DOI: 10.1101/456541Mar 20, 2019Paper

A transgenic zebrafish line for in vivo visualisation of neutrophil myeloperoxidase

BioRxiv : the Preprint Server for Biology
K. D. BuchanStephen A Renshaw

Abstract

The neutrophil enzyme myeloperoxidase (MPO) is a major enzyme made by neutrophils to generate antimicrobial and immunomodulatory compounds, notably hypochlorous acid (HOCl), amplifying their capacity for destroying pathogens and regulating inflammation. Despite its roles in innate immunity, the importance of MPO in preventing infection is unclear, as individuals with MPO deficiency are asymptomatic with the exception of an increased risk of candidiasis. Dysregulation of MPO activity is also linked with inflammatory conditions such as atherosclerosis, emphasising a need to understand the roles of the enzyme in greater detail. Consequently, new tools for investigating granular dynamics in vivo can provide useful insights into how MPO localises within neutrophils, aiding understanding of its role in preventing and exacerbating disease. The zebrafish is a powerful model for investigating the immune system in vivo, as it is genetically tractable, and optically transparent. To visualise MPO activity within zebrafish neutrophils, we created a genetic construct that expresses human MPO as a fusion protein with a C-terminal fluorescent tag, driven by the neutrophil-specific promoter lyz. After introducing the construct into the zebrafi...Continue Reading

Methods Mentioned

BETA
genotyping
transgenic
glycosylation
restriction digest
PCR
restriction

Software Mentioned

GraphPad
NIS Elements
Prism
Zeiss®

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