Zebrafish Tg(7.2mab21l2:EGFP)ucd2 transgenics reveal a unique population of retinal amacrine cells

Amacrine cells constitute a diverse, yet poorly characterized, cell population in the inner retina. Here, the authors sought to characterize the morphology, molecular physiology, and electrophysiology of a subpopulation of EGFP-expressing retinal amacrine cells identified in a novel zebrafish transg...

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Bibliographic Details
Published in:Investigative ophthalmology & visual science 2011-03, Vol.52 (3), p.1613-1621
Main Authors: Cederlund, Maria L, Morrissey, Maria E, Baden, Tom, Scholz, Dimitri, Vendrell, Victor, Lagnado, Leon, Connaughton, Victoria P, Kennedy, Breandán N
Format: Article
Language:English
Subjects:
Amacrine Cells - cytology
Amacrine Cells - metabolism
Animals
Animals, Genetically Modified
Calbindin 2
Electrophysiology
Embryo, Nonmammalian - cytology
gamma-Aminobutyric Acid - metabolism
Gene Expression Regulation, Developmental - physiology
Glycine - metabolism
Green Fluorescent Proteins - genetics
Homeodomain Proteins - genetics
Immunohistochemistry
Microscopy, Fluorescence
Parvalbumins - metabolism
Promoter Regions, Genetic
Retina - embryology
Retina - metabolism
S100 Calcium Binding Protein G - metabolism
Zebrafish
Zebrafish Proteins - genetics
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Summary:Amacrine cells constitute a diverse, yet poorly characterized, cell population in the inner retina. Here, the authors sought to characterize the morphology, molecular physiology, and electrophysiology of a subpopulation of EGFP-expressing retinal amacrine cells identified in a novel zebrafish transgenic line. After 7.2 kb of the zebrafish mab21l2 promoter was cloned upstream of EGFP, it was used to create the Tg(7.2mab21l2:EGFP)ucd2 transgenic line. Transgenic EGFP expression was analyzed by fluorescence microscopy in whole mount embryos, followed by detailed analysis of EGFP-expressing amacrine cells using fluorescence microscopy, immunohistochemistry, and electrophysiology. A 7.2-kb fragment of the mab21l2 promoter region is sufficient to drive transgene expression in the developing lens and tectum. Intriguingly, EGFP was also observed in differentiated amacrine cells. EGFP-labeled amacrine cells in Tg(7.2mab21l2:EGFP)ucd2 constitute a novel GABA- and glycine-negative amacrine subpopulation. Morphologically, EGFP-expressing cells stratify in sublamina 1 to 2 (type 1 OFF) or sublamina 3 to 4 (type 1 ON) or branch diffusely (type 2). Electrophysiologically, these cells segregate into amacrine cells with somas in the vitreal part of the INL and linear responses to current injection or, alternatively, amacrine cells with somas proximal to the IPL and active oscillatory voltage signals. CONCLUSIONS; The novel transgenic line Tg(7.2mab21l2:EGFP)ucd2 uncovers a unique subpopulation of retinal amacrine cells.
ISSN:1552-5783
0146-0404
1552-5783
DOI:10.1167/iovs.10-5376