Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Jul 18;92(15):7075–7079. doi: 10.1073/pnas.92.15.7075

Absence of yolk sac hematopoiesis from mice with a targeted disruption of the scl gene.

L Robb 1, I Lyons 1, R Li 1, L Hartley 1, F Köntgen 1, R P Harvey 1, D Metcalf 1, C G Begley 1
PMCID: PMC41474  PMID: 7624372

Abstract

The scl gene encodes a basic-helix-loop-helix transcription factor which was identified through its involvement in chromosomal translocations in T-cell leukemia. To elucidate its physiological role, scl was targeted in embryonic stem cells. Mice heterozygous for the scl null mutation were intercrossed and their offspring were genotyped. Homozygous mutant (scl-/-) pups were not detected in newborn litters, and analysis at earlier time points demonstrated that scl-/- embryos were dying around embryonic day 9.5. The scl-/- embryos were pale, edematous, and markedly growth retarded after embryonic day 8.75. Histological studies showed complete absence of recognizable hematopoiesis in the yolk sac of these embryos. Early organogenesis appeared to be otherwise normal. Culture of yolk sac cells of wild-type, heterozygous, and homozygous littermates confirmed the absence of hematopoietic cells in scl-/- yolk sacs. Reverse transcription PCR was used to examine the transcripts of several genes implicated in early hematopoiesis. Transcripts of GATA-1 and PU.1 transcription factors were absent from RNA from scl-/- yolk sacs and embryos. These results implicate scl as a crucial regulator of early hematopoiesis.

