Timing and Location of the Elastic Extracellular Matrix Protein MAGP During Embryonic Development Justin Harper and Barbara Trask Ph.D.

Ex tra cell u lar ma trix (eks tre sel u lar ma triks) n. Intercellular substance synthesized by cells and in which cells are embedded; comprised of a variable combination of molecules including proteins, glycoproteins and proteoglycans. Precise composition is dependent upon the structure and function of the tissue in which it is found. e.g., collagenous matrices, elastic matrices, etc. aka: connective tissue.

The ECM Performs Multiple Functions Structure Molecular Reservoir Adhesion Motility Mecham and Davis, in: Extracellular Matrix Assembly and Structure.

Function of the Elastic Extracellular Matrix Elastic Matrices function in tissues where repeated cycles of extension and recoil are required. i.e. Lungs, Aorta, etc. Elastic Matrices function in tissues where repeated cycles of extension and recoil are required. i.e. Lungs, Aorta, etc.

The Function of Elastin

Components of Elastic Extracellular Matrices Elastin Elastin Microfibrils Microfibrils Fibrillins Fibrillins Fibulins Fibulins Type VI Collagen Type VI Collagen MAGP MAGP *The function of each microfibrillar protein in respect to this matrix is still undetermined

Diseases of the EEM Disfunctional EEMs are linked with conditions such as Emphysema, Marfan Syndrome, and Abdominal Aortic Aneurisms. Disfunctional EEMs are linked with conditions such as Emphysema, Marfan Syndrome, and Abdominal Aortic Aneurisms.

Model Organism Zebrafish (Danio rerio)

Elastic Matrices and Microfibrils in Zebrafish Ventral AortaCiliary Zonules

Discovering MAGP in Zebrafish We used the described sequence of MAGP from Xenopus laevis (African clawed frog) to engineer primers for MAGP in Danio rerio (Zebrafish) We used the described sequence of MAGP from Xenopus laevis (African clawed frog) to engineer primers for MAGP in Danio rerio (Zebrafish) C primers F Primers

Conservation Between Danio rerio and Xenopus laevis Amino Acid Sequences

Optimization of PCR

Embryonic Stages of Development 8 Somite21 Somite26 hpf42 hpf72 hpf

Embryonic MAGP Expression

An Antisense Probe Recognizes a Single mRNA in 24 Hour Danio rerio Embryos ~1.5 kb

Subsequent Steps Expand the Number of Time Points Expand the Number of Time Points Both before 8 somite and after 72hpf Both before 8 somite and after 72hpf In Situ Hybridization In Situ Hybridization Optimization of the probe concentration and development time Optimization of the probe concentration and development time

Acknowledgements Paul Garrett Paul Garrett Elaine Davis Ph.D., McGill University Elaine Davis Ph.D., McGill University Shawn Tolbert and Jeff Pack Shawn Tolbert and Jeff Pack Weber State University Office of Undergraduate Research Weber State University Office of Undergraduate Research WSU Research Scholarship and Professional Growth Office WSU Research Scholarship and Professional Growth Office Department of Zoology, Weber State University Department of Zoology, Weber State University