A Drosophila haemocyte-specific protein, hemolectin, similar to human von Willebrand factor

Goto, Ayumu; Kumagai, Takeshi; Kumagai, Chino; Hirose, Junko; Narita, Hiroshi; Mori, Hitoshi; Kadowaki, Tatsuhiko; Beck, Konrad; Kitagawa, Yasuo

doi:10.1042/0264-6021:3590099

Cited by 80 publications

(77 citation statements)

References 54 publications

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“…23) with UAS-GFP and used it to label all postmitotic neurons for cell sorting. The HML-GFP line was originally generated by A. Goto 14 (Nagoya University, Japan) and is available through the Bloomington Stock Center. We used markers specific to different R cell subtypes.…”

Section: Methodsmentioning

confidence: 99%

“…12) and Kc 9,13 ) have a profile of Dscam isoforms similar to that of S2 cells (data not shown). We isolated hemocytes by FACS from third instar larvae that express GFP under the control of the promoter region of the hemocyte marker hemolectin (hml) 14 . Consistent with a potential hemocyte origin of these cell lines, the spectrum of Dscam isoforms of primary hemocytes was overall similar to and had the same restriction of exon 9 use as the cell lines.…”

Section: Dscam Expression In Fruit Fly Cell Linesmentioning

confidence: 99%

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Stochastic yet biased expression of multiple Dscam splice variants by individual cells

et al. 2004

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The Drosophila melanogaster gene Dscam is essential for axon guidance and has 38,016 possible alternative splice forms. This diversity can potentially be used to distinguish cells. We analyzed the Dscam mRNA isoforms expressed by different cell types and individual cells. The choice of splice variants expressed is regulated both spatially and temporally. Different subtypes of photoreceptors express broad yet distinctive spectra of Dscam isoforms. Single-cell RT-PCR documented that individual cells express several different Dscam isoforms and allowed an estimation of the diversity that is present. For example, we estimate that each R3/R4 photoreceptor cell expresses 14-50 distinct mRNAs chosen from the spectrum of thousands of splice variants distinctive of its cell type. Thus, the Dscam repertoire of each cell is different from those of its neighbors, providing a potential mechanism for generating unique cell identity in the nervous system and elsewhere. To circumvent these technical difficulties, we used an approach that combined a customized oligonucleotide microarray, isolation of distinct populations of cells and sensitive RT-PCR that allowed examination of Dscam expression by single cells. Here we show that Dscam alternative splicing is differentially regulated both during development and in different tissues. A given cell type expresses a broad, yet distinctive, spectrum of splice variants. Individual cells express a unique repertoire of Dscam splice variants. We estimate that individual R3/R4 photoreceptors express 14-50 distinct mRNA molecules chosen from a subset of thousands of Dscam isoforms characteristic of the population. This would ensure that the Dscam repertoire of each cell is different from those of its neighbors. A R T I C L E S RESULTS A microarray for analyzing Dscam alternative splicingThe genomic structure of Dscam 1 is shown in Figure 1a. There are four alternatively spliced exons. In this study we focused on three alternatively spliced exons, which may create many possible isoforms (Fig. 1a): the cluster in exon 4 has 12 alternative exons, the cluster in exon 6 has 48 alternative exons and the cluster in exon 9 has 33 alternative exons. Each of these highly variable exons can form a portion of an immunoglobulin domain in the extracellular region of the protein. We designed a microarray containing probes for all 93 alternative exons in these three clusters. We amplified cloned DNAs (whose identities were confirmed by sequencing) by PCR to create a reference library that equally represents all the alternative exons present in the microarray. To test the specificity of the different probes in the microarray, we used combinations of RNAs in controlled amounts (Fig. 1b). RNA samples were in vitro transcribed from cloned cDNAs containing different combinations of exons 4, 6 and 9. We carried out a quantitative analysis of four independent experiments in which eight different samples were combined in different amounts spanning two orders of magnitude. Throughout this study, we expressed sample...

