Novel cell-based system to assay cell-cell fusion during myotube formation.

IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL
Mari Isobe, Yumika Suzuki, Hideshi Sugiura, Masahiro Shibata, Yuki Ohsaki, Satoshi Kametaka
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引用次数: 4

Abstract

A live assay tool has been established to uncover the precise molecular mechanisms underlying complex cell fusion events in myoblasts. The novel cell-based assay, HiMy (HiBiT-based myoblast fusion), utilizes a recently developed split-luciferase technology. The assay successfully detected cell fusion in differentiating C2C12 myoblast cultures. This allowed us to measure mixing of the cytoplasm, which occurred several hours after the initiation of C2C12 differentiation. Unlike what was reported earlier, the fusion was detected a few hours after the initiation of differentiation. Thus, this assay is sensitive enough to monitor fusion events before they become detectable using conventional methods. Furthermore, a panel of laboratory compounds, including a variety of inhibitors of cellular enzymes or activities, were assayed using the HiMy assay. Lovastatin, a cholesterol biogenesis inhibitor, decreased HiMy activity by approximately 50%. In contrast, mevalonolactone, a precursor for cholesterol synthesis, increased fusion activity. These results confirmed the previous finding that the amount of cellular cholesterol positively correlates with the rate of myoblast fusion during myogenesis. These results indicate that the novel cell fusion assay is a quick, accurate, and robust method to monitor intercellular fusion events.

新的基于细胞的系统来测定肌管形成过程中的细胞-细胞融合。
已经建立了一种实时检测工具,以揭示成肌细胞中复杂细胞融合事件的精确分子机制。基于hibit的成肌细胞融合(HiBiT-based myoblast fusion)是一种新型的细胞检测方法,它利用了最近开发的分裂荧光素酶技术。该实验成功地检测了C2C12成肌细胞分化培养中的细胞融合。这使我们能够测量细胞质的混合,这发生在C2C12分化开始后的几个小时。与之前报道的不同,融合是在分化开始几个小时后检测到的。因此,这种检测方法足够灵敏,可以在常规方法检测到聚变事件之前对其进行监测。此外,一组实验室化合物,包括各种细胞酶或活性抑制剂,使用HiMy测定法进行测定。洛伐他汀是一种胆固醇生物生成抑制剂,可使HiMy活性降低约50%。相反,甲维洛内酯,胆固醇合成的前体,增加了融合活性。这些结果证实了先前的发现,即细胞胆固醇的数量与成肌细胞融合率在肌肉形成过程中呈正相关。这些结果表明,新的细胞融合试验是一种快速、准确和稳健的方法来监测细胞间融合事件。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biomedical Research-tokyo
Biomedical Research-tokyo 医学-医学:研究与实验
CiteScore
2.40
自引率
0.00%
发文量
19
审稿时长
>12 weeks
期刊介绍: Biomedical Research is peer-reviewed International Research Journal . It was first launched in 1990 as a biannual English Journal and later became triannual. From 2008 it is published in Jan-Apr/ May-Aug/ Sep-Dec..
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