The Role of Desmoplakin During Epidermal Development Author: Morgan Van Driest Mentor: Dr. Amanda Dickinson.

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Presentation transcript:

The Role of Desmoplakin During Epidermal Development Author: Morgan Van Driest Mentor: Dr. Amanda Dickinson

Experimental Goal To determine how dysfunctional desmoplakin affects the developing epidermis, specifically in the organism Xenopus laevis. Xenopus laevis (African clawed frog)

Project Overview  Background and Significance  Experiment  Expected Outcome  Discussion

Introduction: What is a desmosome? A type of cell junction Involved in maintaining cell adhesion Loss of function results in disease Image by:

Significance: Skin Diseases associated with the desmosome Pemphigus vulgaris Palmoplantar keratoderma Causes: Failure in cell adhesion Molecular defect: dysfunctional desmosomes Image by: Image by: Whittock et al (2002)

Experimental Hypothesis Hypothesis: Desmoplakin is necessary for epidermal development Image by:

Why choose Xenopus? Free living embryos Development can be monitored at all stages Epidermis is similar to the epidermis of mammalian embryos Image by: Xenbase.org

Experimental Design Plakin Rod Mutant construct In vitro transcription mRNA Microinject into embryo Embryo makes mutant protein Formation of epidermis Look at epidermal cell types using EM Plakin Rod Images by: Xenbase.org

Transmission Electron Microscopy Electrons transmitted through specimen Interactions of electrons within the specimen forms image Two part fixation process 1. Glutaraldehyde 2. Osmium tetroxide Plastic embedding 1. Dehydration of sample using ethanol 2. Embedded in epoxy resin

Goblet Cell Desmosome Multiciliated Cell Proper cell differentiation Two cell types: 1. Secretory 2. Multiciliated Characterization of normal cell differentiation in Xenopus laevis Visualization of Normal Cell Differentiation Image by Navaneetha Krishnan Bharathan & Dickinson Lab, unpublished

Experimental Expectations Cell differentiation will not occur Loss of function of desmosomes may cause failure in cell communication Who am I??

Discussion of Possible Results Understanding cell differentiation may lead to understanding diseases associated with the desmosome Useful for developing treatment and prevention

Questions?