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Variability of oxygen isotopes in environmental water is recorded in tooth enamel, providing a record of seasonal change, dietary variability, and mobility. Physiology dampens this variability, however, as oxygen passes from environmental sources into blood and forming teeth. We showcase two methods of high resolution, 2-dimensional enamel sampling, and conduct modeling, to report why and how environmental oxygen isotope variability is reduced in animal bodies and teeth. First, using two modern experimental sheep, we introduce a sampling method, die-saw dicing, that provides high-resolution physical samples (n = 109 and 111 sample locations per tooth) for use in conventional stable isotope and molecular measurement protocols. Second, we use an ion microprobe to sample innermost enamel in an experimental sheep (n = 156 measurements), and in a Pleistocene orangutan (n = 176 measurements). Synchrotron and conventional μCT scans reveal innermost enamel thicknesses averaging 18 and 21 μm in width. Experimental data in sheep show that compared to drinking water, oxygen isotope variability in blood is reduced to 70–90 %; inner and innermost enamel retain between 36 and 48 % of likely drinking water stable isotope range, but this recovery declines to 28–34 % in outer enamel. 2D isotope sampling suggests that declines in isotopic variability, and shifted isotopic oscillations throughout enamel, result from the angle of secretory hydroxyapatite deposition and its overprinting by maturation. This overprinting occurs at all locations including innermost enamel, and is greatest in outer enamel. These findings confirm that all regions of enamel undergo maturation to varying degrees and confirm that inner and innermost enamel preserve more environmental variability than other regions. We further show how the resolution of isotope sampling — not only the spatial resolution within teeth, but also the temporal resolution of water in the environment — impacts our estimate of how much variation teeth recover from the environment. We suggest inverse methods, or multiplication by standard factors determined by ecology, taxon, and sampling strategy, to reconstruct the full scale of seasonal environmental variability. We advocate for combined inverse modeling and high-resolution sampling informed by the spatiotemporal pattern of enamel formation, and at the inner or innermost enamel when possible, to recover seasonal records from teeth. 

Abstract Many explanations for Eocene climate change focus on the Southern Ocean—where tectonics influenced oceanic gateways, ocean circulation reduced heat transport, and greenhouse gas declines prompted glaciation. To date, few studies focus on marine vertebrates at high latitudes to discern paleoecological and paleoenvironmental impacts of this climate transition. The Tertiary Eocene La Meseta (TELM) Formation has a rich fossil assemblage to characterize these impacts;Striatolamia macrota, an extinct (†) sand tiger shark, is abundant throughout the La Meseta Formation. Body size is often tracked to characterize and integrate across multiple ecological dimensions. †S. macrotabody size distributions indicate limited changes during TELMs 2–5 based on anterior tooth crown height (n = 450, mean = 19.6 ± 6.4 mm). Similarly, environmental conditions remained stable through this period based on δ¹⁸O_PO4values from tooth enameloid (n = 42; 21.5 ± 1.6‰), which corresponds to a mean temperature of 22.0 ± 4.0°C. Our preliminaryε_Nd(n = 4) results indicate an early Drake Passage opening with Pacific inputs during TELM 2–3 (45–43 Ma) based on single unit variation with an overall radiogenic trend. Two possible hypotheses to explain these observations are (1) †S. macrotamodified its migration behavior to ameliorate environmental changes related to the Drake Passage opening, or (2) the local climate change was small and gateway opening had little impact. While we cannot rule out an ecological explanation, a comparison with climate model results suggests that increased CO₂produces warm conditions that also parsimoniously explain the observations.