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 (YSMs) into the embryonic brain, or regulate their acquisition...
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| Format: | Article |
| Language: | English |
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The Company of Biologists
2019-03-01
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| Series: | Disease Models & Mechanisms |
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| Online Access: | http://dmm.biologists.org/content/12/3/dmm037762 |
| _version_ | 1818259188036403200 |
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| author | Laura E. Kuil Nynke Oosterhof Samuël N. Geurts Herma C. van der Linde Erik Meijering Tjakko J. van Ham |
| author_facet | Laura E. Kuil Nynke Oosterhof Samuël N. Geurts Herma C. van der Linde Erik Meijering Tjakko J. van Ham |
| author_sort | Laura E. Kuil |
| collection | DOAJ |
| description | 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 (YSMs) into the embryonic brain, or regulate their acquisition of specialized properties, are currently known. Here, we present a CRISPR/Cas9-based in vivo reverse genetic screening pipeline to identify new microglia regulators using zebrafish. Zebrafish larvae are particularly suitable due to their external development, transparency and conserved microglia features. We targeted putative microglia regulators, 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 zebrafish colony-stimulating factor 1 receptor (Csf1r) ligand, Il34, caused reduced microglia numbers. Previous studies on the role of IL34 in 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. Our data also indicate that Il34 is required for YSM distribution to other organs. Disruption of the other Csf1r ligand, Csf1, did not reduce microglia numbers in mutants, whereas overexpression increased the number of microglia. This shows that Csf1 can influence microglia numbers, but might not be essential for the early seeding of the brain. In all, we identified il34 as a modifier of microglia colonization, by affecting distribution of YSMs to target organs, validating our reverse genetic screening pipeline in zebrafish. This article has an associated First Person interview with the joint first authors of the paper. |
| first_indexed | 2024-12-12T18:11:28Z |
| format | Article |
| id | doaj.art-8e394d2385964afda6c5e0cd578cd7ba |
| institution | Directory Open Access Journal |
| issn | 1754-8403 1754-8411 |
| language | English |
| last_indexed | 2024-12-12T18:11:28Z |
| publishDate | 2019-03-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Disease Models & Mechanisms |
| spelling | doaj.art-8e394d2385964afda6c5e0cd578cd7ba2022-12-22T00:16:23ZengThe Company of BiologistsDisease Models & Mechanisms1754-84031754-84112019-03-0112310.1242/dmm.037762037762Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brainLaura E. Kuil0Nynke Oosterhof1Samuël N. Geurts2Herma C. van der Linde3Erik Meijering4Tjakko J. van Ham5 Department of Clinical Genetics, Erasmus University Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands Department of Clinical Genetics, Erasmus University Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands Biomedical Imaging Group Rotterdam, Departments of Medical Informatics and Radiology, Erasmus University Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands Department of Clinical Genetics, Erasmus University Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands Biomedical Imaging Group Rotterdam, Departments of Medical Informatics and Radiology, Erasmus University Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands Department of Clinical Genetics, Erasmus University Medical Center, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands 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 (YSMs) into the embryonic brain, or regulate their acquisition of specialized properties, are currently known. Here, we present a CRISPR/Cas9-based in vivo reverse genetic screening pipeline to identify new microglia regulators using zebrafish. Zebrafish larvae are particularly suitable due to their external development, transparency and conserved microglia features. We targeted putative microglia regulators, 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 zebrafish colony-stimulating factor 1 receptor (Csf1r) ligand, Il34, caused reduced microglia numbers. Previous studies on the role of IL34 in 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. Our data also indicate that Il34 is required for YSM distribution to other organs. Disruption of the other Csf1r ligand, Csf1, did not reduce microglia numbers in mutants, whereas overexpression increased the number of microglia. This shows that Csf1 can influence microglia numbers, but might not be essential for the early seeding of the brain. In all, we identified il34 as a modifier of microglia colonization, by affecting distribution of YSMs to target organs, validating our reverse genetic screening pipeline in zebrafish. This article has an associated First Person interview with the joint first authors of the paper.http://dmm.biologists.org/content/12/3/dmm037762Reverse genetic screenMicrogliaBrain developmentMacrophagesHematopoiesis |
| spellingShingle | Laura E. Kuil Nynke Oosterhof Samuël N. Geurts Herma C. van der Linde Erik Meijering Tjakko J. van Ham Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brain Disease Models & Mechanisms Reverse genetic screen Microglia Brain development Macrophages Hematopoiesis |
| title | Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brain |
| title_full | Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brain |
| title_fullStr | Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brain |
| title_full_unstemmed | Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brain |
| title_short | Reverse genetic screen reveals that Il34 facilitates yolk sac macrophage distribution and seeding of the brain |
| title_sort | reverse genetic screen reveals that il34 facilitates yolk sac macrophage distribution and seeding of the brain |
| topic | Reverse genetic screen Microglia Brain development Macrophages Hematopoiesis |
| url | http://dmm.biologists.org/content/12/3/dmm037762 |
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