Dataset Information

Toll-like receptor adaptor protein TIRAP has specialized roles in signaling, metabolic control and leukocyte migration upon wounding in zebrafish larvae

ABSTRACT: The Toll-interleukin-1 Receptor (TIR) domain-containing adaptor protein (TIRAP), also known as MyD88-associated ligand (MAL), is an essential adaptor protein in Toll-like receptor 2 (TLR2) signaling which links TLR2 and TLR4 to the other adaptor protein myeloid differentiation protein 88 (MyD88). Here, we investigate the transcriptomic and metabolomic profiles of zebrafish larvae from a tirap mutant and a myd88 mutant and the corresponding wild type controls under unchallenged normal developmental conditions. Comparing data from tirap and myd88 mutants with published data on tlr2 mutant, we found that Tirap has specialized roles in signaling and metabolic control. Transcriptome analysis revealed the specific involvement of tirap in calcium homeostasis and myosin regulation. Metabolomic profiling showed that the tirap mutation results in lower glucose levels, whereas a tlr2 mutation leads to higher glucose levels. Previously, tlr2 and myd88 have been indicated to be involved in responses to wounding by regulating leukocyte migration in zebrafish, but the role of tirap in regulating leukocyte migration has still not been investigated. In this study, a tail-wounding zebrafish larval model was used to identify the specific roles of tirap in leukocyte migration to tissue wounding. We found that more neutrophils were recruited to the wounded region in the tirap mutant larvae compared to the wild type controls, whereas there was no difference in macrophage recruitment. In contrast, published data show that tlr2 and myd88 mutants recruit fewer neutrophils and macrophages to the wounds. We visualized neutrophil migration trajectories in tail-wounding tirap mutant and wild type zebrafish larvae. Based on cell tracking analysis, we demonstrated that the neutrophil migration speed is increased in the tirap mutant. This is in contrast to our published results for neutrophil behavior in myd88 and tlr2 mutants. In conclusion, our findings demonstrate that tirap plays specialized roles distinct from tlr2 and myd88 in signaling, metabolic control, and in regulating neutrophil migration speed upon wounding.

ORGANISM(S): Danio rerio

PROVIDER: GSE270453 | GEO | 2025/01/01

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:MyD88 is an adaptor protein in Toll-like receptor and interleukin 1 receptor mediated signaling pathways that plays an essential role in activation of immune responses following pathogen recognition. We investigate that role in the zebrafish embryo model by using a zebrafish mutant line that contains a premature stop condon in the gene encoding MyD88, leading to a truncated protein that lacks domains important for its normal function. We infected these MyD88 mutants and wildtype individuals with Mycobacterium marinum to compare the resulting immune response by transcriptome profiling on total RNA isolated from single embryos. Autophagy regulator dram1 was identified as one of the MyD88-dependent genes. This RNAseq analysis was used to determine the effect of a truncation of the MyD88 protein on the innate immune response of zebrafish embryos during infection with Mycobacterium marinum. Myd88 mutant and wild type embryos were derived by incrossing homozygous myd88 mutant parents (allele hu3568, van der Vaart et al., 2013, Disease models & mechanisms 6, 841-854) or their wildtype siblings. RNA was isolated from pools of 20 embryos at 4 days post infection (4 dpi). The following treatment groups were used: homozygous mutants mock-injected with PBS/2%PVP 4 dpi, (2) wildtype siblings mock-injected with PBS/2%PVP 4dpi, (3) M. marinum-infected homozygous mutants 4dpi, (4) M. marinum-infected wildtype siblings 4dpi. Embryos were grown at 28.5M-bM-^@M-^S30M-BM-0C in egg water and manually dechorionated at 24 hours post fertilization (hpf). Subsequently, embryos were infected at 28 hpf by micro-injecting 200 colony forming units (CFU) of Mycobacterium marinum Mma20 bacteria into the caudal vein, or were mock-injected with buffer (PBS/2%PVP) as a control. After injections embryos were transferred into fresh egg water and incubated for 4 days at 28M-BM-0C. After the incubation period, single embryos were snap-frozen in liquid nitrogen and RNA was isolated for RNAseq analysis.

Dataset Information

Toll-like receptor adaptor protein TIRAP has specialized roles in signaling, metabolic control and leukocyte migration upon wounding in zebrafish larvae

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets