DOI: 10.1126/science.288.5463.146. Specification of <i>Drosophila</i> Hematopoietic Lineage by Conserved Transcription Factors

Specification of Drosophila Hematopoietic Lineage by Conserved Transcription Factors

10.1126/science.288.5463.146

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Two major classes of cells observed within the Drosophila hematopoietic repertoire are plasmatocytes/macrophages and crystal cells. The transcription factor Lz (Lozenge), which resembles human AML1 (acute myeloid leukemia– 1) protein, is necessary for the development of crystal cells during embryonic and larval hematopoiesis. Another transcription factor, Gcm (glial cells missing), has previously been shown to be required for plasmatocyte development. Misexpression of Gcm causes crystal cells to be transformed into plasmatocytes. The Drosophila GATA protein Srp (Serpent) is required for both Lz and Gcm expression and is necessary for the development of both classes of hemocytes, whereas Lz and Gcm are required in a lineage-specific manner. Given the similarities of Srp and Lz to mammalian GATA and AML1 proteins, observations in Drosophila are likely to have broad implications for understanding mammalian hematopoiesis and leukemias.

Authors 4

Tim Lebestky (first)
Ting Chang (additional)
Volker Hartenstein (additional)
Utpal Banerjee (additional)

References 37 Referenced 426

Orkin S. H., Int. J. Dev. Biol. 42, 927 (1998). / Int. J. Dev. Biol. by Orkin S. H. (1998)
10.1016/S0959-437X(98)80009-9
10.1073/pnas.93.22.12355
10.1182/blood.V89.10.3636
10.1038/384474a0
10.1242/dev.122.12.4023
10.1038/372143a0
10.1073/pnas.93.8.3444
10.1016/S0092-8674(00)80986-1
Speck N. A., et al., Cancer Res. 59, 1789s (1999); / Cancer Res. by Speck N. A. (1999)
Bae S., Ito Y., Histol. Histopathol. 14, 1213 (1999). / Histol. Histopathol. by Bae S. (1999)
10.1101/gad.10.10.1194
10.1242/dev.125.18.3681
10.1006/dbio.1997.8702
10.1016/0092-8674(95)90281-3
10.1016/0092-8674(95)90280-5
10.1242/dev.120.7.1829
10.1242/dev.125.4.713
10.1016/S0092-8674(00)81396-3
Dearolf C. R., Biochim. Biophys. Acta 1377, M13 (1998). / Biochim. Biophys. Acta by Dearolf C. R. (1998)
T. M. Rizki in Genetics and Biology of Drosophila M. Ashburner and T. R. F. Wright Eds. (Academic Press London 1978) vol. 2b pp. 398–451.
10.1007/BF00848799
10.1111/j.1440-169X.1982.00065.x
10.1007/BF00360522
T. Rizki and R. Rizki Genetics 97 s90 (1981).
T. Lebestky T. Chang V. Hartenstein U. Banerjee data not shown.
For further details see Science Online (www.sciencemag.org/feature/data/1047010.shl).
Embryos were collected for 2 hours at either 25°C (for up-shift experiments) or 29°C (for down-shift experiments) and shifted either up or down at stages 8 10 11 13 14 and 15. Completion of embryogenesis takes about 22 hours at 25°C and 16 hours at 29°C. The ratio of 16/22 was used to compensate for this timing differential in the down-shift experiments. When shifted down at stages 8 and 10 black cells were still present within the embryos analyzed at stage 17. When shifted down at stages 11 13 14 and 15 black cells were absent from the embryos at stage 17. When shifted up to 29°C at stages 8 10 11 13 and 14 black cells were absent from the embryos at stage 17 but when shifted up at stage 15 black cells were present in stage 17 embryos. These results suggest that Lz function is required continuously between stages 10 and 14 for crystal cell development. Furthermore when lz ts1 ; Bc/+ embryos develop at 25°C until stage 17 but are then shifted to 29°C during larval stages no crystal cells are observed in the third-instar larvae indicating that embryonic crystal cells do not persist into late larval stages. Additionally when lz ts1 ; Bc/+ embryos are raised at 29°C until third-instar larvae and are shifted to 25°C crystal cells appear within 4.5 hours after the restoration of Lz function.
Antisense lz mRNA probe was made from the 3.5-kb lz cDNA (9). Rabbit antibody to Lz (9) was used at 1:50 dilution. Affinity-purified mouse antibody to Lz was used at 1:100 dilution. In situ hybridization and immunohistochemistry protocols were as described (12).
Lymph glands were dissected by cutting at 7/8 length along the larval body. Using fine tweezers the inner tissue was inverted and lymph glands were separated from surrounding tissue. Lymph glands were fixed and stained using protocols derived from eye disc staining protocols as described (9).
Smears of circulating hemocytes from third-instar hemolymph were created by cutting larvae with fine tweezers and depositing the hemolymph on a glass slide. For immunolocalization smears were fixed in 0.5% glutaraldehyde for 30 s rinsed with PBT (PBS + 0.05% Triton) five times incubated in PBT-N (PBS + 0.05% Triton + 10% normal goat serum) for 1 min incubated in primary antibody overnight at 4°C rinsed in PBT five times washed in PBT-N for 1 min incubated in secondary antibody for 2 hours at room temperature rinsed five times in PBT and mounted in Vectashield (Vector Laboratories) fluorescent mounting medium.
The gcm-lacZ referenced in the text and legends is the gcm rA87 enhancer trap allele. Similarly lz-lacZ refers to the lz-gal4; UAS-lacZ genotype.
10.1073/pnas.95.21.12364
10.1016/S0014-5793(98)01650-0
10.1073/pnas.93.25.14912
10.1126/science.284.5418.1313
We thank R. Reuter for useful discussions the Srp antibody and the srp neo45 allele; L. Fessler and J. Fessler for helpful suggestions and discussions; N. Franc for providing Croquemort antibody; A. Giangrande for useful discussions UAS-gcm and gcm rA87 and Gcm antibody; C. Goodman and B. Jones for Gcm antibody; J. Pollock for the lz-gal4 stock; K. Matthews at the Bloomington Stock Center for many fly stocks; members of the Banerjee lab for critical review and comments on the manuscript; and E. Chan for his involvement in early stages of the project. Supported by U.S. Public Health Service National Research Service Award GM07185 (T.L.); NIH grant RO1EY08152 the McKnight Foundation Investigator Award and the Margaret E. Early Medical Research Trust Award (U.B.); and NIH grant NS29367 (V.H.).

