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Abstract

The central functional unit of the vertebrate eye is the retina, composed of neural retina (NR), retinal pigmented epithelium (RPE), and non-visual retina (NVR). In amphibians and fish, the retina grows throughout life via different pools of stem cells (SCs). In this work, I combined experimental and computational approaches to elucidate SC dynamics in the three retinal tissues of the teleost fish medaka (Oryzias latipes). I developed a cell centred agent based model to recapitulate post-embryonic growth of the NR and RPE. By accounting for 3D tissue geometry and continuous growth, the model reconciled conflicting hypotheses, demonstrating that competition between SCs is not mutually exclusive with lifelong coexistence of multiple SC lineages. To understand how NR and RPE regulate their proliferative output to coordinate growth rates, I developed quantitative methods to compare experiment and simulation. I tested the experimental data against simulations implementing two modes of feedback between cell proliferation and organ growth. Thus, I identified that the NR acts upstream to set the growth pace by sending an inductive growth signal, while the RPE responds downstream to this signal. Leveraging the model, I showed that NR SCs compete for niche space, but tissue geometry biases cells at certain positions to win this competition. Further, NR SCs modulate division axes and proliferation rate to change organ shape and retinal topology. Motivated by model predictions, I experimentally characterised the large SC population of the RPE, which consisted of both cycling and non-cycling quiescent cells. Putative sister cells exhibited similar temporal dynamics in local clusters, indicating that quiescence was the major mechanism for regulating proliferative output in the RPE. Finally, I experimentally showed that the NVR grows post-embryonically from a primordium, and shared all known markers for NR SCs in the same spatial distribution. Unlike NR and RPE, the NVR lacked a dedicated niche, instead proliferative cells were distributed throughout the tissue. Lineage tracing revealed a continuous relationship between RPE, NVR, and NR. Thus, the SCs of NR and RPE, and all cells of the NVR displayed plastic multipotency capable of generating all retinal tissues. By taking advantage of the positive feedback loop between experiment and simulation, this work shines a new light into a fundamental problem – growth coordination of different SC populations in a complex vertebrate organ.