Microfluidic Devices For Determining Dynamics Of Islets of Langerhans

胰腺中数百万个胰岛同步激素分泌的机制 是未知的。罗珀实验室的长期目标是解码蜂窝通信， 了解正常的生物学功能和疾病进展。本建议的目的是确定 产生同步快速和超昼夜胰岛素和胰高血糖素振荡的机制， 小岛中心假设是，多种机制协同工作，产生快速和超日 荷尔蒙释放的波动执行这项工作的基本原理是， 葡萄糖调节激素分泌的动力学将导致治疗方法的设计， 与糖尿病和其他代谢疾病相关的并发症。在强有力的初步数据的指导下， 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 对胰岛素同步化的夹带，和2）鉴定胰高血糖素分泌和同步化动力学。下 第一个目标是，将同时使用两种诱导胰岛同步化的方法，一种是负胰岛素/葡萄糖 具有时间延迟的反馈回路，以及另一个，M3受体的脉动激活。为了实现这一目标，A 将使用液滴微流体技术开发用于胰岛素测量的高速方法，并将用于 测试一系列的时间延迟和并行灌注多种促分泌素的能力。第二个目标， 将首次使用均相免疫组织化学法测量单个胰岛的胰高血糖素分泌。 使用微流体系统的时间分辨荧光测定。通过这种方法，我们可以观察到单个 胰岛胰高血糖素分泌动力学，并将并行化的方法，以辨别胰高血糖素脉冲是如何 在多个小岛上同步。这项研究是创新的，因为微流体系统和 在这个提议中开发的测量方法将允许胰岛分泌的快速和超昼夜振荡 首次被观察到。这些结果将大大增加人们对胰岛功能的认识。 调节，这对于充分理解葡萄糖稳态机制及其如何出错至关重要 代谢性疾病。最终，这些知识有可能指导治疗发展，以减少 与II型糖尿病中血糖水平不受控制相关的问题。

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.