AUTHOR=Hu Wanbin , van Steijn Leonie , Li Chen , Verbeek Fons J. , Cao Lu , Merks Roeland M. H. , Spaink Herman P. 

TITLE=A Novel Function of TLR2 and MyD88 in the Regulation of Leukocyte Cell Migration Behavior During Wounding in Zebrafish Larvae

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=9

YEAR=2021

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2021.624571

DOI=10.3389/fcell.2021.624571

ISSN=2296-634X

ABSTRACT=<p>Toll-like receptor (TLR) signaling via myeloid differentiation factor 88 protein (MyD88) has been indicated to be involved in the response to wounding. It remains unknown whether the putative role of MyD88 in wounding responses is due to a control of leukocyte cell migration. The aim of this study was to explore <italic>in vivo</italic> whether TLR2 and MyD88 are involved in modulating neutrophil and macrophage cell migration behavior upon zebrafish larval tail wounding. Live cell imaging of tail-wounded larvae was performed in <italic>tlr2</italic> and <italic>myd88</italic> mutants and their corresponding wild type siblings. In order to visualize cell migration following tissue damage, we constructed double transgenic lines with fluorescent markers for macrophages and neutrophils in all mutant and sibling zebrafish lines. Three days post fertilization (dpf), tail-wounded larvae were studied using confocal laser scanning microscopy (CLSM) to quantify the number of recruited cells at the wounding area. We found that in both <italic>tlr2<sup>–/–</sup></italic> and <italic>myd88<sup>–/–</sup></italic> groups the recruited neutrophil and macrophage numbers are decreased compared to their wild type sibling controls. Through analyses of neutrophil and macrophage migration patterns, we demonstrated that both <italic>tlr2</italic> and <italic>myd88</italic> control the migration direction of distant neutrophils upon wounding. Furthermore, in both the <italic>tlr2</italic> and the <italic>myd88</italic> mutants, macrophages migrated more slowly toward the wound edge. Taken together, our findings show that <italic>tlr2</italic> and <italic>myd88</italic> are involved in responses to tail wounding by regulating the behavior and speed of leukocyte migration <italic>in vivo</italic>.</p>