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. 2004 Jun;167(2):707–723. doi: 10.1534/genetics.103.023184

A misexpression screen reveals effects of bag-of-marbles and TGF beta class signaling on the Drosophila male germ-line stem cell lineage.

Cordula Schulz 1, Amy A Kiger 1, Salli I Tazuke 1, Yukiko M Yamashita 1, Luiz C Pantalena-Filho 1, D Leanne Jones 1, Cricket G Wood 1, Margaret T Fuller 1
PMCID: PMC1470893  PMID: 15238523

Abstract

Male gametes are produced throughout reproductive life by a classic stem cell mechanism. However, little is known about the molecular mechanisms for lineage production that maintain male germ-line stem cell (GSC) populations, regulate mitotic amplification divisions, and ensure germ cell differentiation. Here we utilize the Drosophila system to identify genes that cause defects in the male GSC lineage when forcibly expressed. We conducted a gain-of-function screen using a collection of 2050 EP lines and found 55 EP lines that caused defects at early stages of spermatogenesis upon forced expression either in germ cells or in surrounding somatic support cells. Most strikingly, our analysis of forced expression indicated that repression of bag-of-marbles (bam) expression in male GSC is important for male GSC survival, while activity of the TGF beta signal transduction pathway may play a permissive role in maintenance of GSCs in Drosophila testes. In addition, forced activation of the TGF beta signal transduction pathway in germ cells inhibits the transition from the spermatogonial mitotic amplification program to spermatocyte differentiation.

