Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes.

Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes.
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DOI:
10.1371/journal.pone.0287519
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发表时间:
2023
期刊:
影响因子:
3.7
通讯作者:
--
中科院分区:
综合性期刊3区
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--
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大多数动物与细菌建立长期的共生关系,这对正常的宿主生理至关重要。夏威夷鱿鱼(Euprymna scolopes)与生物发光细菌费氏弧菌(Vibrio fischeri)之间形成的共生关系是研究促进动物-细菌共生的分子机制的重要模型系统。乌贼从未定植的卵中孵化,这导致了乌贼定植测定的发展,这种测定是基于将培养的V. fischeri细胞引入新孵化的幼乌贼中。最近的研究表明,菌株在如何建立共生关系方面往往表现出很大的差异。因此,我们试图开发一种简化和可重复的方案,允许研究人员确定适当的接种水平,并提供一个平台,在不同的实验室标准化检测。在我们的方案中,我们采用了一种通常用于评估病原体传染性的方法来量化费氏弧菌菌株的共生能力。由此产生的度量,共生剂量-50 (SD50),估计了特定费氏弧菌菌株建立发光共生所需的接种量水平。与其他方案相比,我们的方法所需的动物数量减少了2 - 5倍。此外,本文提出的功率分析表明,该方案可以检测到不同菌株之间SD50的变化高达3倍。
Most animals establish long-term symbiotic associations with bacteria that are critical for normal host physiology. The symbiosis that forms between the Hawaiian squid Euprymna scolopes and the bioluminescent bacterium Vibrio fischeri serves as an important model system for investigating the molecular mechanisms that promote animal-bacterial symbioses. E. scolopes hatch from their eggs uncolonized, which has led to the development of squid-colonization assays that are based on introducing culture-grown V. fischeri cells to freshly hatched juvenile squid. Recent studies have revealed that strains often exhibit large differences in how they establish symbiosis. Therefore, we sought to develop a simplified and reproducible protocol that permits researchers to determine appropriate inoculum levels and provides a platform to standardize the assay across different laboratories. In our protocol, we adapt a method commonly used for evaluating the infectivity of pathogens to quantify the symbiotic capacity of V. fischeri strains. The resulting metric, the symbiotic dose-50 (SD50), estimates the inoculum level that is necessary for a specific V. fischeri strain to establish a light-emitting symbiosis. Relative to other protocols, our method requires 2–5-fold fewer animals. Furthermore, the power analysis presented here suggests that the protocol can detect up to a 3-fold change in the SD50 between different strains.
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