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Accepted (AM)Open Access (License Unspecified), Open
Journal article
Generation of Rab-Based Transgenic Lines for In Vivo Studies of Endosome Biology in Zebrafish
Developmental dynamics, v 240(11), pp 2452-2465
01 Nov 2011
PMID: 21976318
Featured in Collection : UN Sustainable Development Goals @ Drexel
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. Developmental Dynamics 240: 2452-2465, 2011. (C) 2011 Wiley Periodicals, Inc.
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Details
- Title
- Generation of Rab-Based Transgenic Lines for In Vivo Studies of Endosome Biology in Zebrafish
- Creators
- Brian S. Clark - Medical College of WisconsinMark Winter - University of Wisconsin–MilwaukeeAndrew R. Cohen - University of Wisconsin–MilwaukeeBrian A. Link - Medical College of Wisconsin
- Publication Details
- Developmental dynamics, v 240(11), pp 2452-2465
- Publisher
- Wiley
- Number of pages
- 14
- Grant note
- E. Matilda Ziegler Foundation R01NS076709 / NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Neurological Disorders & Stroke (NINDS) P30EY001931 / National Eye Institute; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Eye Institute (NEI) R01EY014167 / NATIONAL EYE INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Eye Institute (NEI) T32EY014536; R01NS076709; R01EY014167 / National Institutes of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Electrical and Computer Engineering
- Web of Science ID
- WOS:000296971700005
- Scopus ID
- 2-s2.0-80054832731
- Other Identifier
- 991019297215504721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Anatomy & Morphology
- Developmental Biology