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

Josie Morris, Stephen Cross, Yinhui Lu, Karl E Kadler, Yongbo Lu, Sarah Dallas, Paul Martin*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

5 Citations (Scopus)
183 Downloads (Pure)

Abstract

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.

Original languageEnglish
Pages (from-to)4-11
Number of pages8
JournalDevelopmental Biology
Volume441
Issue number1
Early online date6 Jun 2018
DOIs
Publication statusPublished - 1 Sep 2018

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Keywords

  • Collagen-I
  • Live imaging
  • Skin
  • Wound healing
  • Zebrafish larvae

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