Live imaging of collagen deposition during skin development and repair in a collagen I – GFP fusion transgenic zebrafish line

Developmental biology

Yongbo Lu^4,5¹  Karl E. Kadler^3²  Josephine L. Morris^1³  Paul Martin^1,6,7⁴  Sarah L. Dallas^4⁵  Yinhui Lu^3⁶  Stephen J. Cross^2⁷ 

[1] Department of Biomedical Sciences, Texas A&M University College of Dentistry, 3302 Gaston Ave., Dallas, TX 75246, United States^5;Department of Oral and Craniofacial Sciences, School of Dentistry, University of Missouri-Kansas City, Kansas City, Missouri, United States^4;School of Biochemistry, Faculty of Biomedical Sciences, University Walk, University of Bristol, Bristol, BS8 1TD, UK^1;School of Medicine, Cardiff University, Cardiff, CF14 4XN, UK^7;School of Physiology, Pharmacology and Neuroscience, Faculty of Biomedical Sciences, University Walk, University of Bristol, Bristol, BS8 1TD, UK^6;Wellcome Trust Centre for Cell-Matrix Research, Faculty of Biology, Medicine and Health, The University of Manchester, Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK^3;Wolfson Bioimaging Facility, Faculty of Biomedical Sciences, University Walk, University of Bristol, Bristol, BS8 1TD, UK^2

关键词: Collagen-I;    Zebrafish larvae;    Live imaging;    Skin;    Wound healing;   

DOI  :  10.1016/j.ydbio.2018.06.001

学科分类：生物科学（综合）

来源: Academic Press

【 摘 要 】

Fibrillar collagen is a major component of many tissues but has been difficult to image in vivo using transgenic approaches because of problems associated with establishing cells and organisms that generate GFP-fusion collagens that can polymerise into functional fibrils. Here we have developed and characterised GFP and mCherry collagen-I fusion zebrafish lines with basal epidermal-specific expression. We use these lines to reveal the dynamic nature of collagen-I fibril deposition beneath the developing embryonic epidermis, as well as the repair of this collagen meshwork following wounding. Transmission electron microscope studies show that these transgenic lines faithfully reproduce the collagen ultrastructure present in wild type larval skin. During skin development we show that collagen I is deposited by basal epidermal cells initially in fine filaments that are largely randomly orientated but are subsequently aligned into a cross-hatch, orthogonal sub-epithelial network by embryonic day 4. Following skin wounding, we see that sub-epidermal collagen is re-established in the denuded domain, initially as randomly orientated wisps that subsequently become bonded to the undamaged collagen and aligned in a way that recapitulates developmental deposition of sub-epidermal collagen. Crossing our GFP-collagen line against one with tdTomato marking basal epidermal cell membranes reveals how much more rapidly wound re-epithelialisation occurs compared to the re-deposition of collagen beneath the healed epidermis. By use of other tissue specific drivers it will be possible to establish zebrafish lines to enable live imaging of collagen deposition and its remodelling in various other organs in health and disease.

【 授权许可】

CC BY