Faculty Bibliography

Dental enamel is one of the most remarkable examples of matrix-mediated biomineralization. Enamel crystals form de novo in a rich extracellular environment in a stage-dependent manner producing complex microstructural patterns that are visually stunning. This process is orchestrated by specialized epithelial cells known as ameloblasts which themselves undergo striking morphological changes switching function from a secretory role to a cell primarily engaged in ionic transport. Ameloblasts are supported by a host of cell types which combined represent the enamel organ. Fully mineralized enamel is the hardest tissue found in vertebrates owing its properties partly to the unique mixture of ionic species represented and their highly organized assembly in the crystal lattice. Among the main elements found in enamel, Ca2+ is the most abundant ion yet how ameloblasts modulate Ca2+ dynamics remains poorly known. This review describes previously proposed models for passive and active Ca2+ transport, the intracellular Ca2+ buffering systems expressed in ameloblasts and provide an up-dated view of current models concerning Ca2+ influx and extrusion mechanisms, where most of the recent advances have been made. We also advance a new model for Ca2+ transport by the enamel organ

Loss-of-function mutations in stromal interaction molecule 1 (STIM1) impair the activation of Ca2+ release-activated Ca2+ (CRAC) channels and store-operated Ca2+ entry (SOCE), resulting in a disease syndrome called CRAC channelopathy that is characterized by severe dental enamel defects. The cause of these enamel defects has remained unclear given a lack of animal models. We generated Stim1/2K14cre mice to delete STIM1 and its homolog STIM2 in enamel cells. These mice showed impaired SOCE in enamel cells. Enamel in Stim1/2K14cre mice was hypomineralized with decreased Ca content, mechanically weak, and thinner. The morphology of SOCE-deficient ameloblasts was altered, showing loss of the typical ruffled border, resulting in mislocalized mitochondria. Global gene expression analysis of SOCE-deficient ameloblasts revealed strong dysregulation of several pathways. ER stress genes associated with the unfolded protein response were increased in Stim1/2-deficient cells, whereas the expression of components of the glutathione system were decreased. Consistent with increased oxidative stress, we found increased ROS production, decreased mitochondrial function, and abnormal mitochondrial morphology in ameloblasts of Stim1/2K14cre mice. Collectively, these data show that loss of SOCE in enamel cells has substantial detrimental effects on gene expression, cell function, and the mineralization of dental enamel.

Store-operated Ca2+ entry (SOCE) through Ca2+ release-activated Ca2+ (CRAC) channels is critical for lymphocyte function and immune responses. CRAC channels are hexamers of ORAI proteins that form the channel pore, but the contributions of individual ORAI homologues to CRAC channel function are not well understood. Here we show that deletion of Orai1 reduces, whereas deletion of Orai2 increases, SOCE in mouse T cells. These distinct effects are due to the ability of ORAI2 to form heteromeric channels with ORAI1 and to attenuate CRAC channel function. The combined deletion of Orai1 and Orai2 abolishes SOCE and strongly impairs T cell function. In vivo, Orai1/Orai2 double-deficient mice have impaired T cell-dependent antiviral immune responses, and are protected from T cell-mediated autoimmunity and alloimmunity in models of colitis and graft-versus-host disease. Our study demonstrates that ORAI1 and ORAI2 form heteromeric CRAC channels, in which ORAI2 fine-tunes the magnitude of SOCE to modulate immune responses.

The Malapa fossil assemblage was likely accumulated as a result of a death trap. Given this, the carnivoran species found there must have lived in proximity, close proximity for the smaller species, to the site, offering the possibility of expanding our interpretation of the habitats available to Australopithecus sediba via pinpoint palaeoenvironmental interpretation. To date, the identified carnivorans are the most abundant identified non-hominin taxa at Malapa, and given their territorial behaviour, are important when interpreting the palaeoecology of the site. The extinct false saber-tooth felid (Dinofelis barlowi) suggests that the presence of closed environments and the ancestral form of modern water mongoose (Atilax mesotes) indicates the presence of water in the vicinity. Canids generally support the presence of open habitats. The first appearance in the fossil record of Vulpes skinneri and Felis nigripes indicates the presence of drier open grassland/scrub. The Malapa carnivorans support widespread shifts in carnivore turnover circa 2.0 Ma in Africa and suggest, together with other lines of evidence, the occurrence of a regional transitioning environment during the time of Au. sediba.

Eccrine sweat glands are essential for sweating and thermoregulation in humans. Loss-of-function mutations in the Ca2+ release-activated Ca2+ (CRAC) channel genes ORAI1 and STIM1 abolish store-operated Ca2+ entry (SOCE), and patients with these CRAC channel mutations suffer from anhidrosis and hyperthermia at high ambient temperatures. Here we have shown that CRAC channel-deficient patients and mice with ectodermal tissue-specific deletion of Orai1 (Orai1K14Cre) or Stim1 and Stim2 (Stim1/2K14Cre) failed to sweat despite normal sweat gland development. SOCE was absent in agonist-stimulated sweat glands from Orai1K14Cre and Stim1/2K14Cre mice and human sweat gland cells lacking ORAI1 or STIM1 expression. In Orai1K14Cre mice, abolishment of SOCE was associated with impaired chloride secretion by primary murine sweat glands. In human sweat gland cells, SOCE mediated by ORAI1 was necessary for agonist-induced chloride secretion and activation of the Ca2+-activated chloride channel (CaCC) anoctamin 1 (ANO1, also known as TMEM16A). By contrast, expression of TMEM16A, the water channel aquaporin 5 (AQP5), and other regulators of sweat gland function was normal in the absence of SOCE. Our findings demonstrate that Ca2+ influx via store-operated CRAC channels is essential for CaCC activation, chloride secretion, and sweat production in humans and mice.

