The overall goal of this proposal is to gain a better understanding of the developmental processes involved in muscle development, while providing a dynamic learning environment for undergraduate students. In this proposal, we plan to identify and characterize a novel collection of zebrafish motility mutants and identify the defective gene in these mutants. A zebrafish motility mutant, dead elvis (del), was detected by the Lessman lab, located at the University of Memphis, in a small pilot experiment using Computer-Aided Screening (CAS) and Computer- Aided Larval Motility Screens (CALMS) (Lessman, 2002;Lessman, 2004). This novel screening series will be carried out on a larger scale to generate more motility mutations in zebrafish at the University of Tennessee at Chattanooga. The del mutation will be investigated by students to further characterize the mutation and discover its molecular cause. Other students will use the CALMS assay to screen for new motility mutations, then proceed to further characterize these new mutant zebrafish. In depth characterization of these mutations can provide critical information into the development and differentiation of muscle tissues and may provide insight into a variety of human neuromuscular conditions. Abnormal development, or injury of any of the components of the neuromuscular system can lead to congenital diseases and disorders such as muscular dystrophy, myasthenia gravis, myopathies, myoglobinuria, Limb-Girdle Syndromes, ataxias, paralysis and spastic paraparesis (Amack and Mahadevan, 2004: Conti-Fine et al., 2006;Mankodi and Thornton, 2002). We believe that the study of zebrafish motility mutants generated by these experiments will produce important insights into the causes and prevention of muscle-related diseases. The generation and characterization of new motility mutants in zebrafish will be carried out in the three major aims of this proposal.
Aim 1 : Identification of zebrafish motility mutants.
Aim 2 : Characterization of zebrafish motility mutants.
Aim 3 : Genetically map and clone the zebrafish motility mutants.
We will develop and study motility mutations in zebrafish. An initial motility mutation, termed dead elvis, is currently being characterized in our lab by undergraduate students. The project includes formation of a series of new mutations in zebrafish, screened by a novel computer based assay to discover zebrafish motility defects. Phenotypic and genetic characterizations within the project will lead to a better understanding the nature of these mutations. Ultimately, these data will increase our knowledge of muscle development and moreover, will advance an overall mandate of the NICHD, Developmental Biology, Genetics &Teratology (DBGT) Branch: abnormal development that relates to the causes and prevention of congenital and genetic defects. On a smaller scale, this project will also advance a program announcement from the National Center for Research Resources (NCRR), Development of Animal Models and Related Biological Materials For Research (R21).
| Lessman, Charles A; Taylor, Michael R; Orisme, Wilda et al. (2010) Use of flatbed transparency scanners in zebrafish research: versatile and economical adjuncts to traditional imaging tools for the Danio rerio laboratory. Methods Cell Biol 100:295-322 |
