Macrophage-specific gene functions in Spi1-directed innate immunity
| Autor: | Anna Zakrzewska, Erica L. Benard, Annemarie H. Meijer, Herman P. Spaink, Oliver W. Stockhammer, Chao Cui |
|---|---|
| Přispěvatelé: | Other departments |
| Rok vydání: | 2010 |
| Předmět: |
Salmonella typhimurium
Receptors CXCR3 Transgene Green Fluorescent Proteins Immunology Pair-rule gene Biology Biochemistry Animals Genetically Modified Transcriptome Proto-Oncogene Proteins Animals RNA Messenger Zebrafish Gene In Situ Hybridization Oligonucleotide Array Sequence Analysis Genetics Gene knockdown SPI1 Innate immune system Reverse Transcriptase Polymerase Chain Reaction Macrophages Protein Tyrosine Phosphatase Non-Receptor Type 6 Gene Expression Regulation Developmental Cell Biology Hematology Zebrafish Proteins biology.organism_classification Immunity Innate Recombinant Proteins Hematopoiesis Gene Knockdown Techniques Salmonella Infections Trans-Activators |
| Zdroj: | ResearcherID Blood, 116(3), e1-11. American Society of Hematology |
| ISSN: | 1528-0020 0006-4971 |
| DOI: | 10.1182/blood-2010-01-262873 |
| Popis: | The Spi1/Pu.1 transcription factor plays a crucial role in myeloid cell development in vertebrates. Despite extensive studies of Spi1, the controlled gene group remains largely unknown. To identify genes dependent on Spi1, we used a microarray strategy using a knockdown approach in zebrafish embryos combined with fluorescence-activated cell sorting of myeloid cells from transgenic embryos. This approach of using knockdowns with specific green fluorescent protein-marked cell types was highly successful in identifying macrophage-specific genes in Spi1-directed innate immunity. We found a gene group down-regulated on spi1 knockdown, which is also enriched in fluorescence-activated cell-sorted embryonic myeloid cells of a spi1:GFP transgenic line. This gene group, representing putative myeloid-specific Spi1 target genes, contained all 5 previously identified Spi1-dependent zebrafish genes as well as a large set of novel immune-related genes. Colocalization studies with neutrophil and macrophage markers revealed that genes cxcr3.2, mpeg1, ptpn6, and mfap4 were expressed specifically in early embryonic macrophages. In a functional approach, we demonstrated that gene cxcr3.2, coding for chemokine receptor 3.2, is involved in macrophage migration to the site of bacterial infection. Therefore, based on our combined transcriptome analyses, we discovered novel early macrophage-specific marker genes, including a signal transducer pivotal for macrophage migration in the innate immune response. |
| Databáze: | OpenAIRE |
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