PUBLICATION

Generation of Rab-based transgenic lines for in vivo studies of endosome biology in zebrafish

Authors
Clark, B.S., Winter, M., Cohen, A.R., and Link, B.A.
ID
ZDB-PUB-111024-9
Date
2011
Source
Developmental Dynamics : an official publication of the American Association of Anatomists   240(11): 2452-2465 (Journal)
Registered Authors
Clark, Brian, Link, Brian
Keywords
endosome dynamics, neuroepithelia, organelle polarization, zebrafish, transgenic lines
MeSH Terms
  • Models, Biological
  • Animals, Genetically Modified
  • Amino Acid Sequence
  • Animals
  • Sequence Homology, Amino Acid
  • Neuroepithelial Cells/metabolism
  • Endosomes/genetics
  • Endosomes/metabolism
  • Endosomes/physiology*
  • Biology/methods*
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zebrafish/physiology
  • rab5 GTP-Binding Proteins/genetics
  • rab5 GTP-Binding Proteins/metabolism
  • Membrane Proteins/metabolism
  • Gene Transfer Techniques*
  • Zebrafish Proteins/metabolism
  • rab GTP-Binding Proteins/genetics*
  • rab GTP-Binding Proteins/metabolism
(all 23)
PubMed
21976318 Full text @ Dev. Dyn.
Abstract
The Rab family of small GTPases function as molecular switches regulating membrane and protein trafficking. Individual Rab isoforms define and are required for specific endosomal compartments. To facilitate in vivo investigation of specific Rab proteins, and endosome biology in general, we have generated transgenic zebrafish lines to mark and manipulate Rab proteins. We also developed software to track and quantify endosome dynamics within time-lapse movies. The established transgenic lines ubiquitously express EGFP fusions of Rab5c (early endosomes), Rab11a (recycling endosomes), and Rab7 (late endosomes) to study localization and dynamics during development. Additionally, we generated UAS-based transgenic lines expressing constitutive active (CA) and dominant-negative (DN) versions for each of these Rab proteins. Predicted localization and functional consequences for each line were verified through a variety of assays, including lipophilic dye uptake and Crumbs2a localization. In summary, we have established a toolset for in vivo analyses of endosome dynamics and functions.
Genes / Markers
Figures
Figure Gallery (9 images)
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
No data available
Sequence Targeting Reagents
Target Reagent Reagent Type
rab5cMO3-rab5cMRPHLNO
rab11aMO2-rab11aMRPHLNO
1 - 2 of 2
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Fish
Antibodies
Orthology
Engineered Foreign Genes
Marker Marker Type Name
EGFPEFGEGFP
GAL4EFGGAL4
mCherryEFGmCherry
1 - 3 of 3
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Mapping
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Citation
Clark, B.S., Winter, M., Cohen, A.R., and Link, B.A. (2011) Generation of Rab-based transgenic lines for in vivo studies of endosome biology in zebrafish. Developmental Dynamics : an official publication of the American Association of Anatomists. 240(11):2452-2465.