Heterogeneous transgene expression in the retinas of the TH-RFP, TH-Cre, TH-BAC-Cre and DAT-Cre mouse lines.
Transgenic mouse lines are essential tools for understanding the connectivity, physiology and function of neuronal circuits, including those in the retina. This report compares transgene expression in the retina of a tyrosine hydroxylase (TH)-red fluorescent protein (RFP) mouse line with three catecholamine-related Cre recombinase mouse lines [TH-bacterial artificial chromosome (BAC)-, TH-, and dopamine transporter (DAT)-Cre] that were crossed with a ROSA26-tdTomato reporter line. Retinas were evaluated and immunostained with commonly used antibodies including those directed to TH, GABA and glycine to characterize the RFP or tdTomato fluorescent-labeled amacrine cells, and an antibody directed to RNA-binding protein with multiple splicing to identify ganglion cells. In TH-RFP retinas, types 1 and 2 dopamine (DA) amacrine cells were identified by their characteristic cellular morphology and type 1 DA cells by their expression of TH immunoreactivity. In the TH-BAC-, TH-, and DAT-tdTomato retinas, less than 1%, ∼ 6%, and 0%, respectively, of the fluorescent cells were the expected type 1 DA amacrine cells. Instead, in the TH-BAC-tdTomato retinas, fluorescently labeled AII amacrine cells were predominant, with some medium diameter ganglion cells. In TH-tdTomato retinas, fluorescence was in multiple neurochemical amacrine cell types, including four types of polyaxonal amacrine cells. In DAT-tdTomato retinas, fluorescence was in GABA immunoreactive amacrine cells, including two types of bistratified and two types of monostratified amacrine cells. Although each of the Cre lines was generated with the intent to specifically label DA cells, our findings show a cellular diversity in Cre expression in the adult retina and indicate the importance of careful characterization of transgene labeling patterns. These mouse lines with their distinctive cellular labeling patterns will be useful tools for future studies of retinal function and visual processing.
Molecular, Cellular, and Integrative Physiology Program, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States; Department of Neurobiology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States; Jules Stein Eye Institute, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States.
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Vuong
Department of Neurobiology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States.
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Pérez de Sevilla Müller
Department of Psychology, College of Letters and Science, UCLA, Los Angeles, CA 90095, United States.
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Hardi
Department of Biological Sciences, Vanderbilt University, Nashville, TN 37235, United States.
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McMahon
Molecular, Cellular, and Integrative Physiology Program, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States; Department of Neurobiology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States; Jules Stein Eye Institute, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States; CURE-Digestive Diseases Research Center, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, United States; Veterans Administration Greater Los Angeles Healthcare System, Los Angeles, CA 90095, United States. Electronic address: nbrecha@ucla.edu.
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Brecha
D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives / Q000378:metabolism; D064032:Calbindin 2 / Q000378:metabolism; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial; D050483:Dopamine Plasma Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics*; D005998:Glycine / Q000378:metabolism; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016601:RNA-Binding Proteins / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics / Q000378:metabolism*; D014795:Visual Pathways / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D064032:Calbindin 2; D050483:Dopamine Plasma Membrane Transport Proteins; D008164:Luminescent Proteins; D016601:RNA-Binding Proteins; C119265:Rbpms protein, mouse; C070614:neurobiotin; D005680:gamma-Aminobutyric Acid; D001710:Biotin; D014446:Tyrosine 3-Monooxygenase; D002795:Choline O-Acetyltransferase; C045073:Cre recombinase; D019426:Integrases; D005998:Glycine
Cre-lox-system; GABA; amacrine cells; dopamine; glycine; transgenic