BeschreibungRecent estimations indicate a global prevalence of 422 million people with diabetes in 2014. The two common types of diabetes (type 1 and type 2 diabetes) are on the one hand characterized by the autoimmune destruction of pancreatic β cells (type 1 diabetes) and on the other hand by reduced insulin secretion or insulin resistance (type 2 diabetes). Hence, there is an urgent need for new tools to study the molecular mechanism of insulin secretion and to identify substances which stimulate the release of insulin. Due to their low side effects, bioactive plant compounds and plant extracts are of special interest in this context. Existing methods such as patch-clamp measurements ,are very labor-intensive and difficult to handle. Here, we present the application of micro-patterned surfaces to measure the insulin release on a single cell level. To our knowledge there are no assays that base on this technology for the characterization of single β cells. For direct observation of fusion events of the secretory granules and the release of insulin, fluorescent assays in combination with micro-patterned substrates are an interesting approach. Glucose, as the physiological stimulus for insulin secretion in β cells is used to characterize and quantify the amount of secreted protein. For this reason, cells are grown on micro-patterned surfaces with specific capture antibodies against the protein of interest (insulin). A genetically encoded reporter called RINS1 was used for monitoring insulin secretion. Triggered by elevated glucose levels, secretory granules fuse with the plasma membrane and release their content that results in an increase in fluorescence monitored via total internal reflection fluorescence (TIRF) microscopy.
|Zeitraum||10 Okt. 2019|
|Ereignistitel||EMBO | EMBL Symposium|