Abstract
A more complete understanding of the biology of adult stem cells could yield important insights toward devising effective cell-based regenerative therapies to treat disease. The germline stem cells (GSCs) in the fruit fly Drosophila melanogaster are an excellent in vivo model for the study of adult stem cell biology. There is increasing evidence from a growing field that microRNAs (miRNAs) play important roles in controlling many aspects of stem-cell biology. Using straightforward genetic manipulations combined with well-established cell biological analysis techniques, we and others have found that the miRNA pathway regulates the cell division rate of Drosophila GSCs as well as the maintenance of the GSCs in their niche. In this chapter, we offer a detailed, self-contained description of a general method to assess the in vivo functions of miRNAs in the GSCs of the Drosophila ovary.
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References
Fuchs, E., Tumbar, T., and Guasch, G. (2004) Socializing with the neighbors: stem cells and their niche. Cell 116, 769–778.
Ting, A. E., Mays, R. W., Frey, M. R., Hof, W. V., Medicetty, S., and Deans, R. (2008) Therapeutic pathways of adult stem cell repair. Crit Rev Oncol Hematol 65, 81–93.
Yang, Y. M., and Chang, J. W. (2008) Current status and issues in cancer stem cell study. Cancer Invest 26, 741–755.
Fuller, M. T., and Spradling, A. C. (2007) Male and female Drosophila germline stem cells: two versions of immortality. Science. 316, 402–404.
Decotto, E., and Spradling, A. C. (2005) The Drosophila ovarian and testis stem cell niches: Similar somatic stem cells and signals. Dev Cell 9, 501–510.
Dansereau, D. A., and Lasko, P. (2008) The development of germline stem cells in Drosophila. Methods Mol Biol 450, 4503–4526.
Förstemann, K., Tomari, Y., Du, T., Vagin, V. V., Denli, A. M., Bratu, D. P., Klattenhoff, C., Theurkauf, W. E., and Zamore, P. D. (2005) Normal microRNA maturation and germ-line stem cell maintenance requires Loquacious, a double-stranded RNA-binding domain protein. PLoS Biol 3, e236.
Hatfield, S. D., Shcherbata, H. R., Fischer, K. A., Nakahara, K., Carthew, R. W., and Ruohola-Baker, H. (2005) Stem cell division is regulated by the microRNA pathway. Nature 435, 974–978.
Jin, Z., and Xie, T. (2007) Dcr-1 maintains Drosophila ovarian stem cells. Curr Biol 17, 539–544.
Park, J. K., Liu, X., Strauss, T. J., McKearin, D. M., and Liu, Q. (2007) The miRNA pathway intrinsically controls self-renewal of Drosophila germline stem cells. Curr Biol 17, 533–538.
Shcherbata, H. R., Ward, E. J., Fischer, K. A., Yu, J., Reynolds, S. H., Chen, C., Xu, P., Hay, B. A., and Ruohola-Baker, H. (2007) Stage-specific differences in the requirements for germline stem cell maintenance in the Drosophila ovary. Cell Stem Cell 1, 698–709.
Yang, L., Chen, D., Duan, R., Xia, L., Wang, J., Qurashi, A., Jin, P., and Chen, D. (2007) Argonaute 1 regulates the fate of germline stem cells in Drosophila. Development 134, 4265–4272.
Yu, J., Reynolds, S. H., Hatfield, S. D., Shcherbata, H. R., Fischer, K. A., Ward, E. J., Long, D., Ding, Y., and Ruohola-Baker, H. (2009) Dicer-1-dependent Dacapo suppression acts downstream of Insulin receptor in regulating cell division of Drosophila germline stem cells. Development 136, 1497–1507.
Neumüller, R. A., Betschinger, J., Fischer, A., Bushati, N., Poernbacher, I., Mechtler, K., Cohen, S. M., and Knoblich, J. A. (2008) Mei-P26 regulates microRNAs and cell growth in the Drosophila ovarian stem cell lineage. Nature 454, 241–245.
Yang, Y., Xu, S., Xia, L., Wang, J., Wen, S., Jin, P., and Chen, D. (2009) The bantam microRNA is associated with drosophila fragile X mental retardation protein and regulates the fate of germline stem cells. PLoS Genet 5, e1000444.
de Nooij, J. C., Letendre, M. A., and Hariharan, I. K. (1996) A cyclin-dependent kinase inhibitor, Dacapo, is necessary for timely exit from the cell cycle during Drosophila embryogenesis. Cell 87, 1237–1247.
Lane, M. E., Sauer, K., Wallace, K., Jan, Y. N., Lehner, C. F., and Vaessin, H. (1996) Dacapo, a cyclin-dependent kinase inhibitor, stops cell proliferation during Drosophila development. Cell 87, 1225–1235.
Golic, K. G. and Lindquist, S. (1989) The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome. Cell 59, 499–509.
Acknowledgment
The authors thank Karin Fischer for her critical reading of the manuscript. This work was supported by the grants from NIH for HRB.
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Chan, K., Ruohola-Baker, H. (2010). Assessing In Vivo MicroRNA Function in the Germline Stem Cells of the Drosophila Ovary. In: Zhang, B., Stellwag, E. (eds) RNAi and microRNA-Mediated Gene Regulation in Stem Cells. Methods in Molecular Biology, vol 650. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-769-3_15
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DOI: https://doi.org/10.1007/978-1-60761-769-3_15
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