Integrated Spatiotemporal and Correlative Analysis of ATP-Related Signaling Components in Lens Development

Purpose: The development of the avascular lens depends on tightly regulated epithelial proliferation, differentiation into elongated fiber cells, and organelle elimination to maintain long-term transparency. Although Adenosine triphosphate (ATP) ectonucleotidases and Purinergic Receptors have been implicated in tissue morphogenesis, their specific contributions to lens development remain unclear. This study aimed to characterize the spatiotemporal expression patterns and predictive associations of ATP signaling components during lens development.

Methods: Transcriptomic datasets from human, mouse, and chicken lenses were analyzed to map the spatiotemporal expression of ATP ectonucleotidases and purinergic receptors. Pathway enrichment and correlation analyses with key developmental genes were performed to explore their functional associations. In vitro experiments were conducted using HLE‑B3 cells and primary embryonic chick lens cultures to assess cell viability, proliferation, Apoptosis, and differentiation under ATP manipulation and pharmacological P2 receptor blockade.

Results: Transcriptomic profiling revealed region- and stage-specific expression of ATP ectonucleotidases and purinergic receptors, which were functionally linked to calcium signaling, Autophagy, FGFR pathways, and glutathione-dependent redox regulation. In vitro, ATP exerted biphasic effects on HLE-B3 cells: low concentrations enhanced cell viability, whereas high concentrations increased epithelial-mesenchymal transition (EMT)-associated gene expression. In embryonic chick lens cultures, depletion of extracellular ATP or inhibition of P2 receptors reduced expression of key differentiation markers.

Conclusion: Integrated bioinformatic and experimental evidence indicate that ATP signaling components display distinct spatiotemporal expression patterns and possible associations with multiple functional pathways. In addition, various purinergic receptor subtypes may be implicated in coordinated and potentially diverse processes during lens development and the maintenance of transparency. Together, these correlative findings provide insights into the involvement of ATP and purinergic signaling in lens morphogenesis and homeostasis.

ATP; Lens development; ectonucleotidases; lens differentiation; purinergic signaling.