The purpose of this study was to measure the prevalence of enamel

The paradigm of chronobiology is based almost wholly upon the daily biological clock, or circadian rhythm, which has been the focus of intense molecular, cellular, pharmacological, and behavioral, research. However, the circadian rhythm does not explain biological timings related to fundamental aspects of life history such as rates of tissue/organ/body size development and control of the timing of life stages such as gestation length, age at maturity, and lifespan. This suggests that another biological timing mechanism is at work. Here we focus on a "many days" (multidien) chronobiological period first observed as enigmatic recurring growth lines in developing mammalian tooth enamel that is strongly associate with all adult tissue, organ, and body masses as well as life history attributes such as gestation length, age at maturity, weaning, and lifespan, particularly among the well studied primates. Yet, knowledge of the biological factors regulating the patterning of mammalian life, such as the development of body size and life history structure, does not exist. To identify underlying molecular mechanisms we performed metabolome and genome analyses from blood plasma in domestic pigs. We show that blood plasma metabolites and small non-coding RNA (sncRNA) drawn from 33 domestic pigs over a two-week period strongly oscillate on a 5-day multidien rhythm, as does the pig enamel rhythm. Metabolomics and genomics pathway analyses actually reveal two 5-day rhythms, one related to growth in which biological functions include cell proliferation, apoptosis, and transcription regulation/protein synthesis, and another 5-day rhythm related to degradative pathways that follows three days later. Our results provide experimental confirmation of a 5-day multidien rhythm in the domestic pig linking the periodic growth of enamel with oscillations of the metabolome and genome. This association reveals a new class of chronobiological rhythm and a snapshot of the biological bases that regulate mammalian growth, body size, and life history.

Haasgat is a primate-rich fossil locality in the northeastern part of the Fossil Hominid Sites of South Africa UNESCO World Heritage Site. Here we report the first hominin identified from Haasgat, a partial maxillary molar (HGT 500), that was recovered from an ex situ calcified sediment block sampled from the locality. The in situ fossil bearing deposits of the Haasgat paleokarstic deposits are estimated to date to slightly older than 1.95 Ma based on magnetobiostratigraphy. This places the hominin specimen at a critical time period in South Africa that marks the last occurrence of Australopithecus around 1.98 Ma and the first evidence of Paranthropus and Homo in the region between approximately 2.0 and 1.8 Ma. A comprehensive morphological evaluation of the Haasgat hominin molar was conducted against the current South African catalogue of hominin dental remains and imaging analyses using micro-CT, electron and confocal microscopy. The preserved occlusal morphology is most similar to Australopithecus africanus or early Homo specimens but different from Paranthropus. Occlusal linear enamel thickness measured from micro-CT scans provides an average of approximately 2.0 mm consistent with Australopithecus and early Homo. Analysis of the enamel microstructure suggests an estimated periodicity of 7-9 days. Hunter-Schreger bands appear long and straight as in some Paranthropus, but contrast with this genus in the short shape of the striae of Retzius. Taken together, these data suggests that the maxillary fragment recovered from Haasgat best fits within the Australopithecus-early Homo hypodigms to the exclusion of the genus Paranthropus. At approximately 1.95 Ma this specimen would either represent another example of late occurring Australopithecus or one of the earliest examples of Homo in the region. While the identification of this first hominin specimen from Haasgat is not unexpected given the composition of other South African penecontemporaneous site deposits, it represents one of the few hominin localities in the topographically-distinct northern World Heritage Site. When coupled with the substantial differences in the mammalian faunal communities between the northern localities (e.g., Haasgat, Gondolin) and well-sampled Bloubank Valley sites (e.g., Sterkfontein, Swartkrans, Kromdraai), the recovery of the HGT 500 specimen highlights the potential for further research at the Haasgat locality for understanding the distribution and interactions of hominin populations across the landscape, ecosystems and fossil mammalian communities of early Pleistocene South Africa. Such contextual data from sites like Haasgat is critical for understanding the transition in hominin representation at approximately 2 Ma sites in the region from Australopithecus to Paranthropus and early Homo.

Neanderthals had large and projecting (prognathic) faces similar to those of their putative ancestors from Sima de los Huesos (SH) and different from the retracted modern human face. When such differences arose during development and the morphogenetic modifications involved are unknown. We show that maxillary growth remodelling (bone formation and resorption) of the Devil's Tower (Gibraltar 2) and La Quina 18 Neanderthals and four SH hominins, all sub-adults, show extensive bone deposition, whereas in modern humans extensive osteoclastic bone resorption is found in the same regions. This morphogenetic difference is evident by approximately 5 years of age. Modern human faces are distinct from those of the Neanderthal and SH fossils in part because their postnatal growth processes differ markedly. The growth remodelling identified in these fossil hominins is shared with Australopithecus and early Homo but not with modern humans suggesting that the modern human face is developmentally derived.