Structural and functional polarisation of human pancreatic beta cells in islets from organ donors with and without type 2 diabetes.
Structural and functional polarisation of human pancreatic beta cells in islets from organ donors with and without type 2 diabetes.
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患有和不患有2型糖尿病的器官捐赠者胰岛中人胰腺β细胞的结构和功能极化。
DOI：
10.1007/s00125-020-05345-8
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发表时间：
2021-03
期刊：
影响因子：
8.2
通讯作者：
Thorn P
中科院分区：
文献类型：
--
作者：
Cottle L;Gan WJ;Gilroy I;Samra JS;Gill AJ;Loudovaris T;Thomas HE;Hawthorne WJ;Kebede MA;Thorn P
我们假设人类β细胞在胰岛毛细血管的结构和功能上是极化的。我们开始使用共聚焦显微镜绘制关键蛋白质的三维空间排列图和活细胞成像来确定胰岛素颗粒融合在细胞周围的分布。使用胰腺切片技术快速固定和处理人类胰腺样本，保持胰岛的结构和结构。用免疫荧光法对切片进行极性标记（scribble、disc large [Dlg]和分区缺陷3同源物[Par3]）和突触前标记（lipin、rab3相互作用蛋白[RIM2]和piccolo）染色，并使用3D共聚焦显微镜成像。分离的人胰岛分散培养在层粘连蛋白511包被的盖层上。用活体三维双光子显微镜观察培养细胞在葡萄糖刺激下的胞外颗粒融合事件。对人类胰岛内分泌细胞分布的评估发现，尽管不同的胰岛之间的复杂性和可变性不同，包括多小叶胰岛，以及α细胞和β细胞的混合，但在胰岛套膜处仍有显著的α细胞富集。对细胞位置的测量表明，大多数β细胞与胰岛毛细血管接触。在细胞下，β细胞一致地定位极性决定因子，如Par3、Dlg和scribble，基底结构域朝向毛细血管，顶结构域在对面。毛细血管界面/血管面富含突触前支架蛋白，如脂质、RIM2和短笛。有趣的是，突触前支架蛋白的富集也发生在β细胞接触胰岛周围毛细血管的地方，提示功能相互作用。我们还在2型糖尿病患者的胰岛中观察到相同的突触支架蛋白极化。与极化功能一致，分离的β细胞培养在层粘连蛋白包被的覆盖层上，目标胰岛素颗粒融合到覆盖层上。结构和功能极化是人类胰腺β细胞的一个决定性特征，在胰岛素分泌的控制中起着重要作用。在线版本包含同行评审但未经编辑的补充材料，可在10.1007/s00125-020-05345-8获得。
We hypothesised that human beta cells are structurally and functional polarised with respect to the islet capillaries. We set out to test this using confocal microscopy to map the 3D spatial arrangement of key proteins and live-cell imaging to determine the distribution of insulin granule fusion around the cells. Human pancreas samples were rapidly fixed and processed using the pancreatic slice technique, which maintains islet structure and architecture. Slices were stained using immunofluorescence for polarity markers (scribble, discs large [Dlg] and partitioning defective 3 homologue [Par3]) and presynaptic markers (liprin, Rab3-interacting protein [RIM2] and piccolo) and imaged using 3D confocal microscopy. Isolated human islets were dispersed and cultured on laminin-511-coated coverslips. Live 3D two-photon microscopy was used on cultured cells to image exocytic granule fusion events upon glucose stimulation. Assessment of the distribution of endocrine cells across human islets found that, despite distinct islet-to-islet complexity and variability, including multi-lobular islets, and intermixing of alpha and beta cells, there is still a striking enrichment of alpha cells at the islet mantle. Measures of cell position demonstrate that most beta cells contact islet capillaries. Subcellularly, beta cells consistently position polar determinants, such as Par3, Dlg and scribble, with a basal domain towards the capillaries and apical domain at the opposite face. The capillary interface/vascular face is enriched in presynaptic scaffold proteins, such as liprin, RIM2 and piccolo. Interestingly, enrichment of presynaptic scaffold proteins also occurs where the beta cells contact peri-islet capillaries, suggesting functional interactions. We also observed the same polarisation of synaptic scaffold proteins in islets from type 2 diabetic patients. Consistent with polarised function, isolated beta cells cultured onto laminin-coated coverslips target insulin granule fusion to the coverslip. Structural and functional polarisation is a defining feature of human pancreatic beta cells and plays an important role in the control of insulin secretion. The online version contains peer-reviewed but unedited supplementary material available at 10.1007/s00125-020-05345-8.
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3.2
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