Microfluidic Devices For Determining Dynamics Of Islets of Langerhans

胰腺内数百万个朗格汉斯胰岛间激素分泌同步的机制 都是未知的。罗珀实验室的长期目标是解码蜂窝通信，以使 了解正常的生物功能和疾病进展。这项提案的目标是确定 胰岛素和胰高血糖素产生同步、快速和超常振荡的机制 小岛。中心假设是，多种机制协同工作，既能产生快速效应，又能产生超常效应。 荷尔蒙释放的振荡。执行这项工作的基本原理是，对 葡萄糖调节激素分泌的动态将导致治疗方法的设计，以减轻 与糖尿病和其他代谢性疾病相关的并发症。在强劲的初步数据的指引下， 这一假设将通过追求两个具体目标来检验：1)确定时间延迟和双重时间的影响 夹带对胰岛素同步化的影响，以及2)确定胰升糖素的分泌和同步化动力学。在……下面 第一个目标是，两种诱导胰岛同步化的方法将一起使用，一种是负胰岛素/葡萄糖 反馈环有时间延迟，另一个是M3受体的脉动性激活。为了实现这一目标，一个 将利用液滴微流控技术开发高速胰岛素测量方法，并将用于 测试一系列时间延迟和并行灌流多个分泌器的能力。在第二个目标中， 将首次使用同种异体胰岛测量朗格汉斯单个胰岛的胰升糖素分泌 利用微流控系统进行时间分辨荧光分析。使用这种方法，我们预计将观察到单个 胰岛高血糖素分泌动力学并将并行化的方法来辨别高血糖素的脉搏如何 在多个小岛上同步。拟议的研究具有创新性，因为微流控系统和 在这项建议中开发的测量方法将允许胰岛分泌的快速和超传统振荡。 第一次被观察到。这些结果将大大增加人们对胰岛的认识 调节，这对于充分理解葡萄糖稳态的机制和它是如何出错的至关重要 代谢性疾病。最终，这些知识有可能指导治疗方法的发展，以减少 与II型糖尿病患者血糖水平不受调节有关的问题。

The mechanisms that synchronize hormone secretion across millions of islets of Langerhans in the pancreas are unknown. The long-term goal of the Roper laboratory is to decode cellular communication to enable understanding of normal biological function and disease progression. The objective of this proposal is to identify the mechanisms that generate synchronized rapid and ultradian insulin and glucagon oscillations from multiple islets. The central hypothesis is that multiple mechanisms working in concert produce both rapid and ultradian oscillations of hormone release. The rationale for performing this work is that a thorough understanding of the dynamics of glucose-regulatory hormone secretion will lead to the design of therapeutic approaches that alleviate the complications associated with diabetes and other metabolic diseases. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Determine the effect of time delays and dual entrainment on insulin synchronization, and 2) identify glucagon secretion and synchronization dynamics. Under the first aim, two methods for inducing islet synchronization will be used together, one a negative insulin/glucose feedback loop with time delays, and the other, pulsatile activation of M3 receptors. To accomplish this aim, a high-speed method for insulin measurement will be developed using droplet microfluidics and will be used for testing a range of time delays and the ability to perfuse multiple secretagogues in parallel. In the second aim, glucagon secretion will be measured for the first time from single islets of Langerhans using a homogeneous time resolved fluorescence assay using a microfluidic system. With this method, we anticipate observing single islet glucagon secretion dynamics and will parallelize the method to discern how glucagon pulses are synchronized across multiple islets. The proposed research is innovative because the microfluidic systems and measurement approaches developed in this proposal will allow rapid and ultradian oscillations of islet secretion to be observed for the first time. These results will provide a significant increase in the knowledge of islet regulation, which is crucial for fully understanding the mechanism of glucose homeostasis and how it goes awry in metabolic diseases. Ultimately, this knowledge has the potential to guide therapeutic development for reducing the problems associated with unregulated glucose levels in type II diabetes.