Faculty of Medicine, Institute of Physiology, University of Maribor, Maribor, Slovenia.
Faculty of Natural Sciences and Mathematics, University of Maribor, Maribor, Slovenia.
Am J Physiol Endocrinol Metab. 2021 Aug 1;321(2):E305-E323. doi: 10.1152/ajpendo.00043.2021. Epub 2021 Jul 19.
Many details of glucose-stimulated intracellular calcium changes in β cells during activation, activity, and deactivation, as well as their concentration-dependence, remain to be analyzed. Classical physiological experiments indicated that in islets, functional differences between individual cells are largely attenuated, but recent findings suggest considerable intercellular heterogeneity, with some cells possibly coordinating the collective responses. To address the above with an emphasis on heterogeneity and describing the relations between classical physiological and functional network properties, we performed functional multicellular calcium imaging in mouse pancreas tissue slices over a wide range of glucose concentrations. During activation, delays to activation of cells and any-cell-to-first-responder delays are shortened, and the sizes of simultaneously responding clusters increased with increasing glucose concentrations. Exactly the opposite characterized deactivation. The frequency of fast calcium oscillations during activity increased with increasing glucose up to 12 mM glucose concentration, beyond which oscillation duration became longer, resulting in a homogenous increase in active time. In terms of functional connectivity, islets progressed from a very segregated network to a single large functional unit with increasing glucose concentration. A comparison between classical physiological and network parameters revealed that the first-responders during activation had longer active times during plateau and the most active cells during the plateau tended to deactivate later. Cells with the most functional connections tended to activate sooner, have longer active times, and deactivate later. Our findings provide a common ground for recent differing views on β cell heterogeneity and an important baseline for future studies of stimulus-secretion and intercellular coupling. We assessed concentration-dependence in coupled β cells, degree of functional heterogeneity, and uncovered possible specialized subpopulations during the different phases of the response to glucose at the level of many individual cells. To this aim, we combined acute mouse pancreas tissue slices with functional multicellular calcium imaging over a wide range from threshold (7 mM) and physiological (8 and 9 mM) to supraphysiological (12 and 16 mM) glucose concentrations, classical physiological, and advanced network analyses.
许多关于β细胞在激活、活动和失活过程中葡萄糖刺激的细胞内钙离子变化的细节,以及它们的浓度依赖性,仍有待分析。经典的生理实验表明,在胰岛中,单个细胞之间的功能差异在很大程度上被减弱,但最近的发现表明存在相当大的细胞间异质性,其中一些细胞可能协调集体反应。为了强调异质性并描述经典生理学和功能网络特性之间的关系,我们在广泛的葡萄糖浓度范围内对小鼠胰腺组织切片进行了功能性多细胞钙成像。在激活过程中,细胞的激活延迟和任何细胞到第一个响应者的延迟缩短,同时响应的簇的大小随着葡萄糖浓度的增加而增加。相反,失活则完全相反。在活动期间,快速钙振荡的频率随着葡萄糖浓度的增加而增加,直到 12 mM 葡萄糖浓度,超过该浓度后,振荡持续时间变长,导致活跃时间的均匀增加。在功能连接方面,随着葡萄糖浓度的增加,胰岛从一个非常隔离的网络发展成一个单一的大型功能单元。经典生理学和网络参数的比较表明,在激活过程中,第一个响应者在平台期的活跃时间较长,而在平台期最活跃的细胞往往在后期失活。具有最多功能连接的细胞往往更早激活,具有更长的活跃时间,并且在后期失活。我们的发现为β细胞异质性的最近不同观点提供了一个共同点,为未来刺激-分泌和细胞间耦合的研究提供了一个重要的基线。我们评估了耦合β细胞的浓度依赖性、功能异质性程度,并在许多单个细胞水平上揭示了葡萄糖反应不同阶段可能存在的专门亚群。为此,我们将急性小鼠胰腺组织切片与广泛的功能多细胞钙成像相结合,范围从阈值(7 mM)和生理(8 和 9 mM)到超生理(12 和 16 mM)葡萄糖浓度,以及经典生理学和先进的网络分析。
Am J Physiol Endocrinol Metab. 2021-8-1
Methods Mol Biol. 2025
Front Endocrinol (Lausanne). 2022
Front Endocrinol (Lausanne). 2023
Acta Physiol (Oxf). 2025-2
Front Endocrinol (Lausanne). 2022
Elife. 2025-2-12
Histol Histopathol. 2025-9
Acta Physiol (Oxf). 2025-2
ArXiv. 2024-10-23
Methods Mol Biol. 2025