NEST Laboratory-Scuola Normale Superiore, Piazza San Silvestro 12, 56127 Pisa, Italy.
Islet Cell Laboratory, Department of Clinical and Experimental Medicine, University of Pisa, 56126 Pisa, Italy.
Int J Mol Sci. 2024 Sep 27;25(19):10391. doi: 10.3390/ijms251910391.
Pro-inflammatory cytokines play a role in the failure of β cells in type 1 and type 2 diabetes. While existing data from 'omics' experiments allow for some understanding of the molecular mechanisms behind cytokine-induced dysfunction in β cells, no report thus far has provided information on the direct imaging of the β cell landscape with nanoscale resolution following cytokine exposure. In this study, we use Airyscan-based optical super-resolution microscopy of Insulinoma 1E (INS-1E) cells to investigate the structural properties of two subcellular membranous compartments involved in the production, maturation and secretion of insulin-containing granules, the endoplasmic reticulum (ER) and the Golgi apparatus (GA). Our findings reveal that exposure of INS-1E cells to IL-1β and IFN-γ for 24 h leads to significant structural alterations of both compartments. In more detail, both the ER and the GA fragment and give rise to vesicle-like structures with markedly reduced characteristic area and perimeter and increased circularity with respect to the original structures. These findings complement the molecular data collected thus far on these compartments and their role in β cell dysfunction and lay the groundwork for future optical microscopy-based ex vivo and in vivo investigations.
促炎细胞因子在 1 型和 2 型糖尿病中β细胞衰竭中起作用。虽然来自“组学”实验的现有数据允许对细胞因子诱导的β细胞功能障碍背后的分子机制有一定的了解,但迄今为止,没有报告提供关于细胞因子暴露后β细胞景观的纳米级分辨率直接成像的信息。在这项研究中,我们使用基于 Airyscan 的光学超分辨率显微镜研究胰岛素瘤 1E (INS-1E) 细胞,以研究与胰岛素分泌颗粒的产生、成熟和分泌有关的两个亚细胞膜结构的结构特性,内质网 (ER) 和高尔基体 (GA)。我们的研究结果表明,IL-1β和 IFN-γ 暴露 INS-1E 细胞 24 小时会导致这两个隔室的结构发生显著变化。更详细地说,内质网和高尔基体都分裂,并产生具有明显减小的特征面积和周长以及增加的圆度的囊泡样结构,与原始结构相比。这些发现补充了迄今为止对这些隔室及其在β细胞功能障碍中的作用的分子数据,并为未来基于光学显微镜的离体和体内研究奠定了基础。