PROJECT SUMMARY: Organs and other complex tissues “know” when and how to stop growing to arrive at the correct size and shape. Disruption of organ size control mechanisms can lead to congenital abnormalities, poor organ homeostasis and tissue repair, and tumors. Adult zebrafish caudal fins, including their complex skeleton and other tissues, perfectly regenerate to their original size and shape regardless of the nature or position of the injury. Therefore, zebrafish fin regeneration provides a compelling and genetically tractable vertebrate model to interrogate organ size control mechanisms. Prevailing models for robust fin size regeneration speculate that fin cells maintain a multitude of “positional identities” that somehow instruct different degrees of outgrowth. We propose a distinct and straightforward model that neatly explains how fin size and shape is restored without invoking molecularly encoded positional information. A key cell population at the distal end of the regenerating fin that we term the “niche” produces Wnt signals that promote fin outgrowth by sustaining progenitor cells. We identify Dachsund transcription factors as novel niche markers and show that the niche uniquely forms from intra-‐‑ray mesenchyme that populates the inside of the cylindrical, differentially sized, and progressively tapered fin rays. We show that the niche, and therefore Wnt, steadily dissipates as regeneration unfolds; once exhausted, growth stops. As such, regenerated fin size is dictated by the amount of niche formed upon damage – which is simply dependent on the availability of intra-‐‑ray mesenchyme and hence bone width at the damage site. This “transpositional scaling” model suggests that macro-‐‑scale fin size and shape is determined by mesenchyme-‐‑niche state transitions and self-‐‑restoring bone geometry rather than unique positional identities of individual cells. We will explore this paradigm and uncover underlying cell and molecular mechanisms for size control during fin regeneration by three Specific Aims: 1. Define intra-‐‑ray mesenchyme / distal niche lineage cell states, transitions, and fates, 2. Determine signaling and transcriptional mechanisms maintaining niche state and function, and 3. Determine niche “countdown timer” mechanisms using longfint2 zebrafish – which we show have a broken timer due to misexpression of the kcnh2a ion channel. This insight suggests ion channels and Ca2+ signaling govern niche cell self-‐‑renewal. Our program will support a potentially broadly applicable “transpositional scaling” concept with exemplary mechanisms for how organ size and shape are determined by dynamic populations of tissue-‐‑resident niche cells....