lmmunofluorescent Analysis of Actin Expression in Genetically Modified vs. Wildtype Murine Ocular Muscle
Location
CSU Ballroom
Start Date
21-4-2008 1:00 PM
End Date
21-4-2008 3:00 PM
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Marilyn C. Hart
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
Actin is a cytoplasmic filament that is present in all eukaryotic cells and contributes to cell shape, cell mobility, and to the organization of certain tissues such as striated muscle. Actin is regulated by specialized accessory proteins_including actin capping protein (CP) that serves to stabilize the molecule and regulate its length. CP consists of two distinct protein subunits, an alpha (α) and a beta (β). Two isoforms of the P (βl and β2) subunit have been identified in vertebrates. Genetically modified mice, engineered by Dr. Hart to express lower levels of Pl in their ocular muscles, lost their ability to open their eyes. We hypothesized that that reduced function in the ocular muscles is due to disruption of actin organization in myofibrils. Characterization of this disruption is key to understanding how reduced levels of β1 affect actin organization, myofibrils, and the cell as a whole. Samples of ocular muscle from both wildtype and genetically modified mice were collected and frozen in liquid nitrogen. Tissue sections, seven micron thick, were prepared using a Leica cryomicrotome, probed with a fluorescent actin antibody, and visualized using an Olympus fluorescent microscope. Digitized images were acquired using Simple PCI software.
lmmunofluorescent Analysis of Actin Expression in Genetically Modified vs. Wildtype Murine Ocular Muscle
CSU Ballroom
Actin is a cytoplasmic filament that is present in all eukaryotic cells and contributes to cell shape, cell mobility, and to the organization of certain tissues such as striated muscle. Actin is regulated by specialized accessory proteins_including actin capping protein (CP) that serves to stabilize the molecule and regulate its length. CP consists of two distinct protein subunits, an alpha (α) and a beta (β). Two isoforms of the P (βl and β2) subunit have been identified in vertebrates. Genetically modified mice, engineered by Dr. Hart to express lower levels of Pl in their ocular muscles, lost their ability to open their eyes. We hypothesized that that reduced function in the ocular muscles is due to disruption of actin organization in myofibrils. Characterization of this disruption is key to understanding how reduced levels of β1 affect actin organization, myofibrils, and the cell as a whole. Samples of ocular muscle from both wildtype and genetically modified mice were collected and frozen in liquid nitrogen. Tissue sections, seven micron thick, were prepared using a Leica cryomicrotome, probed with a fluorescent actin antibody, and visualized using an Olympus fluorescent microscope. Digitized images were acquired using Simple PCI software.
Recommended Citation
Peck, Aaron C.. "lmmunofluorescent Analysis of Actin Expression in Genetically Modified vs. Wildtype Murine Ocular Muscle." Undergraduate Research Symposium, Mankato, MN, April 21, 2008.
https://cornerstone.lib.mnsu.edu/urs/2008/poster-session-B/15
