高浓度荧光生物测定研究毒液诱导的孔隙形成、离子通道调节和细胞膜裂解

IF 3.6 Q2 TOXICOLOGY
Simon Kramer , Charan Kotapati , Yuanzhao Cao , Bryan G. Fry , Nathan J. Palpant , Glenn F. King , Fernanda C. Cardoso
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引用次数: 0

摘要

毒液由高度复杂的生物活性分子组成,可调节离子通道、受体、凝血因子和细胞膜。这一系列靶点和生物活性需要先进的高含量生物测定,以促进新型毒液疗法以及生物技术和药物制剂的开发。针对毒液研究方面的现有差距,我们开发了一种基于荧光的尖端高通量、高含量细胞检测方法。这种检测方法能同时鉴定毒液诱导的普遍细胞活动,如膜裂解、孔隙形成和离子通道调节。通过将细胞内钙测定与细胞外核酸测定相结合,我们成功地在单个细胞测定中区分了这些毒液机制。我们的高含量生物测定适用于三种细胞类型,它们分别暴露于代表溶解毒素、离子孔形成毒素或离子通道调节毒素的毒液成分。除了揭示这些作用机制的不同特征外,我们还发现孔形成潜伏毒素α-Lt1a更喜欢人类神经母细胞瘤而不是肾细胞和心肌细胞,而溶解蜂肽melittin则没有选择性。此外,对蛇毒的评估显示,蛇毒能诱导细胞膜快速裂解,而蝰蛇毒则对神经母细胞瘤细胞表现出不同程度的活性,甚至没有活性。这些发现凸显了我们的高含量生物测定在区分各支系和种间性状方面的能力,与毒液水平的临床观察结果一致,而不仅仅是区分离子孔形成、膜裂解和离子通道调节。我们希望我们的研究将加快对毒液生物学以及引起细胞毒性、心脏毒性和神经毒性效应的毒素多样性的理解,并帮助确定有可能造福人类的毒液成分。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

High-content fluorescence bioassay investigates pore formation, ion channel modulation and cell membrane lysis induced by venoms

High-content fluorescence bioassay investigates pore formation, ion channel modulation and cell membrane lysis induced by venoms

Venoms comprise highly sophisticated bioactive molecules modulating ion channels, receptors, coagulation factors, and the cellular membranes. This array of targets and bioactivities requires advanced high-content bioassays to facilitate the development of novel envenomation treatments and biotechnological and pharmacological agents. In response to the existing gap in venom research, we developed a cutting-edge fluorescence-based high-throughput and high-content cellular assay. This assay enables the simultaneous identification of prevalent cellular activities induced by venoms such as membrane lysis, pore formation, and ion channel modulation. By integrating intracellular calcium with extracellular nucleic acid measurements, we have successfully distinguished these venom mechanisms within a single cellular assay. Our high-content bioassay was applied across three cell types exposed to venom components representing lytic, ion pore-forming or ion channel modulator toxins. Beyond unveiling distinct profiles for these action mechanisms, we found that the pore-forming latrotoxin α-Lt1a prefers human neuroblastoma to kidney cells and cardiomyocytes, while the lytic bee peptide melittin is not selective. Furthermore, evaluation of snake venoms showed that Elapid species induced rapid membrane lysis, while Viper species showed variable to no activity on neuroblastoma cells. These findings underscore the ability of our high-content bioassay to discriminate between clades and interspecific traits, aligning with clinical observations at venom level, beyond discriminating among ion pore-forming, membrane lysis and ion channel modulation. We hope our research will expedite the comprehension of venom biology and the diversity of toxins that elicit cytotoxic, cardiotoxic and neurotoxic effects, and assist in identifying venom components that hold the potential to benefit humankind.

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来源期刊
Toxicon: X
Toxicon: X Pharmacology, Toxicology and Pharmaceutics-Toxicology
CiteScore
6.50
自引率
0.00%
发文量
33
审稿时长
14 weeks
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