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Oosterhof, N.

Publications and source records attributed to Oosterhof, N..

2 recordsLinked to original sources

Zebrafish macrophage developmental arrest underlies depletion of microglia and reveals Csf1r-independent metaphocytes

Macrophages derive from multiple sources of hematopoietic progenitors. Most macrophages require colony-stimulating factor 1 receptor (CSF1R), but some macrophages persist in the absence of CSF1R. Here, we analyzed mpeg1:GFP-expressing macrophages in csf1r-deficient zebrafish and report that embryonic macrophages emerge followed by their developmental arrest. In larvae, mpeg1+ cell numbers then increased showing two distinct types in the skin: branched, putative Langerhans cells, and amoeboid cells. In contrast, although numbers also increased in csf1r-mutants, exclusively amoeboid mpeg1+ cells were present, which we showed by genetic lineage tracing to have a non-hematopoietic origin. They expressed macrophage-associated genes, but also showed decreased phagocytic gene expression and increased epithelial-associated gene expression, characteristic of metaphocytes, recently discovered ectoderm-derived cells. We further demonstrated that juvenile csf1r-deficient zebrafish exhibit systemic macrophage depletion. Thus, Csf1r deficiency disrupts embryonic to adult macrophage development. Csf1r-deficient zebrafish are viable and permit analyzing the consequences of macrophage loss throughout life.

developmental biology↗

Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brain

Microglia are brain resident macrophages, which have specialized functions important in brain development and in disease. They colonize the brain in early embryonic stages, but few factors that drive the migration of yolk sac macrophages into the embryonic brain, or regulate their acquisition of specialized properties are currently known.\n\nHere, we present a CRISPR/Cas9-based in vivo reverse genetic screening pipeline to identify new regulators important for microglia development using zebrafish. Zebrafish larvae are particularly suitable due to their external development, transparency, high fecundity and conserved microglia features. We targeted putative microglia regulators, including signature genes and non-cell autonomous factors, by Cas9/gRNA-complex injections, followed by neutral red-based visualization of microglia. Microglia were quantified automatically in 3-day-old larvae using a software tool we called SpotNGlia. We identified that loss of function of the zebrafish homolog of the colony stimulating factor 1 receptor (CSF1R) ligand IL34, caused strongly reduced microglia numbers in early development. Previous studies on the role of the IL34 on microglia development in vivo were ambiguous. Our data, and a concurrent paper, show that in zebrafish, il34 is required during the earliest seeding of the brain by microglia progenitors. Our data also indicate that Il34 is required for distribution to other organs.\n\nPreviously, we showed that csf1ra and csf1rb double mutant zebrafish have no microglia. As there is a moderate effect of il34 on microglia development, relative to the effect csf1r, additional Csf1r-dependent signalling may be needed for establishment of the microglia network. In all, we identified il34 as a modifier of microglia colonization, by affecting distribution of yolk sac macrophages to target organs, validating our reverse genetic screening pipeline in zebrafish which can be used for the identification of additional regulators of microglia development.

developmental biology↗