Timing and Location of the Timing and Location of the Elastic Extracellular Matrix Elastic Extracellular Matrix

Protein MAGP During Protein MAGP During Embryonic DevelopmentEmbryonic Development

Justin Harper and Barbara Trask Justin Harper and Barbara Trask Ph.D.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 The ECM Performs Multiple FunctionsFunctions

Structure

Molecular Reservoir

Adhesion

Motility

Mecham and Davis, 1994. in: Extracellular Matrix Assembly and Structure.

Function of the Elastic Function of the Elastic Extracellular MatrixExtracellular Matrix

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

The Function of ElastinThe Function of Elastin

Components of Elastic Components of Elastic Extracellular MatricesExtracellular Matrices

ElastinElastin MicrofibrilsMicrofibrils

FibrillinsFibrillins FibulinsFibulins Type VI CollagenType VI Collagen MAGPMAGP

*The function of each microfibrillar protein in *The function of each microfibrillar protein in respect to this matrix is still undeterminedrespect to this matrix is still undetermined

Diseases of the EEMDiseases of the EEM

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

Model OrganismModel OrganismZebrafish (Zebrafish (Danio rerio)Danio rerio)

Elastic Matrices and Microfibrils Elastic Matrices and Microfibrils in Zebrafishin Zebrafish

Ventral Aorta

Ciliary Zonules

Discovering MAGP in Discovering MAGP in ZebrafishZebrafish

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

C primers

F Primers

Conservation Between Conservation Between Danio Danio rerio rerio andand Xenopus laevis Xenopus laevis Amino Amino

Acid SequencesAcid Sequences

Optimization of PCROptimization of PCR

Embryonic Stages of Embryonic Stages of DevelopmentDevelopment

8 Somite 21 Somite 26 hpf 42 hpf 72 hpf

Embryonic MAGP ExpressionEmbryonic MAGP Expression

An Antisense Probe Recognizes An Antisense Probe Recognizes a Single mRNA in 24 Hour a Single mRNA in 24 Hour

Danio rerioDanio rerio Embryos Embryos

~1.5 kb

Subsequent StepsSubsequent Steps Expand the Number of Time PointsExpand the Number of Time Points

Both before 8 somite and after 72hpfBoth before 8 somite and after 72hpf

In Situ In Situ HybridizationHybridization Optimization of the probe concentration Optimization of the probe concentration

and development timeand development time

AcknowledgementsAcknowledgements

Paul GarrettPaul Garrett Elaine Davis Ph.D., McGill UniversityElaine Davis Ph.D., McGill University Shawn Tolbert and Jeff PackShawn Tolbert and Jeff Pack Weber State University Office of Weber State University Office of

Undergraduate ResearchUndergraduate Research WSU Research Scholarship and Professional WSU Research Scholarship and Professional

Growth OfficeGrowth Office Department of Zoology, Weber State Department of Zoology, Weber State

UniversityUniversity