Purification of pancreatic endocrine subsets reveals increased iron metabolism in beta cells
Purification of pancreatic endocrine subsets reveals increased iron metabolism in beta cells
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DOI：
10.1016/j.molmet.2020.101060
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发表时间：
2020-12-01
影响因子：
8.1
通讯作者：
Scharfmann, R.
中科院分区：
文献类型：
--
作者：
Berthault, C.;Staels, W.;Scharfmann, R.
目的：胰岛内分泌细胞主要有α、β和δ细胞。尽管这些细胞类型在葡萄糖稳态的调节中具有不同的作用，但不充分的纯化方法排除了对细胞类型特异性作用的研究。我们开发了一种可靠的方法，使活的α，β，和δ细胞从小鼠islets的下游analysis.Methods的同时分选：我们开发了一种抗体面板对细胞表面抗原，使高度纯化的内分泌亚群从小鼠胰岛的基础上的特定差异表达的CD71 β细胞和CD24 δ细胞的分离。我们严格证明了我们的方法的可靠性和有效性，使用散装和单细胞qPCR，免疫细胞化学，报告小鼠，和transcriptomics.Results：胰腺α，β和δ细胞可以分离的基础上β细胞特异性CD71表面表达和高表达的CD24 δ细胞。我们应用我们的新的分选策略来证明，CD71，这是转铁蛋白受体介导的转铁蛋白结合铁的摄取，是在β细胞在出生后早期周上调。我们发现，β细胞表达更高水平的其他几个基因参与铁代谢和铁剥夺显着损害β细胞功能。在人类β细胞，CD71同样需要铁的吸收和CD71的表面表达是受调控的葡萄糖依赖性mannes.Conclusions：这项研究提供了一种新的和有效的纯化方法，为鼠α，β和δ细胞，首次确定CD71作为出生后β细胞特异性标志物，并证明了铁代谢在β细胞功能的核心作用。(C)2020年，任作家。由爱思唯尔有限公司出版。
Objectives: The main endocrine cell types in pancreatic islets are alpha, beta, and delta cells. Although these cell types have distinct roles in the regulation of glucose homeostasis, inadequate purification methods preclude the study of cell type-specific effects. We developed a reliable approach that enables simultaneous sorting of live alpha, beta, and delta cells from mouse islets for downstream analyses.Methods: We developed an antibody panel against cell surface antigens to enable isolation of highly purified endocrine subsets from mouse islets based on the specific differential expression of CD71 on beta cells and CD24 on delta cells. We rigorously demonstrated the reliability and validity of our approach using bulk and single cell qPCR, immunocytochemistry, reporter mice, and transcriptomics.Results: Pancreatic alpha, beta, and delta cells can be separated based on beta cell-specific CD71 surface expression and high expression of CD24 on delta cells. We applied our new sorting strategy to demonstrate that CD71, which is the transferrin receptor mediating the uptake of transferrin-bound iron, is upregulated in beta cells during early postnatal weeks. We found that beta cells express higher levels of several other genes implicated in iron metabolism and iron deprivation significantly impaired beta cell function. In human beta cells, CD71 is similarly required for iron uptake and CD71 surface expression is regulated in a glucose-dependent manner.Conclusions: This study provides a novel and efficient purification method for murine alpha, beta, and delta cells, identifies for the first time CD71 as a postnatal beta cell-specific marker, and demonstrates a central role of iron metabolism in beta cell function. (C) 2020 The Authors. Published by Elsevier GmbH.