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Selected References

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

  1. Arora K., Dai H., Kazuko S. G., Jamal J., O'Connor M. B., Letsou A., Warrior R. The Drosophila schnurri gene acts in the Dpp/TGF beta signaling pathway and encodes a transcription factor homologous to the human MBP family. Cell. 1995 Jun 2;81(5):781–790. doi: 10.1016/0092-8674(95)90539-1. [DOI] [PubMed] [Google Scholar]
  2. Brand A. H., Perrimon N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development. 1993 Jun;118(2):401–415. doi: 10.1242/dev.118.2.401. [DOI] [PubMed] [Google Scholar]
  3. Chen Dahua, McKearin Dennis M. A discrete transcriptional silencer in the bam gene determines asymmetric division of the Drosophila germline stem cell. Development. 2003 Mar;130(6):1159–1170. doi: 10.1242/dev.00325. [DOI] [PubMed] [Google Scholar]
  4. Chen Dahua, McKearin Dennis. Dpp signaling silences bam transcription directly to establish asymmetric divisions of germline stem cells. Curr Biol. 2003 Oct 14;13(20):1786–1791. doi: 10.1016/j.cub.2003.09.033. [DOI] [PubMed] [Google Scholar]
  5. Fabrizio James J., Boyle Monica, DiNardo Stephen. A somatic role for eyes absent (eya) and sine oculis (so) in Drosophila spermatocyte development. Dev Biol. 2003 Jun 1;258(1):117–128. doi: 10.1016/s0012-1606(03)00127-1. [DOI] [PubMed] [Google Scholar]
  6. FlyBase Consortium The FlyBase database of the Drosophila genome projects and community literature. Nucleic Acids Res. 2003 Jan 1;31(1):172–175. doi: 10.1093/nar/gkg094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gönczy P., DiNardo S. The germ line regulates somatic cyst cell proliferation and fate during Drosophila spermatogenesis. Development. 1996 Aug;122(8):2437–2447. doi: 10.1242/dev.122.8.2437. [DOI] [PubMed] [Google Scholar]
  8. Gönczy P., Matunis E., DiNardo S. bag-of-marbles and benign gonial cell neoplasm act in the germline to restrict proliferation during Drosophila spermatogenesis. Development. 1997 Nov;124(21):4361–4371. doi: 10.1242/dev.124.21.4361. [DOI] [PubMed] [Google Scholar]
  9. Gönczy P., Viswanathan S., DiNardo S. Probing spermatogenesis in Drosophila with P-element enhancer detectors. Development. 1992 Jan;114(1):89–98. doi: 10.1242/dev.114.1.89. [DOI] [PubMed] [Google Scholar]
  10. Hardy R. W., Tokuyasu K. T., Lindsley D. L., Garavito M. The germinal proliferation center in the testis of Drosophila melanogaster. J Ultrastruct Res. 1979 Nov;69(2):180–190. doi: 10.1016/s0022-5320(79)90108-4. [DOI] [PubMed] [Google Scholar]
  11. Hime G. R., Brill J. A., Fuller M. T. Assembly of ring canals in the male germ line from structural components of the contractile ring. J Cell Sci. 1996 Dec;109(Pt 12):2779–2788. doi: 10.1242/jcs.109.12.2779. [DOI] [PubMed] [Google Scholar]
  12. Hinz U., Giebel B., Campos-Ortega J. A. The basic-helix-loop-helix domain of Drosophila lethal of scute protein is sufficient for proneural function and activates neurogenic genes. Cell. 1994 Jan 14;76(1):77–87. doi: 10.1016/0092-8674(94)90174-0. [DOI] [PubMed] [Google Scholar]
  13. Hummel Thomas, Attix Suzanne, Gunning Dorian, Zipursky S. Lawrence. Temporal control of glial cell migration in the Drosophila eye requires gilgamesh, hedgehog, and eye specification genes. Neuron. 2002 Jan 17;33(2):193–203. doi: 10.1016/s0896-6273(01)00581-5. [DOI] [PubMed] [Google Scholar]
  14. Kiger A. A., Jones D. L., Schulz C., Rogers M. B., Fuller M. T. Stem cell self-renewal specified by JAK-STAT activation in response to a support cell cue. Science. 2001 Dec 21;294(5551):2542–2545. doi: 10.1126/science.1066707. [DOI] [PubMed] [Google Scholar]
  15. Kiger A. A., White-Cooper H., Fuller M. T. Somatic support cells restrict germline stem cell self-renewal and promote differentiation. Nature. 2000 Oct 12;407(6805):750–754. doi: 10.1038/35037606. [DOI] [PubMed] [Google Scholar]
  16. Klingler M., Gergen J. P. Regulation of runt transcription by Drosophila segmentation genes. Mech Dev. 1993 Sep;43(1):3–19. doi: 10.1016/0925-4773(93)90019-t. [DOI] [PubMed] [Google Scholar]
  17. Letsou A., Arora K., Wrana J. L., Simin K., Twombly V., Jamal J., Staehling-Hampton K., Hoffmann F. M., Gelbart W. M., Massagué J. Drosophila Dpp signaling is mediated by the punt gene product: a dual ligand-binding type II receptor of the TGF beta receptor family. Cell. 1995 Mar 24;80(6):899–908. doi: 10.1016/0092-8674(95)90293-7. [DOI] [PubMed] [Google Scholar]
  18. Matunis E., Tran J., Gönczy P., Caldwell K., DiNardo S. punt and schnurri regulate a somatically derived signal that restricts proliferation of committed progenitors in the germline. Development. 1997 Nov;124(21):4383–4391. doi: 10.1242/dev.124.21.4383. [DOI] [PubMed] [Google Scholar]
  19. McKearin D. M., Spradling A. C. bag-of-marbles: a Drosophila gene required to initiate both male and female gametogenesis. Genes Dev. 1990 Dec;4(12B):2242–2251. doi: 10.1101/gad.4.12b.2242. [DOI] [PubMed] [Google Scholar]
  20. McKearin D., Ohlstein B. A role for the Drosophila bag-of-marbles protein in the differentiation of cystoblasts from germline stem cells. Development. 1995 Sep;121(9):2937–2947. doi: 10.1242/dev.121.9.2937. [DOI] [PubMed] [Google Scholar]
  21. Ohlstein B., McKearin D. Ectopic expression of the Drosophila Bam protein eliminates oogenic germline stem cells. Development. 1997 Sep;124(18):3651–3662. doi: 10.1242/dev.124.18.3651. [DOI] [PubMed] [Google Scholar]
  22. Rørth P. A modular misexpression screen in Drosophila detecting tissue-specific phenotypes. Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12418–12422. doi: 10.1073/pnas.93.22.12418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rørth P., Szabo K., Bailey A., Laverty T., Rehm J., Rubin G. M., Weigmann K., Milán M., Benes V., Ansorge W. Systematic gain-of-function genetics in Drosophila. Development. 1998 Mar;125(6):1049–1057. doi: 10.1242/dev.125.6.1049. [DOI] [PubMed] [Google Scholar]
  24. Schulz Cordula, Wood Cricket G., Jones D. Leanne, Tazuke Salli I., Fuller Margaret T. Signaling from germ cells mediated by the rhomboid homolog stet organizes encapsulation by somatic support cells. Development. 2002 Oct;129(19):4523–4534. doi: 10.1242/dev.129.19.4523. [DOI] [PubMed] [Google Scholar]
  25. Spradling A., Drummond-Barbosa D., Kai T. Stem cells find their niche. Nature. 2001 Nov 1;414(6859):98–104. doi: 10.1038/35102160. [DOI] [PubMed] [Google Scholar]
  26. Tulina N., Matunis E. Control of stem cell self-renewal in Drosophila spermatogenesis by JAK-STAT signaling. Science. 2001 Dec 21;294(5551):2546–2549. doi: 10.1126/science.1066700. [DOI] [PubMed] [Google Scholar]
  27. Van Doren M., Williamson A. L., Lehmann R. Regulation of zygotic gene expression in Drosophila primordial germ cells. Curr Biol. 1998 Feb 12;8(4):243–246. doi: 10.1016/s0960-9822(98)70091-0. [DOI] [PubMed] [Google Scholar]
  28. Watt F. M., Hogan B. L. Out of Eden: stem cells and their niches. Science. 2000 Feb 25;287(5457):1427–1430. doi: 10.1126/science.287.5457.1427. [DOI] [PubMed] [Google Scholar]
  29. Xu T., Rubin G. M. Analysis of genetic mosaics in developing and adult Drosophila tissues. Development. 1993 Apr;117(4):1223–1237. doi: 10.1242/dev.117.4.1223. [DOI] [PubMed] [Google Scholar]
  30. Yamashita Yukiko M., Jones D. Leanne, Fuller Margaret T. Orientation of asymmetric stem cell division by the APC tumor suppressor and centrosome. Science. 2003 Sep 12;301(5639):1547–1550. doi: 10.1126/science.1087795. [DOI] [PubMed] [Google Scholar]

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