Reflex and habituation behavior of Caenorhabditis elegans assessed by a mechanical vibration system and image analysis

Yang, Wan-Hua; Chen, Chia-Yi; Wang, Kun-Liang; Kwok, Hong Luen; Stern, Arnold; Lo, Szecheng J.; Yang, Hung‐Chi

doi:10.1016/j.jneumeth.2019.108415

Cited by 1 publication

(1 citation statement)

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“…The assay was carried out with 60 worms at the L4 stage and scored for survival over time. In brief, synchronized L1 larvae were obtained from gravid adults by bleach based on a previous protocol [ 37 ]. The L1 larvae were cultured on E. coli HT115 until L4 stage before transferring to S. maltophilia -seeded NGM agar plates supplemented with 50 μM fluorodeoxyuridine (FUdR) to maintain a synchronous worm population [ 38 ].…”

Section: Methodsmentioning

confidence: 99%

PhoPQ two-component regulatory system plays a global regulatory role in antibiotic susceptibility, physiology, stress adaptation, and virulence in Stenotrophomonas maltophilia

Wu²,

Huang³

et al. 2020

BMC Microbiol

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Background Stenotrophomonas maltophilia, an opportunistic pathogen, is ubiquitously present in various environments, signifying its high capability of environmental adaptation. Two-component regulatory system (TCS) is a powerful implement to help organisms to survive in different environments. In clinic, treatment of S. maltophilia infection is difficult because it is naturally resistant to many antibiotics, highlighting the necessity to develop novel drugs or adjuvants. Given their critical and extensively regulatory role, TCS system has been proposed as a convincing target for novel drugs or adjuvants. PhoPQ TCS, a highly conserved TCS in several pathogens, plays crucial roles in low-magnesium adaption, polymyxin resistance, and virulence. In this study, we aimed to characterize the role of PhoPQ TCS of S. maltophilia in antibiotic susceptibility, physiology, stress adaptation, and virulence. Results To characterize PhoPQ system, phoP single mutant as well as phoP and phoQ double mutant were constructed. Distinct from most phoPQ systems of other microorganisms, two features were observed during the construction of phoP and phoQ single deletion mutant. Firstly, the phoQ mutant was not successfully obtained. Secondly, the compromised phenotypes of phoP mutant were not reverted by complementing an intact phoP gene, but were partially restored by complementing a phoPQ operon. Thus, wild-type KJ, phoP mutant (KJΔPhoP), phoPQ mutant (KJΔPhoPQ), and complemented strain (KJΔPhoPQ (pPhoPQ)) were used for functional assays, including antibiotic susceptibility, physiology (swimming motility and secreted protease activity), stress adaptation (oxidative, envelope, and iron-depletion stresses), and virulence to Caenorhabditis elegans. KJΔPhoPQ totally lost swimming motility, had enhanced secreted protease activity, increased susceptibility to antibiotics (β-lactam, quinolone, aminoglycoside, macrolide, chloramphenicol, and sulfamethoxazole/ trimethoprim), menadione, H2O2, SDS, and 2,2′-dipyridyl, as well as attenuated virulence to C. elegans. Trans-complementation of KJΔPhoPQ with phoPQ reverted these altered phenotypes to the wild-type levels. Conclusions Given the critical and global roles of PhoPQ TCS in antibiotic susceptibility, physiology, stress adaptation, and virulence, PhoPQ is a potential target for the design of drugs or adjuvants.

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Section: Methodsmentioning

confidence: 99%