A unique characteristic of mammals is a vertebral column with anatomically distinct regions, but when and how this trait evolved remains unknown. We reconstructed vertebral regions and their morphological disparity in the extinct forerunners of mammals, the nonmammalian synapsids, to elucidate the evolution of mammalian axial differentiation. Mapping patterns of regionalization and disparity (heterogeneity) across amniotes reveals that both traits increased during synapsid evolution. However, the onset of regionalization predates increased heterogeneity. On the basis of inferred homology patterns, we propose a “pectoral-first” hypothesis for region acquisition, whereby evolutionary shifts in forelimb function in nonmammalian therapsids drove increasing vertebral modularity prior to differentiation of the vertebral column for specialized functions in mammals.

Mammals and their closest fossil relatives are unique among tetrapods in expressing a high degree of pectoral girdle and forelimb functional diversity associated with fully pelagic, cursorial, subterranean, volant, and other lifestyles. However, the earliest members of the mammalian stem lineage, the “pelycosaur”-grade synapsids, present a far more limited range of morphologies and inferred functions. The more crownward nonmammaliaform therapsids display novel forelimb morphologies that have been linked to expanded functional diversity, suggesting that the roots of this quintessentially mammalian phenotype can be traced to the pelycosaur–therapsid transition in the Permian period. We quantified morphological disparity of the humerus in pelycosaur-grade synapsids and therapsids using geometric morphometrics. We found that disparity begins to increase concurrently with the emergence of Therapsida, and that it continues to rise until the Permo-Triassic mass extinction. Further, therapsid exploration of new regions of morphospace is correlated with the evolution of novel ecomorphologies, some of which are characterized by changes to overall limb morphology. This evolutionary pattern confirms that nonmammaliaform therapsid forelimbs underwent ecomorphological diversification throughout the Permian, with functional elaboration initially being more strongly expressed in the proximal end of the humerus than the distal end. The role of the forelimbs in the functional diversification of therapsids foreshadows the deployment of forelimb morphofunctional diversity in the evolutionary radiation of mammals.

Mammals are the only living members of the larger clade Synapsida, which has a fossil record spanning 320 Ma. Despite the fact that much of the ecological diversity of mammals has been considered in the light of limb morphology, the ecological comparability of mammals to their fossil forerunners has not been critically assessed. Because of the wide use of limb morphology in testing ecomorphological hypothesis about extinct tetrapods, we sought: (i) to estimate when in synapsid history, modern mammals become analogues for predicting fossil ecologies; (ii) to document examples of ecomorphological convergence; and (iii) to compare the functional solutions of distinct synapsid radiations. We quantitatively compared the forelimb shapes of the multiple fossil synapsid radiations to a broad sample of extant Mammalia representing a variety of divergent locomotor ecologies. Our results indicate that each synapsid radiation explored different areas of morphospace and arrived at functional solutions that reflected their distinctive ancestral morphologies. This work counters the narrative of non-mammalian synapsid forelimb evolution as a linear progression towards more mammalian morphologies. Instead, a disparate array of early-evolving shapes subsequently contracted towards more mammal-like forms.

Endothermy ("warm-bloodedness") underpins the ecological dominance of mammals and birds in diverse environmental settings^1-3. However, it is unclear when this crucial feature emerged during mammalian evolutionary history, as most fossil evidence is ambiguous^4-25. Here, we show that new information on this key evolutionary transition can be obtained from the morphology of the endolymph-lled semicircular ducts of the inner ear that monitor head rotations and are essential for motor coordination, navigation, and spatial awareness^26-31. Increased body temperature during the ectotherm-endotherm transition of mammal ancestors would decrease endolymph viscosity, negatively impacting the biomechanics of the semicircular ducts^32,33, while simultaneously increasing activity levels^34,35 required improved performance^36. Speci c morphological changes to the membranous ducts and enclosing bony canals were, therefore, necessary to maintain optimal functionality. We track these morphological changes in 341 vertebrates, including 56 extinct synapsids, and show that canals with relatively thin cross-sections and small radii of curvature are indicative of mammalian endothermy. This inner ear morphotype evolved abruptly ~233 million years ago, during the Late Triassic, in Mammaliamorpha. Our conclusion differs from previous suggestions3-17, and we interpret most stem-mammals as ectotherms. Endothermy as a crucial physiological characteristic joins other distinctive mammalian features that arose during this period of climatic instability^37-39. *Ricardo Araújo and Romain David contributed equally to this work. Main TextEndotherms can maintain high and nearly constant body temperature through metabolic heat production, allowing them to optimize chemical reactions and sustain aerobic activity for long periods of time while remaining relatively independent from external conditions 1,2,40 . Compared to ectotherms, endotherms are more active, travel farther, and achieve higher locomotor speeds, all at the expense of higher energy costs 34,35 (Supplementary Data 1). Consequently, extant mammals and birds occupy a variety of ecological niches unrivalled by other vertebrates. Endothermy is a quintessentially mammalian feature, intimately related to other hallmarks such as lactation, sweat glands and fur 2,3 . However, its evolution remains one of the great unsolved mysteries of palaeontology 2,3 . Lines of evidence invoked to identify the emergence of mammalian endothermy rely mostly on skeletal anatomical features, but also on ichnological, histological and isotopic information, which have been used as correlates for aerobic