New hemocyte‐specific enhancer‐reporter transgenes for the analysis of hematopoiesis in Drosophila

Abstract

Based on environmental challenges or altered genetic composition, Drosophila larvae can produce up to three types of blood cells that express genetic programs essential for their distinct functions. Using transcriptional enhancers for genes expressed exclusively in plasmatocytes, crystal cells, or lamellocytes, several new hemocyte-specific enhancer-reporter transgenes were generated to facilitate the analysis of Drosophila hematopoiesis. This approach took advantage of fluorescent variants of insulated P-element reporter vectors for multilabeling cell analyses; two additional color variants were generated in these studies. These vectors were successfully used to produce transgenic fly lines that label specific hemocyte lineages with separate colors. Combining three transgene reporters allowed for the unambiguous identification of plasmatocytes, crystal cells, and lamellocytes within a complex hemocyte population. While this work focused on the hematopoietic process, these new vectors can be used to mark multiple cell types or trace complex cell lineages during any chosen aspect of Drosophila development.

Neurofibromatosis-like phenotype in Drosophila caused by lack of glucosylceramide extension

“…Akt1 04226 is a lacZ enhancer trap creating an Akt1 hypomorphic allele (42). eater-GFP (*30, II) and eater-Gal4 (*217D201, II) were from T. Tokusumi (University of Notre Dame, Notre Dame, IN) (15,21). Gli-Gal4 (18,22), from M. Stern (Rice University, Houston, TX), drives expression in perineurial glia (43,44).…”

Cell lineage tracing reveals the plasticity of the hemocyte lineages and of the hematopoietic compartments in Drosophila melanogaster

“…To shed light on the validity of our hypothesis that a fraction of the lamellocytes are derived from plasmatocytes, we traced the hemocyte lineages in immune-induced larvae by using two drivers with plasmatocyte-specific expression: Peroxidasin-GAL4 (Pxn-GAL4) (Stramer et al, 2005) and eater-GAL4 (Tokusumi et al, 2009). More than 98% of the plasmatocytes expressed these drivers (not shown).…”

“…Although the hematopoietic compartments and hemocyte subsets have been defined by the expression patterns of immunological markers and in vivo reporter constructs (Kurucz et al, 2003(Kurucz et al, , 2007a(Kurucz et al, , 2007bHonti et al, 2009;Márkus et al, 2009;Tokusumi et al, 2009), the findings of the experiments carried out so far reflect a transient state of the immune system, and our understanding of the lineage-relationships among hemocyte subsets and the composition of the hematopoietic compartments is therefore fragmentary.…”

The matrix protein Tiggrin regulates plasmatocyte maturation inDrosophilalarva

“…To confirm the results obtained with P1 immunostaining, we examined a second marker: Eater-dsRed. This reporter is driven by an enhancer from the Eater locus, which encodes a phagocytosis receptor expressed specifically in plasmatoctyes (Kocks et al, 2005;Tokusumi et al, 2009). Hml>Tig PLs displayed a strong repression of Eater-dsRed expression (Fig.…”