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Section: Methodsmentioning

confidence: 99%

Section: Dscam Expression In Fruit Fly Cell Linesmentioning

confidence: 99%

Stochastic yet biased expression of multiple Dscam splice variants by individual cells

et al. 2004

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“…Maturing hemocytes are restricted to the cortical zone of primary lymph gland lobes Mature hemocytes have been shown to express several markers, including collagens (Asha et al, 2003;Fessler et al, 1994;Yasothornsrikul et al, 1997), Hemolectin (Goto et al, 2003;Goto et al, 2001), Lozenge (Lebestky et al, 2000), Peroxidasin and P1 antigen (Asha et al, 2003;Vilmos et al, 2004). We found that the expression of the reporter Collagen-gal4 (Cg-gal4) (Asha et al, 2003), which is expressed by both plasmatocytes and crystal cells, is restricted to the periphery of the primary lymph gland lobe ( Fig.…”

Section: Gross Structure Of the Larval Lymph Glandmentioning

confidence: 99%

“…Additionally, lamellocytes express an uncharacterized surface marker called L1 antigen (Asha et al, 2003). All Drosophila hemocytes specifically express the marker Hemese (He) (Kurucz et al, 2003), while a majority of plasmatocytes and crystal cells express the Collagens Viking and Cg25C (Le Parco et al, 1986;Yasothornsrikul et al, 1997) and the Von Willebrand-like factor Hemolectin (Hml) (Goto et al, 2003;Goto et al, 2001;Sinenko and Mathey-Prevot, 2004).…”

Section: Introductionmentioning

confidence: 99%

TheDrosophilalymph gland as a developmental model of hematopoiesis

et al. 2005

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Drosophila hematopoiesis occurs in a specialized organ called the lymph gland. In this systematic analysis of lymph gland structure and gene expression, we define the developmental steps in the maturation of blood cells (hemocytes) from their precursors. In particular, distinct zones of hemocyte maturation, signaling and proliferation in the lymph gland during hematopoietic progression are described. Different stages of hemocyte development have been classified according to marker expression and placed within developmental niches: a medullary zone for quiescent prohemocytes, a cortical zone for maturing hemocytes and a zone called the posterior signaling center for specialized signaling hemocytes. This establishes a framework for the identification of Drosophila blood cells, at various stages of maturation, and provides a genetic basis for spatial and temporal events that govern hemocyte development. The cellular events identified in this analysis further establish Drosophila as a model system for hematopoiesis.Key words: Drosophila, Hematopoiesis, Blood, Hemocyte, Lymph gland SummaryThe Drosophila lymph gland as a developmental model of hematopoiesis

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“…In Drosophila, undifferentiated prohemocyte progenitors and three differentiated blood cell lineages are being distinguished (Rizki, 1978;Shrestha and Gateff, 1982;Lanot et al, 2001;Hartenstein, 2006). Macrophages, or plasmatocytes, that express Peroxidasin (Pxn) (Nelson et al, 1994;Tepass et al, 1994;Stramer et al, 2005) and Hemolectin (Hml) (Goto et al, 2001;Sinenko and Mathey-Prevot, 2004), correspond to vertebrate myeloid cells and represent 90-95% of hemocytes at most developmental stages (Tepass et al, 1994;Lanot et al, 2001;. Crystal cells and lamellocytes are invertebrate-specific, minor cell types limited to specific developmental stages and immune challenges (Lanot et al, 2001;Crozatier et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

The peripheral nervous system supports blood cell homing and survival in theDrosophilalarva

et al. 2011

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SUMMARYInteractions of hematopoietic cells with their microenvironment control blood cell colonization, homing and hematopoiesis. Here, we introduce larval hematopoiesis as the first Drosophila model for hematopoietic colonization and the role of the peripheral nervous system (PNS) as a microenvironment in hematopoiesis. The Drosophila larval hematopoietic system is founded by differentiated hemocytes of the embryo, which colonize segmentally repeated epidermal-muscular pockets and proliferate in these locations. Importantly, we show that these resident hemocytes tightly colocalize with peripheral neurons and we demonstrate that larval hemocytes depend on the PNS as an attractive and trophic microenvironment. atonal (ato) mutant or genetically ablated larvae, which are deficient for subsets of peripheral neurons, show a progressive apoptotic decline in hemocytes and an incomplete resident hemocyte pattern, whereas supernumerary peripheral neurons induced by ectopic expression of the proneural gene scute (sc) misdirect hemocytes to these ectopic locations. This PNS-hematopoietic connection in Drosophila parallels the emerging role of the PNS in hematopoiesis and immune functions in vertebrates, and provides the basis for the systematic genetic dissection of the PNS-hematopoietic axis in the future.

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A Drosophila haemocyte-specific protein, hemolectin, similar to human von Willebrand factor

Cited by 80 publications

References 54 publications

Stochastic yet biased expression of multiple Dscam splice variants by individual cells

Stochastic yet biased expression of multiple Dscam splice variants by individual cells

TheDrosophilalymph gland as a developmental model of hematopoiesis

The peripheral nervous system supports blood cell homing and survival in theDrosophilalarva

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