFP was rarely present in CALR- and CALB-containing cells. Many PV-positive neurons were also labeled for GFP, including small diameter amacrines. In the GCL, single labeling for GFP and PV was ascertained, as well as several CALR/PV double-stained neurons. In the GCL, cells triple labeled with GFP/CALR/CALB were sparse. In conclusion, only one of the four transgenic constructs exhibited an expression pattern consistent with endogenous retinal protein expression, while the others strongly suggested ectopic gene expression.

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Acknowledgments

This study was supported by NAP KTIA_13_NAP-A-I/12 (R.G.) and NIH R01 NS069689 (J.J.L.) Grants. M.W. was in receipt of a short-term fellowship from the College of Health Professions and Biomedical Sciences, University of Montana. R.G. was a Fulbright Fellow in this institution. This project was also supported through Grants from the Center for Environmental Health Sciences COBRE P20RR017670, Center for Biomolecular Structure and Dynamics P20GM103546, COBRE Center for Structural and Functional Neuroscience P20RR015583, and an internal Grant from the University of Montana, Department of Biomedical Sciences. We thank Feng Yi and Elizabeth Catudio-Garrett for help with the transgenic animals, Sukumar Vijayaraghavan (UC-Denver) for ChAT-tauGFP mice, and Edit Kiss with the preparation of the figures.

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  1. J. Josh Lawrence

    Present address: Department of Pharmacology and Neuroscience, Texas Tech University Health Sciences Center, School of Medicine, Lubbock, TX, 79430, USA

Authors and Affiliations

  1. Department of Experimental Zoology and Neurobiology, University of Pécs, Pécs, Hungary

    Robert Gábriel

  2. Laboratory of Molecular Biology and Genetics, Institute of Experimental Medicine, Hungarian Academy of Sciences, 1450, Budapest, Hungary

    Ferenc Erdélyi & Gábor Szabó

  3. Center for Structural and Functional Neuroscience, University of Montana, Missoula, MT, USA

    J. Josh Lawrence

  4. Department of Biomedical and Pharmaceutical Sciences, University of Montana, Missoula, MT, USA

    J. Josh Lawrence

  5. Institute of Sport Sciences and Physical Education, University of Pécs, Ifjúság u. 6., 7624, Pécs, Hungary

    Márta Wilhelm

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  1. Robert Gábriel
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Correspondence to Márta Wilhelm.

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Gábriel, R., Erdélyi, F., Szabó, G. et al. Ectopic transgene expression in the retina of four transgenic mouse lines. Brain Struct Funct 221, 3729–3741 (2016). https://doi.org/10.1007/s00429-015-1128-2

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  • DOI: https://doi.org/10.1007/s00429-015-1128-2

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