Stem Cells and Tissue Repair
We are interested in the cellular mechanisms that underlie polarity and
cell growth during development. Our research is focused on
understanding how maternal components are made and delivered to oocytes
during Drosophila oogenesis. Using mutants with incomplete oocyte
growth, we have discovered key roles for the actin cytoskeleton. For
example, the ring canals connecting growing oocytes with their nurse
cells are stabilized by a special population of bundled actin
The dramatic growth of ring canals during oogenesis requires both actin polymerization and depolymerization, making ring canals a valuable model for in vivo actin dynamics. The polarized movement of maternal mRNA and protein through ring canals from nurse cells to the oocyte is highly regulated. We identified proteins specifically targeted to the oocyte by GFP protein trapping; we are determining the mechanism of targeting using both live imaging and molecular dissection of the proteins to identify localization signals.
Extensive Research Description
Gametes are the ultimate stem cells with the capacity to produce entire
new organisms. We study cellular mechanisms of gamete development using
Drosophila as a model system. We are focused on the development of
female germline cells, from their early differentiation into oocytes or
nurse cells, through the control of oocyte growth during oogenesis. In
addition, we study the role of ovarian muscles in the progression of
developing egg chambers through the ovary.
Early germline development in animals, including flies, relies on a non-canonical form of mitosis. Daughters of germline stem cells undergo a tightly controlled number of mitotic cell divisions with incomplete cytokinesis so that bridges of cytoplasm remain to connect clusters of sister cells. These residual connections are transformed into stable intercellular junctions called ring canals, which are needed for oocyte growth.
In females, this transformation involves recruiting a highly dynamic actin cytoskeleton and many associated actin-binding proteins. Using a variety of genetic and molecular approaches, we have identified many ring canal proteins, and we are actively working toward characterizing their functions. We are also studying the role of ring canals in the polarized transport of maternal mRNAs, proteins and organelles from nurse cells and to the oocyte.
While ring canals are ubiquitous in germline cells, their presence and function in somatic cells are largely unexplored. In order to understand how these fascinating structures contribute to the biology of non-germline cells, we are characterizing somatic ring canals in epithelial cells of the Drosophila ovary and imaginal discs using cell biology and genetics.
Recently we discovered a novel muscle type in the Drosophila ovary that contains striated sarcomeres, but only a single nucleus. This indicates the muscles did not form by typical myoblast fusion. Importantly, the presence of one nucleus means we can use powerful genetic clonal analysis to analyze the effects of mutations affecting muscle proteins, including those associated with human musclular dystrophy. In addition, we can study proliferation of these muscles in adults and the pool of progenitor stem cells that supply new muscle cells in adults.
- Hudson, A.M. and Cooley, L. (2014) Methods for studying oogenesis. In: Drosophila Developmental Biology Methods. Elsevier.
- Yan, D., Neumüller, R., Buckner, M., Ayers, K., Li, H., Hu, Y., Yang-Zhou, D., Pan, L., Wang, X., Kelley, C., Vinayagam, A., Binari, R., Randklev, S., Perkins, L.A., Xie, T., Cooley, L, and Perrimon, N. (2014) A regulatory network of Drosophila germline s
- McLean, P. and Cooley, L. (2013) Protein equilibration through somatic ring canals in Drosophila. Science 340: 1445-1447.
- Neelakanta, G., Hudson, A.M., Sultana, H., Cooley, L. and Fikrig, E. (2012) Expression of Ixodes scapularis antifreeze glycoprotein enhances cold tolerance in Drsosophila melanogaster. (2012) PLoS One 7: e33447.
- Airoldi, S.J., McLean, P., Shimada, Y and Cooley, L. (2011) Intercellular protein movement in syncytial Drosophila follicle cells. J. Cell Sci. 124:4077-4086.
- Shimada, Y., Burn, K.M., Niwa, R. and Cooley, L. (2011) Reversible response of polarized transport to nutrient stress during Drosophila early oogenesis. Devel. Biol., 355:250-262.
- Hudson, A. and Cooley, L. (2010) Drosophila Kelch functions with Cullin-3 to organize the ring canal cytoskeleton. J. Cell Biol. 188: 29-37.
- Hudson, A.H. and Cooley, L. (2008) Phylogenetic, Structural and Functional Relationships between WD- and Kelch-Repeat Proteins. In The Coronin Family of Proteins: Subcellular Biochemistry. Clemen, C., Eichinger, L. and Rybakin, V (eds.) Landes Bioscience
- Hudson, A.H., Petrella, L.N., Tanaka, A.J. and Cooley, L. (2008) Mononuclear muscle cells in Drosophila ovaries revealed by GFP protein traps. Devel. Biol. 314:329-340.
- Petrella, L., Smith-Leiker, T., and Cooley, L. (2007). The Ovhts polyprotein produces fusome and ring canal proteins required for Drosophila oogenesis. Dev. 134:703-712.