Full text

PDF
7075

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aplan P. D., Lombardi D. P., Ginsberg A. M., Cossman J., Bertness V. L., Kirsch I. R. Disruption of the human SCL locus by "illegitimate" V-(D)-J recombinase activity. Science. 1990 Dec 7;250(4986):1426–1429. doi: 10.1126/science.2255914. [DOI] [PubMed] [Google Scholar]
  2. Aplan P. D., Lombardi D. P., Reaman G. H., Sather H. N., Hammond G. D., Kirsch I. R. Involvement of the putative hematopoietic transcription factor SCL in T-cell acute lymphoblastic leukemia. Blood. 1992 Mar 1;79(5):1327–1333. [PubMed] [Google Scholar]
  3. Aplan P. D., Nakahara K., Orkin S. H., Kirsch I. R. The SCL gene product: a positive regulator of erythroid differentiation. EMBO J. 1992 Nov;11(11):4073–4081. doi: 10.1002/j.1460-2075.1992.tb05500.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bash R. O., Crist W. M., Shuster J. J., Link M. P., Amylon M., Pullen J., Carroll A. J., Buchanan G. R., Smith R. G., Baer R. Clinical features and outcome of T-cell acute lymphoblastic leukemia in childhood with respect to alterations at the TAL1 locus: a Pediatric Oncology Group study. Blood. 1993 Apr 15;81(8):2110–2117. [PubMed] [Google Scholar]
  5. Begley C. G., Aplan P. D., Denning S. M., Haynes B. F., Waldmann T. A., Kirsch I. R. The gene SCL is expressed during early hematopoiesis and encodes a differentiation-related DNA-binding motif. Proc Natl Acad Sci U S A. 1989 Dec;86(24):10128–10132. doi: 10.1073/pnas.86.24.10128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Begley C. G., Robb L., Rockman S., Visvader J., Bockamp E. O., Chan Y. S., Green A. R. Structure of the gene encoding the murine SCL protein. Gene. 1994 Jan 28;138(1-2):93–99. doi: 10.1016/0378-1119(94)90787-0. [DOI] [PubMed] [Google Scholar]
  7. Bernard O., Lecointe N., Jonveaux P., Souyri M., Mauchauffé M., Berger R., Larsen C. J., Mathieu-Mahul D. Two site-specific deletions and t(1;14) translocation restricted to human T-cell acute leukemias disrupt the 5' part of the tal-1 gene. Oncogene. 1991 Aug;6(8):1477–1488. [PubMed] [Google Scholar]
  8. Boehm T., Foroni L., Kaneko Y., Perutz M. F., Rabbitts T. H. The rhombotin family of cysteine-rich LIM-domain oncogenes: distinct members are involved in T-cell translocations to human chromosomes 11p15 and 11p13. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4367–4371. doi: 10.1073/pnas.88.10.4367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brown L., Cheng J. T., Chen Q., Siciliano M. J., Crist W., Buchanan G., Baer R. Site-specific recombination of the tal-1 gene is a common occurrence in human T cell leukemia. EMBO J. 1990 Oct;9(10):3343–3351. doi: 10.1002/j.1460-2075.1990.tb07535.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Green A. R., DeLuca E., Begley C. G. Antisense SCL suppresses self-renewal and enhances spontaneous erythroid differentiation of the human leukaemic cell line K562. EMBO J. 1991 Dec;10(13):4153–4158. doi: 10.1002/j.1460-2075.1991.tb04993.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Green A. R., Lints T., Visvader J., Harvey R., Begley C. G. SCL is coexpressed with GATA-1 in hemopoietic cells but is also expressed in developing brain. Oncogene. 1992 Apr;7(4):653–660. [PubMed] [Google Scholar]
  12. Guillemot F., Nagy A., Auerbach A., Rossant J., Joyner A. L. Essential role of Mash-2 in extraembryonic development. Nature. 1994 Sep 22;371(6495):333–336. doi: 10.1038/371333a0. [DOI] [PubMed] [Google Scholar]
  13. Haar J. L., Ackerman G. A. Ultrastructural changes in mouse yolk sac associated with the initiation of vitelline circulation. Anat Rec. 1971 Aug;170(4):437–455. doi: 10.1002/ar.1091700406. [DOI] [PubMed] [Google Scholar]
  14. Hwang L. Y., Siegelman M., Davis L., Oppenheimer-Marks N., Baer R. Expression of the TAL1 proto-oncogene in cultured endothelial cells and blood vessels of the spleen. Oncogene. 1993 Nov;8(11):3043–3046. [PubMed] [Google Scholar]
  15. Jan Y. N., Jan L. Y. HLH proteins, fly neurogenesis, and vertebrate myogenesis. Cell. 1993 Dec 3;75(5):827–830. doi: 10.1016/0092-8674(93)90525-u. [DOI] [PubMed] [Google Scholar]
  16. Kallianpur A. R., Jordan J. E., Brandt S. J. The SCL/TAL-1 gene is expressed in progenitors of both the hematopoietic and vascular systems during embryogenesis. Blood. 1994 Mar 1;83(5):1200–1208. [PubMed] [Google Scholar]
  17. Keller G., Kennedy M., Papayannopoulou T., Wiles M. V. Hematopoietic commitment during embryonic stem cell differentiation in culture. Mol Cell Biol. 1993 Jan;13(1):473–486. doi: 10.1128/mcb.13.1.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kouzarides T., Packham G., Cook A., Farrell P. J. The BZLF1 protein of EBV has a coiled coil dimerisation domain without a heptad leucine repeat but with homology to the C/EBP leucine zipper. Oncogene. 1991 Feb;6(2):195–204. [PubMed] [Google Scholar]
  19. Köntgen F., Stewart C. L. Simple screening procedure to detect gene targeting events in embryonic stem cells. Methods Enzymol. 1993;225:878–890. doi: 10.1016/0076-6879(93)25055-7. [DOI] [PubMed] [Google Scholar]
