Microfluidic Devices to Determine Roles of Islet-Secreted Leptin

描述(申请人提供)：瘦素是由朗格汉斯的胰岛产生和分泌的，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与脂肪细胞和胰岛之间的通讯途径有关的多肽和蛋白质的分泌。这项应用的总体目标是开发技术来测量朗格汉斯胰岛瘦素分泌的急性变化，并确定这种分泌如何影响传统多肽的释放。中心假说是胰岛分泌的瘦素通过改变胰岛的生理功能来影响全身的血糖稳态。为了验证这一假设，将实现以下目标：1)胰岛分泌瘦素的急性分泌动力学将使用微流控设备上的免疫-聚合酶链式反应分析来确定，2。瘦素对胰岛生理的影响将通过在微流控装置上用多色电泳法同时监测胰岛胰岛素、胰升糖素和胰淀素的分泌来确定。培养条件对胰岛分泌瘦素的影响将使用AIMS 1和AIMS 2中开发的技术相结合来确定。这项工作意义重大，因为胰岛分泌瘦素的作用尚未确定，并可能确定影响血糖水平的新机制。所开发的方法也将适用于其他生物系统，以测量同时分泌的多肽或蛋白质。胰岛分泌的瘦素的功能尚未被研究，但有可能影响人类健康的广泛领域。这项拟议的研究与公众健康相关，因为这些知识将增加我们对胰岛生物学的理解，并可能为减轻与糖尿病和肥胖症相关的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.