Diverse spatiotemporal calcium signaling profiles regulate migratory behavior in corneal epithelial wound healing
Embargo Date
2027-09-09
OA Version
Citation
Abstract
Coordinated calcium signaling is essential for cellular processes including avascular wound healing. We hypothesize that this post-injury cellular signaling in the cornea is highly ordered, regulates cellular migratory behavior, and is dysregulated in diabetes. Using live cell imaging and computational analysis we discovered that calcium signaling events after injury to the corneal epithelium exhibit a distinct hierarchy and are driven by a subpopulation of high-signaling cells in both in vitro and ex vivo model systems that is attenuated in diabetes. Groups of cells near the wound engage in cyclical patterns of event initiation and propagation, and the spatiotemporal locations of high-signaling cells are revealed using topographical mapping of signal intensity. We used hierarchical clustering to assess conditions under which signaling phenotype diversity, and specifically the presence of high-signaling cells, arises and is attenuated. The purinergic receptor P2X7 is essential for the generation of conductor cells during the late wound response, and conductor cells were found to express significantly less P2X7 than lower-signaling neighbors. The receptor P2Y2 and the channel protein Pannexin-1 modulated the proportion of cells within the population that became conductors, but Connexin-43 had no effect. Long-term imaging was used to correlate signaling phenotype and motility and revealed that in non-diabetic cell culture models, high-signaling cells exhibit significantly less migratory behavior than lower-signaling neighbors. In non-diabetic ex vivo murine globes, wing and basal cells near the wound exhibit calcium signaling event propagation and hierarchy. However, there are significant aberrations in wing and basal cell populations in diabetic murine models. Our results indicate that post-injury calcium signaling in the corneal epithelium is highly ordered, that signal profile diversity is essential for efficient wound healing, and breakdown of this order is an early indicator of disease. This contrasts with previously held hypotheses in the field that post-injury calcium signaling was based only on environmental conditions and did not exhibit significant order.
Description
2024