Calcium Regulation and Oral Health

项目概要/摘要 钙是一个广泛的生物功能的关键调节剂，也是组成中的关键元素 牙釉质在釉质形成的成熟阶段，随着釉质的发育， 晶体在宽度和厚度上膨胀。钙必须达到形成釉质层，但这个过程是如何 在成釉细胞中的调控机制知之甚少。目前釉质中Ca 2+转运的模型集中在 Ca 2+通过进入、转运和挤出步骤跨细胞通过。尽管有几个作品报道说 在运输和挤出步骤中，只有有限的信息可用于Ca 2+进入机制， 造釉细胞在这个资助计划中，我通过研究钙库运作的钙离子的作用来关注进入步骤。 释放激活的Ca 2+（CRAC）通道。CRAC通道包括重要的Ca 2+内流机制， 上皮细胞除了免疫外，还存在CRAC通道（STIM 1、ORR 1）突变的患者 系统缺陷，伴有低钙化的釉质形成障碍。我的目标是确定Stim 1和Orai 1 参与Ca 2+进入以及如何调节这一过程。PI最近的工作表明，Stim 1，Orai 1 以及Ca 2+信号胆囊收缩素（Cck）和Cck抑制剂Rcan 1被鉴定为是 在成熟期显著上调。我已经通过qPCR、Western blot和IHC证实了这些结果。因此 Ca 2+通过CRAC通道流入成釉细胞以及该过程如何调节对于成釉细胞的增殖和分化是重要的。 发展健康的牙釉质。拟议的赠款申请提供的证据将有助于更好地 了解CRAC功能消除的全身效应。这也将有助于医生在 做出关于牙医访问具有CRAC通道突变的患者的决定，以防止 牙齿问题

PROJECT SUMMARY/ABSTRACT 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