CALCIUM REGULATION IN ORAL HEALTH

描述（由申请人提供）：钙是广泛生物功能的关键调节剂，也是牙釉质组成中的关键元素。在釉质形成的成熟阶段，随着釉质晶体在宽度和厚度上的扩大，Ca 2+需求增加。钙必须到达形成的釉质层，但这一过程是如何在成釉细胞中调节的知之甚少。钙离子在牙釉质中的转运模型主要关注钙离子通过进入、转运和排出的跨细胞通道。虽然有几个工作报告的运输和挤出步骤，只有有限的信息是可用于钙离子进入成釉细胞的机制。在这项资助计划中，我专注于通过检查钙库操作的钙释放激活的钙（CRAC）通道的作用的入口步骤。CRAC通道是上皮细胞中重要的Ca2+内流机制。除了免疫系统缺陷外，CRAC通道（STIM1，ORAI1）突变的患者还存在低钙化的釉质发生障碍。我的目标是确定Stim1和Orai1如何参与Ca2+进入以及如何调节这一过程。PI最近的工作表明，Stim1，Orai 1以及Ca2+信号胆囊收缩素（Cck）和Cck抑制剂Rcan 1被鉴定为在成熟过程中显著上调。我已经通过qPCR、Western blot和IHC证实了这些结果。因此，Ca2+通过CRAC通道流入成釉细胞以及如何调节这一过程对于健康釉质的发育至关重要。拟议拨款申请提供的证据将有助于更好地理解CRAC功能废除的系统性影响。这也将有助于医生做出关于牙医访问CRAC通道突变患者的决定，以防止牙齿问题 公共卫生相关性：钙对牙釉质的正常生物矿化至关重要。成釉细胞对钙的摄取受到干扰，导致成釉细胞的脱钙。我的目标是分析钙库操纵通道在釉质发育中的作用。

DESCRIPTION (provided by applicant): Calcium is a key regulator of a broad range of biological functions and is also a key element in the composition of dental enamel. During the maturation stage of amelogenesis, Ca2+ requirements increase as enamel crystals expand in width and thickness. Calcium must reach the forming enamel layer but how this process is regulated in ameloblasts is poorly understood. The current model for Ca2+ transport in enamel focuses on the transcellular passage of Ca2+ via the entry, transit and extrusion steps. Whereas several works have reported on the transit and extrusion steps, only limited information is available for Ca2+ entry mechanisms into ameloblasts. In this grant proposal, I focus on the entry step by examining the role of the store-operated Ca2+ release-activated Ca2+ (CRAC) channels. CRAC channels comprise important Ca2+ influx mechanism in epithelial cells. Patients with mutations to CRAC channels (STIM1, ORAI1) present, in addition to immune system deficiencies, with hypocalcified amelogenesis imperfecta. My goal is to identify how Stim1 and Orai1 are involved in Ca2+ entry and how this process is regulated. Recent work by the PI indicates that Stim1, Orai1 as well as well as the Ca2+ signaling cholecystokinin (Cck) and the Cck inhibitor Rcan1 were identified as being significantly up-regulated in maturation. I have confirmed these results by qPCR, Western blot and IHC. Thus Ca2+ influx into ameloblasts via CRAC channels and how this process is regulated is important for the development of healthy enamel. Evidence contributed by the proposed grant application will help to better understand the systemic effects of CRAC function abrogation. This will also help medical practitioners in making decisions concerning dentist visits to patients with mutations to CRAC channels in order to prevent dental problems PUBLIC HEALTH RELEVANCE: Calcium is critical for the normal biomineralization of enamel. Disruptions to calcium uptake by ameloblasts results in amelogenesis imperfecta. My goal is to analyze the function of calcium-store operated channels in enamel development.