The focus of our scientific research is the process of myogenesis, with a specific emphasis on the migration, recognition, adhesion and fusion between myoblasts to form multinucleate myotubes. We have chosen embryonic development of the larval body wall muscles in the fruitfly Drosophila melanogaster as a model system, in an effort to utilize the advantages of a combined genetic and molecular approach. Our experimental plan utilizes genes that are associated with serious defects in myoblast fusion as an entry point to identify interacting proteins and signaling pathways that are important to this process. In the embryo, two distinct populations of myoblasts appear to be involved in formation of these fibers. The first, termed muscle founder cells arise in characteristic and reproducible positions in the embryo and contain information that specifies muscle identity, size, position, and attachment. The second and larger group of cells has been termed the fusion-competent myoblasts. Cell-type specific adhesion molecules that are members of the Ig superfamily mediate recognition and fusion between these two distinct populations of myoblasts, and many of the proposed studies focus on the role of these proteins in activating downstream events. SNS, a cell adhesion molecule that is expressed exclusively on the surface of the fusion competent myoblasts, is essential for recognition of Kirre or Rst-expressing founder cells. It appears to activate a signal transduction pathway through interactions that occur with its cytoplasmic domain. Studies within this proposal are aimed at identifying proteins that interact with SNS and are responsible for downstream events. Studies will examine SNS interactions with other transmembrane proteins as well as with cytoplasmic proteins that include SH2- SH3-domain containing adaptor proteins. MBC, the Drosophila ortholog of Dock180, functions as part of an unconventional bipartite guanine nucleotide exchange factor that appears to function downstream of Kirre in founder cells. MBC and its partner Ced-12 are thought to activate the small GTPase Rac1, which has been implicated as a downstream target of multiple pathways in myoblast fusion. We plan to examine this role in more detail by a combination of genetic interaction and live imaging. Lastly, we will use genetic screens that allow us to identify modulators of the above pathways as well as novel genes that function in independent pathways to regulate fusion. Studies have shown that mammalian orthologs of some of these proteins play critical roles in diverse processes in phagocytosis, cell migration and formation of the kidney slit diaphragm. More recent studies have also implicating some of these genes in myoblast fusion in vertebrates.

Public Health Relevance

In a developing embryo, groups of cells of various types must coordinate their movement, interactions and morphology in order to form functional organs. The process of muscle development requires recognition between different myoblasts and fusion of these cells to form muscle fibers. The focus of our research is to examine the mechanisms and essential genes through which cells find each other in the embryo and initiate the process of fusion.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR044274-15
Application #
8300041
Study Section
Skeletal Muscle and Exercise Physiology Study Section (SMEP)
Program Officer
Boyce, Amanda T
Project Start
1996-07-15
Project End
2014-05-31
Budget Start
2012-06-01
Budget End
2014-05-31
Support Year
15
Fiscal Year
2012
Total Cost
$332,450
Indirect Cost
$123,362
Name
Stowers Institute for Medical Research
Department
Type
DUNS #
614653652
City
Kansas City
State
MO
Country
United States
Zip Code
64110
Kaipa, Balasankara Reddy; Shao, Huanjie; Schafer, Gritt et al. (2013) Dock mediates Scar- and WASp-dependent actin polymerization through interaction with cell adhesion molecules in founder cells and fusion-competent myoblasts. J Cell Sci 126:360-72
Haralalka, Shruti; Cartwright, Heather N; Abmayr, Susan M (2012) Recent advances in imaging embryonic myoblast fusion in Drosophila. Methods 56:55-62
Haralalka, Shruti; Shelton, Claude; Cartwright, Heather N et al. (2011) Asymmetric Mbc, active Rac1 and F-actin foci in the fusion-competent myoblasts during myoblast fusion in Drosophila. Development 138:1551-62
Liu, Ze Cindy; Geisbrecht, Erika R (2011) Moleskin is essential for the formation of the myotendinous junction in Drosophila. Dev Biol 359:176-89
Haralalka, Shruti; Abmayr, Susan M (2010) Myoblast fusion in Drosophila. Exp Cell Res :
Shelton, Claude; Kocherlakota, Kiranmai S; Zhuang, Shufei et al. (2009) The immunoglobulin superfamily member Hbs functions redundantly with Sns in interactions between founder and fusion-competent myoblasts. Development 136:1159-68
Geisbrecht, Erika R; Haralalka, Shruti; Swanson, Selene K et al. (2008) Drosophila ELMO/CED-12 interacts with Myoblast city to direct myoblast fusion and ommatidial organization. Dev Biol 314:137-49
Kocherlakota, Kiranmai S; Wu, Jian-Min; McDermott, Jeffrey et al. (2008) Analysis of the cell adhesion molecule sticks-and-stones reveals multiple redundant functional domains, protein-interaction motifs and phosphorylated tyrosines that direct myoblast fusion in Drosophila melanogaster. Genetics 178:1371-83
Balagopalan, Lakshmi; Chen, Mei-Hui; Geisbrecht, Erika R et al. (2006) The CDM superfamily protein MBC directs myoblast fusion through a mechanism that requires phosphatidylinositol 3,4,5-triphosphate binding but is independent of direct interaction with DCrk. Mol Cell Biol 26:9442-55
Galletta, Brian J; Chakravarti, Malabika; Banerjee, Rakhee et al. (2004) SNS: Adhesive properties, localization requirements and ectodomain dependence in S2 cells and embryonic myoblasts. Mech Dev 121:1455-68

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