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使用 FluoZin-3 和 fura-2 监测胰腺β细胞系中游离细胞内 Cd 的急性积累。

Using FluoZin-3 and fura-2 to monitor acute accumulation of free intracellular Cd in a pancreatic beta cell line.

机构信息

Departments of Anatomy, College of Graduate Studies, Midwestern University, Downers Grove, IL, 60515, USA.

AbbVie Inc., Headquarters 1 N. Waukegan Road, North Chicago, IL, 60064, USA.

出版信息

Biometals. 2019 Dec;32(6):951-964. doi: 10.1007/s10534-019-00226-z. Epub 2019 Nov 21.

DOI:10.1007/s10534-019-00226-z
PMID:31754889
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7446769/
Abstract

The understanding of cellular Cd accumulation and toxicity is hampered by a lack of fluorescent indicators selective for intracellular free Cd ([Cd]). In this study, we used depolarized MIN6 mouse pancreatic beta cells as a model for evaluating [Cd] detection with commercially available fluorescent probes, most of which have been traditionally used to visualize [Ca] and [Zn]. We trialed a panel of 12 probes including fura-2, FluoZin-3, Leadmium Green, Rhod-5N, indo-1, Fluo-5N, and others. We found that the [Zn] probe FluoZin-3 and the traditional [Ca] probe fura-2 responded most consistently and robustly to [Cd] accumulation mediated by voltage-gated calcium channels. While selective detection of [Cd] by fura-2 required the omission of Ca from extracellular buffers, FluoZin-3 responded to [Cd] similarly in the presence or absence of extracellular Ca. Furthermore, we showed that FluoZin-3 and fura-2 can be used together for simultaneous monitoring of [Ca] and [Cd] in the same cells. None of the other fluorophores tested were effective [Cd] detectors in this model.

摘要

细胞内 Cd 积累和毒性的理解受到缺乏对细胞内游离 Cd([Cd])具有选择性的荧光指示剂的阻碍。在这项研究中，我们使用去极化的 MIN6 小鼠胰腺β细胞作为模型，评估了商业上可用的荧光探针检测[Cd]的能力，其中大多数探针传统上用于可视化[Ca]和[Zn]。我们试用了一组 12 个探针，包括 fura-2、FluoZin-3、Leadmium Green、Rhod-5N、indo-1、Fluo-5N 等。我们发现，[Zn]探针 FluoZin-3 和传统的[Ca]探针 fura-2对电压门控钙通道介导的[Cd]积累反应最一致和最强烈。虽然 fura-2 对[Cd]的选择性检测需要从细胞外缓冲液中去除 Ca，但 FluoZin-3 在存在或不存在细胞外 Ca 的情况下对[Cd]的反应相似。此外，我们表明 FluoZin-3 和 fura-2 可用于同时监测同一细胞内的[Ca]和[Cd]。在该模型中，测试的其他荧光染料均不是有效的[Cd]检测探针。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b93/7446769/b753b49958c7/nihms-1619808-f0008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b93/7446769/b1a3dc7640b9/nihms-1619808-f0003.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b93/7446769/02ae42ef71a0/nihms-1619808-f0005.jpg
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