Microfluidic Devices to Determine Roles of Islet-Secreted Leptin

描述（由申请人提供）：瘦素由胰岛产生和分泌，但瘦素在这种情况下的分泌动力学和功能尚不清楚。长期目标是定量和同时监测参与脂肪细胞和胰岛之间通讯途径的肽和蛋白质的分泌。本申请的总体目标是开发技术以测量来自胰岛的瘦素分泌的急性变化并确定这种分泌如何影响传统肽的释放。核心假设是胰岛分泌的瘦素通过改变胰岛生理学来影响全身葡萄糖稳态。为了验证这一假设，将实现以下目标：1。胰岛分泌的瘦素的急性分泌动力学将在微流体装置上使用免疫PCR测定来确定，2.）瘦素对胰岛生理学的影响将通过使用多色电泳免疫测定在微流体装置上同时监测胰岛分泌的胰岛素、胰高血糖素和胰淀素来确定，和3）培养条件对胰岛分泌的瘦素的影响将使用在目标1和2中开发的技术的组合来确定。这项工作是重要的，因为胰岛分泌的瘦素的作用还没有被定义，并可能确定一个新的机制，影响血糖水平。开发的方法也将适用于其他生物系统的测量同时肽或蛋白质分泌。胰岛分泌的瘦素的功能尚未研究，但有可能影响人类健康的广泛领域。拟议的研究与公共卫生有关，因为这些知识将增加我们对胰岛生物学的理解，并可能为减轻糖尿病和肥胖相关并发症提供新的途径。

DESCRIPTION (provided by applicant): Leptin is produced and secreted from islets of Langerhans, but the secretory dynamics and functions of leptin in this setting are unknown. The long-term goal is to quantitatively and simultaneously monitor secretion of peptides and proteins involved in the communication pathway between adipocytes and islets of Langerhans. The overall objective of this application is the development of technology to measure acute changes in leptin secretion from islets of Langerhans and determine how this secretion affects the release of traditional peptides. The central hypothesis is that islet-secreted leptin affects systemic glucose homeostasis by altering islet physiology. To test this hypothesis, the following aims will be accomplished: 1.) acute secretory dynamics of islet-secreted leptin will be determined using an immuno-PCR assay on a microfluidic device, 2.) the effect of leptin on islet physiology will be ascertained by simultaneously monitoring insulin, glucagon, and amylin secretion from islets of Langerhans on a microfluidic device using a multi-color electrophoretic immunoassay, and 3.) the effects that culture conditions have on islet-secreted leptin will be determined using a combination of the technology developed in aims 1 and 2. This work is significant as the role of islet-secreted leptin has not been defined and may identify a new mechanism that influences blood glucose levels. The methodology developed will also be applicable to other biological systems for measurement of simultaneous peptide or protein secretion. The function of islet-secreted leptin has not been investigated, but has the potential to impact broad areas of human health. The proposed research has relevance to public health because this knowledge will increase our understanding of islet biology and potentially provide new routes for alleviating complications associated with diabetes and obesity.