A method for mouse pancreatic islet isolation and intracellular cAMP determination.
A method for mouse pancreatic islet isolation and intracellular cAMP determination.
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DOI：
10.3791/50374
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发表时间：
2014-06-25
期刊：
影响因子：
--
通讯作者：
Kimple ME
中科院分区：
文献类型：
--
作者：
Neuman JC;Truchan NA;Joseph JW;Kimple ME
失控的血糖是糖尿病的一个标志，会促进神经病变、肾病和视网膜病变等疾病的发生。随着糖尿病的流行，无论是免疫介导的1型糖尿病还是肥胖相关的2型糖尿病，旨在描述糖尿病的病理生理和治疗机制的研究都是至关重要的。朗格汉斯人胰岛的β细胞负责在血糖浓度升高时适当地分泌胰岛素。除了葡萄糖和其他营养物质外，β细胞还受到特殊激素的刺激，称为胰岛素，这种激素是由肠道分泌的，作用于β细胞受体，增加细胞内环磷酸腺苷(CAMP)的产生。众所周知，β细胞功能、质量和胰岛素反应性降低与2型糖尿病的病理生理学有关，也越来越多地与1型糖尿病有关。目前的小鼠胰岛分离和cAMP测定方法可以帮助描述促进疾病进展和治疗干预的机制，特别是那些由胰岛素受体或相关受体介导的机制，这些受体通过调节细胞内cAMP的产生发挥作用。虽然将只描述cAMP测量，但所描述的胰岛分离方案创建了一种干净的制剂，还允许许多其他下游应用，包括葡萄糖刺激的胰岛素分泌、[~3H]-胸腺嘧啶核苷掺入、蛋白质丰度和mRNA表达。
Uncontrolled glycemia is a hallmark of diabetes mellitus and promotes morbidities like neuropathy, nephropathy, and retinopathy. With the increasing prevalence of diabetes, both immune-mediated type 1 and obesity-linked type 2, studies aimed at delineating diabetes pathophysiology and therapeutic mechanisms are of critical importance. The β-cells of the pancreatic islets of Langerhans are responsible for appropriately secreting insulin in response to elevated blood glucose concentrations. In addition to glucose and other nutrients, the β-cells are also stimulated by specific hormones, termed incretins, which are secreted from the gut in response to a meal and act on β-cell receptors that increase the production of intracellular cyclic adenosine monophosphate (cAMP). Decreased β-cell function, mass, and incretin responsiveness are well-understood to contribute to the pathophysiology of type 2 diabetes, and are also being increasingly linked with type 1 diabetes. The present mouse islet isolation and cAMP determination protocol can be a tool to help delineate mechanisms promoting disease progression and therapeutic interventions, particularly those that are mediated by the incretin receptors or related receptors that act through modulation of intracellular cAMP production. While only cAMP measurements will be described, the described islet isolation protocol creates a clean preparation that also allows for many other downstream applications, including glucose stimulated insulin secretion, [3H]-thymidine incorporation, protein abundance, and mRNA expression.
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影响因子：
7.7
作者：
DIONNE, KE;COLTON, CK;YARMUSH, ML
通讯作者：
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DOI：
10.3791/255
发表时间：
2007-01-01
期刊：
Journal of visualized experiments : JoVE
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影响因子：
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作者：
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