  20. Lecointe N., Bernard O., Naert K., Joulin V., Larsen C. J., Romeo P. H., Mathieu-Mahul D. GATA-and SP1-binding sites are required for the full activity of the tissue-specific promoter of the tal-1 gene. Oncogene. 1994 Sep;9(9):2623–2632. [PubMed] [Google Scholar]
  21. Moore M. A., Metcalf D. Ontogeny of the haemopoietic system: yolk sac origin of in vivo and in vitro colony forming cells in the developing mouse embryo. Br J Haematol. 1970 Mar;18(3):279–296. doi: 10.1111/j.1365-2141.1970.tb01443.x. [DOI] [PubMed] [Google Scholar]
  22. Mortensen R. M., Conner D. A., Chao S., Geisterfer-Lowrance A. A., Seidman J. G. Production of homozygous mutant ES cells with a single targeting construct. Mol Cell Biol. 1992 May;12(5):2391–2395. doi: 10.1128/mcb.12.5.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mouthon M. A., Bernard O., Mitjavila M. T., Romeo P. H., Vainchenker W., Mathieu-Mahul D. Expression of tal-1 and GATA-binding proteins during human hematopoiesis. Blood. 1993 Feb 1;81(3):647–655. [PubMed] [Google Scholar]
  24. Pevny L., Lin C. S., D'Agati V., Simon M. C., Orkin S. H., Costantini F. Development of hematopoietic cells lacking transcription factor GATA-1. Development. 1995 Jan;121(1):163–172. doi: 10.1242/dev.121.1.163. [DOI] [PubMed] [Google Scholar]
  25. Pevny L., Simon M. C., Robertson E., Klein W. H., Tsai S. F., D'Agati V., Orkin S. H., Costantini F. Erythroid differentiation in chimaeric mice blocked by a targeted mutation in the gene for transcription factor GATA-1. Nature. 1991 Jan 17;349(6306):257–260. doi: 10.1038/349257a0. [DOI] [PubMed] [Google Scholar]
  26. Scott E. W., Simon M. C., Anastasi J., Singh H. Requirement of transcription factor PU.1 in the development of multiple hematopoietic lineages. Science. 1994 Sep 9;265(5178):1573–1577. doi: 10.1126/science.8079170. [DOI] [PubMed] [Google Scholar]
  27. Shivdasani R. A., Mayer E. L., Orkin S. H. Absence of blood formation in mice lacking the T-cell leukaemia oncoprotein tal-1/SCL. Nature. 1995 Feb 2;373(6513):432–434. doi: 10.1038/373432a0. [DOI] [PubMed] [Google Scholar]
  28. Sturm K., Tam P. P. Isolation and culture of whole postimplantation embryos and germ layer derivatives. Methods Enzymol. 1993;225:164–190. doi: 10.1016/0076-6879(93)25013-r. [DOI] [PubMed] [Google Scholar]
  29. Szabó P., Mann J. R. Expression and methylation of imprinted genes during in vitro differentiation of mouse parthenogenetic and androgenetic embryonic stem cell lines. Development. 1994 Jun;120(6):1651–1660. doi: 10.1242/dev.120.6.1651. [DOI] [PubMed] [Google Scholar]
  30. Tanigawa T., Elwood N., Metcalf D., Cary D., DeLuca E., Nicola N. A., Begley C. G. The SCL gene product is regulated by and differentially regulates cytokine responses during myeloid leukemic cell differentiation. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7864–7868. doi: 10.1073/pnas.90.16.7864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tsai F. Y., Keller G., Kuo F. C., Weiss M., Chen J., Rosenblatt M., Alt F. W., Orkin S. H. An early haematopoietic defect in mice lacking the transcription factor GATA-2. Nature. 1994 Sep 15;371(6494):221–226. doi: 10.1038/371221a0. [DOI] [PubMed] [Google Scholar]
  32. Tybulewicz V. L., Crawford C. E., Jackson P. K., Bronson R. T., Mulligan R. C. Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene. Cell. 1991 Jun 28;65(7):1153–1163. doi: 10.1016/0092-8674(91)90011-m. [DOI] [PubMed] [Google Scholar]
  33. Valge-Archer V. E., Osada H., Warren A. J., Forster A., Li J., Baer R., Rabbitts T. H. The LIM protein RBTN2 and the basic helix-loop-helix protein TAL1 are present in a complex in erythroid cells. Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8617–8621. doi: 10.1073/pnas.91.18.8617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wadman I., Li J., Bash R. O., Forster A., Osada H., Rabbitts T. H., Baer R. Specific in vivo association between the bHLH and LIM proteins implicated in human T cell leukemia. EMBO J. 1994 Oct 17;13(20):4831–4839. doi: 10.1002/j.1460-2075.1994.tb06809.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Warren A. J., Colledge W. H., Carlton M. B., Evans M. J., Smith A. J., Rabbitts T. H. The oncogenic cysteine-rich LIM domain protein rbtn2 is essential for erythroid development. Cell. 1994 Jul 15;78(1):45–57. doi: 10.1016/0092-8674(94)90571-1. [DOI] [PubMed] [Google Scholar]
  36. Weiss M. J., Keller G., Orkin S. H. Novel insights into erythroid development revealed through in vitro differentiation of GATA-1 embryonic stem cells. Genes Dev. 1994 May 15;8(10):1184–1197. doi: 10.1101/gad.8.10.1184. [DOI] [PubMed] [Google Scholar]
  37. Weiss M. J., Orkin S. H. GATA transcription factors: key regulators of hematopoiesis. Exp Hematol. 1995 Feb;23(2):99–107. [PubMed] [Google Scholar]
  38. Wulf G. M., Adra C. N., Lim B. Inhibition of hematopoietic development from embryonic stem cells by antisense vav RNA. EMBO J. 1993 Dec 15;12(13):5065–5074. doi: 10.1002/j.1460-2075.1993.tb06200.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES