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Title page for ETD etd-09112017-142409

Type of Document Dissertation
Author Shannon, Erica Kristine
URN etd-09112017-142409
Title Dissecting Complex Mechanisms of Calcium Influx in a Simple Wound System
Degree PhD
Department Cell and Developmental Biology
Advisory Committee
Advisor Name Title
Irina Kaverina Committee Chair
Ian Macara Committee Member
Jeffrey Davidson Committee Member
M. Shane Hutson Committee Member
  • calcium
  • drosophila
  • wound healing
  • diffusion
  • laser ablation
  • cavitation
Date of Defense 2017-08-21
Availability unrestricted
In normal epithelial wound repair, cells across an epithelial sheet begin a coordinated process of re-epithelialization within minutes of wounding. These coordinated behaviors are driven by a calcium wave, a rise in cytosolic calcium expanding away from the wound in a wave-like fashion. The calcium wave is evolutionarily conserved and is the earliest detectable wound response. Understanding the mechanisms of calcium influx and propagation may reveal fundamental aspects of wound detection and of cell coordination.

We observed multiple, distinct mechanisms of calcium influx and propagation around reproducible wounds in the Drosophila notum. First, extracellular calcium flows directly into cells through micro-tears on the cell surface. We were able to assess the role of micro-tears in calcium dynamics by using pulsed laser ablation, a common wounding method that generates exaggerated micro-tears. Pulsed laser ablation creates a cavitation bubble, which forms and collapses within microseconds of ablation and damages the plasma membranes of cells tens of microns away from the wound. Once inside the cells, our model predicts calcium diffuses to neighboring cells via gap junctions. Next, we observed a larger, wound-induced calcium wave that is driven by an unknown extracellular signal. This signal activates a Gαq mediated signaling cascade and induces calcium release from intracellular ER stores.

Our simple, pulsed laser ablation wounding model recapitulates a complex damage profile and reveals multiple patterns of calcium influx and propagation around a single wound. For this reason, this model has the potential to unite previous, and seemingly contradictory, findings regarding calcium dynamics in the wound healing field.

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