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First published online 20 August 2003
doi: 10.1242/dev.00702

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The two origins of hemocytes in Drosophila

Anne Holz*, Barbara Bossinger, Thomas Strasser, Wilfried Janning and Robert Klapper{dagger}

Institut für Allgemeine Zoologie und Genetik der Westfälischen Wilhelms-Universität, Schloßplatz 5, 48149 Münster, Germany



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  		Fig. 1. The embryonic hemocyte (EH) anlage is restricted to 70-80% EL. Clones in third instar larvae after homotopic single-cell transplantation within the head mesoderm. (A) Clone fraction labelling several hemocytes situated in the abdomen of a larva resulting from a homotopic transplantation at 75% EL. (B) Besides their morphology, the labeled cells (blue nuclei) were identified as hemocytes by their characteristic expression of peroxidasin (brown cells). (C) In many cases, labeled hemocytes were detected on the eye-antenna disc of dissected third instar larvae. (D) Transplantation anterior to 85% EL gave rise to endodermal clones. The midgut clone consists of six larval cells (arrows) and many additionally labeled imaginal cells and was obtained after a homotopic single cell transplantation at 89% EL. (E) The EH anlage is embedded into the mesodermal anlage, giving rise to somatic muscles, like the clone in the large head-retractor muscles, deriving from a transplantation at 68% EL.

 


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  		Fig. 2. Heterotopic transplantation reveals the determination of hemocytes at the blastoderm stage. Clones labelling hemocytes in third instar larvae after heterotopic transplantation of single cells from 70-80% EL into the abdominal mesoderm anlage. (A) Fraction of labeled hemocytes intermingled with additional, non-labeled hemocytes derived from a transplantation of a cell originating from 76% EL to 43% EL. (B) Like labeled hemocytes from homotopic transplantations, the hemocytes derived from heterotopic transplantations are also found to be widely dispersed in larvae, as evidenced by this clone fraction in the abdomen.

 


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  		Fig. 3. Embryonic hemocytes (EH) persist through all stages of development. Individual clone after homotopic transplantation of a single GFP-labeled cell at 74% EL at different developmental stages. (A) Several scattered GFP-expressing cells are detectable in a stage 17 embryo. (B) In the third instar larva many dispersed hemocytes either circulate or are attached to the integument. (C) Hemocytes are also detectable in the prepupa. (D) EH persist through metamorphosis, as revealed by the presence of many GFP-expressing cells in the head of the 14-day-old fly.

 


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  		Fig. 4. The lymph glands release hemocytes at the onset of metamorphosis. (A-G) Clones after homotopic transplantations of a single cell (A) or about 10 cells (B-G) between 50 and 55% EL. (A) Lymph gland clone (arrowhead) overlapping with dorsal somatic muscles (arrow) in a third instar larva. (B,C) GFP labelling in a lymph gland lobe of a third instar larva (B, epifluorescence; C, merged with bright-field image). No labeled hemocytes were detected outside of the lymph gland. (D,E) Labelling of a lymph gland lobe (arrow) and longitudinal visceral muscles (arrowheads) in the third instar larva (D, epifluorescence; E, merged with bright-field image). (F) The same specimen as in D at the prepupa stage: several hemocytes are detected either circulating within the hemolymph or sessile on the integument. (G) After metamorphosis, many labeled hemocytes are visible in the adult fly (same specimen as in D and F).

 


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  		Fig. 5. Blastoderm fate map of the embryonic hemocytes (EH) and lymph gland-derived hemocytes (LGH). Within the mesoderm anlage (yellow), which is represented by the ventral region of the blastoderm embryo reaching from 85 to 5% EL, we could identify two anlagen giving rise to hemocytes (red). The EH anlage, which is already determined at the blastoderm stage, is embedded into the head mesoderm and restricted to 70-80% EL. The anlage of the LGH is located at 50-55% EL. Cells giving rise to the lymph glands, and therefore to the LGH, are not determined towards a tissue-specific fate prior to the second postblastodermal mitosis.

 

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© The Company of Biologists Ltd 2003