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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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          Author and article information

          Journal
          101282001
          33781
          Brain Struct Funct
          Brain Struct Funct
          Brain structure & function
          1863-2653
          1863-2661
          11 October 2017
          12 November 2015
          September 2016
          26 October 2017
          : 221
          : 7
          : 3729-3741
          Affiliations
          [1 ]Department of Experimental Zoology and Neurobiology, University of Pécs, Pécs, Hungary
          [2 ]Laboratory of Molecular Biology and Genetics, Institute of Experimental Medicine, Hungarian Academy of Sciences, 1450 Budapest, Hungary
          [3 ]Center for Structural and Functional Neuroscience, University of Montana, Missoula, MT, USA
          [4 ]Department of Biomedical and Pharmaceutical Sciences, University of Montana, Missoula, MT, USA
          [5 ]Institute of Sport Sciences and Physical Education, University of Pécs, Ifjúság u. 6., 7624 Pécs, Hungary
          Author notes
          [6]

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

          Article
          PMC5657148 PMC5657148 5657148 nihpa910272
          10.1007/s00429-015-1128-2
          5657148
          26563404
          27ce5742-404b-4285-9a2c-c3a07f4dee4b
          History
          Categories
          Article

          GAD65-GFP,Expression mismatch,ChAT-tauGFP,ChAT-CRE/Rosa26YFP,PV-CRE/Rosa26YFP

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