Zebrafish Tg(7.2mab21l2:EGFP)ucd2 Transgenics Reveal a Unique Population of Retinal Amacrine Cells

Cederlund, M.L., Morrissey, M.E., Baden, T., Scholz, D., Vendrell, V., Lagnado, L., Connaughton, V.P., and Kennedy, B.N.
Investigative ophthalmology & visual science   52(3): 1613-21 (Journal)
Registered Authors
Cederlund, Maria, Connaughton, Victoria P., Kennedy, Breandan N., Lagnado, Leon
amacrine cell, transgenic animals, electrophysiology, confocal microscopy, development, ocular
MeSH Terms
  • Amacrine Cells/cytology*
  • Amacrine Cells/metabolism
  • Animals
  • Animals, Genetically Modified
  • Calbindin 2
  • Electrophysiology
  • Embryo, Nonmammalian/cytology*
  • 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*
  • gamma-Aminobutyric Acid/metabolism
21051702 Full text @ Invest. Ophthalmol. Vis. Sci.
Purpose: Amacrine cells comprise a diverse, yet poorly characterised, cell population in the inner retina. Here, we seek to characterise the morphology, molecular physiology and electrophysiology of a subpopulation of EGFP expressing retinal amacrine cells identified in a novel zebrafish transgenic line. Methods: 7.2 kb of the zebrafish mab21l2 promoter was cloned upstream of EGFP and used to create the Tg(7.2mab21l2:EGFP)ucd2 transgenic line. Transgenic EGFP expression was analysed by fluorescent microscopy in wholemount embryos followed by detailed analysis of EGFP-expressing amacrine cells using fluorescent microscopy, immunohistochemistry and electrophysiology. Results: 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-labelled amacrine cells in Tg(7.2mab21l2:EGFP)ucd2 constitute a novel GABA- and glycine-negative amacrine subpopulation. Morphologically, EGFP-expressing cells stratify in sublamina 1-2 (Type 1 OFF), or sublamina 3-4 (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.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes