Discipline: Biological Sciences
Subcategory: Physiology and Health
Miranda Daniel - Bennett College
Co-Author(s): Deborah Garrity, Colorado State University, Fort Collins, CO
Filamins are large cytoplasmic proteins that cross-link cortical actin into dynamic 3-dimensional structures and transmit extracellular signals through integrin receptors into the cytoplasm. Filamins are Y shaped and actin binding proteins. Each chain of filamins makes up an actin binding domain (Akyurek et al, 2007). In humans there are 3 members in the filamin family; filamin A, B and C. My research will focus on filamin C-a. The study of filamin C is important because when mutations occur in Filamins humans develop skeletal and central nervous disorders as well as cardiac malformations. Periventricular nodular heterotopia (PVNH) was the first human phenotype expressed because of mutations in filamins (Akyurek et al, 2007). The Garrity lab is working to discover if filamin C-a plays a role in dilated cardiomyopathy. Collaborators at Colorado Denver School of Medicine have studied a family with a history of dilated cardiomyopathy. It was proven that members of the family with dilated cardiomyopathy had the allele and gene; filamin c-a. Zebrafish filamin c-a and human filamin c are 80% identical on the amino acid level. Because of this strong similarity zebrafish make an excellent animal mode for the study of the disease; Dilated cardiomyopathy. I hypothesize Filamin C-a is required for heart development in zebrafish embryos and the lack of Filamin C-a will be lethal in zebrafish. After injecting morpholino in dosages of 100,250 and 400 uM , I will determine whether smaller dosages will have little or no effect or will higher dosages will have off target effects. Experimenting with different dosages will help us identify the most efficacious dose of morpholino. Some possible phenotypes may include a decrease in contractibility, morphological defects and reduced blood flow. We discovered that 100 uM morpholino is the most efficacious dose because this dose yielded on target phenotypes while the 400 uM morpholino did not. The 400 uM dose of morpholino yielded off target phenotypes such as gastralative defects. Knocking down the gene Filamin C-a with the use of injected splice site morpholino will allow us to screen for a variety of phenotypes. Once the gene Filamin C-a is knocked down using morpholino we will observe the cardiac development of the zebrafish to determine if filamin c-a is required for cardiac development. Understanding the role of filamin c a in cardiac development of zebrafish will help us determine whether the lack of filamin c-a is the cause of cardiac abnormalities in humans. Dilated cardiomyopathy is an important disease to study because it is the most common form of heart disease affecting adults ranging from ages 20- 60. Dilated cardiomyopathy is a disease that effects the left ventricle of the heart. Dilated cardiomyopathy is caused by genetics and not solely bad eating habits.
Funder Acknowledgement(s): National Science Foundation
Faculty Advisor: Christina Moreira,