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 5:35 a.m.)
Deposited	1 year, 7 months ago (Jan. 13, 2024, 5:11 a.m.)
Indexed	1 day, 17 hours ago (Aug. 30, 2025, 12:56 p.m.)
Issued	25 years, 4 months ago (April 7, 2000)
Published	25 years, 4 months ago (April 7, 2000)
Published Print	25 years, 4 months ago (April 7, 2000)

Funders 0

None

BibTeX

@article{Lebestky_2000, title={Specification of Drosophila Hematopoietic Lineage by Conserved Transcription Factors}, volume={288}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.288.5463.146}, DOI={10.1126/science.288.5463.146}, number={5463}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Lebestky, Tim and Chang, Ting and Hartenstein, Volker and Banerjee, Utpal}, year={2000}, month=apr, pages={146–149} }

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  "abstract": "<jats:p>\n            Two major classes of cells observed within the\n            <jats:italic>Drosophila</jats:italic>\n            hematopoietic repertoire are plasmatocytes/macrophages and crystal cells. The transcription factor Lz (Lozenge), which resembles human AML1 (acute myeloid leukemia\u2013 1) protein, is necessary for the development of crystal cells during embryonic and larval hematopoiesis. Another transcription factor, Gcm (glial cells missing), has previously been shown to be required for plasmatocyte development. Misexpression of Gcm causes crystal cells to be transformed into plasmatocytes. The\n            <jats:italic>Drosophila</jats:italic>\n            GATA protein Srp (Serpent) is required for both Lz and Gcm expression and is necessary for the development of both classes of hemocytes, whereas Lz and Gcm are required in a lineage-specific manner. Given the similarities of Srp and Lz to mammalian GATA and AML1 proteins, observations in\n            <jats:italic>Drosophila</jats:italic>\n            are likely to have broad implications for understanding mammalian hematopoiesis and leukemias.\n          </jats:p>",
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