Title

The Effect Of PAR-1 Activation On Schwann Cell Mophology In A 3-D Collagen Matrix Model

Author(s)

Mary Lyn Mitchell

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Physiology and Health

Presentation Type

Mentored

Mentor

Mentor: Victoria L. Turgeon, Department of Psychology, Furman University

Abstract

Damage to the axons found in the peripheral nervous system is common as it occurs with contusions and cuts often breaking blood vessels that release thrombin onto its surrounding tissues. Thrombin cleaves off Protease activated receptor 1’s (PAR-1) cap revealing the seven amino acid sequence SFLLRNP which binds to another site on PAR-1 causing its activation. PAR-1 is found Schwann cells which are thought to repair damaged axons. Previous research on PAR-1 receptor activation by SFLLRNP alone in a 2D Schwann cell cultures has shown that activating PAR-1 may increase mitosis and alter cell morphology. However, these 2D cultures limit the Schwann cells from interacting with the axons. This project strives to look further at the relationship between activated PAR-1 and Schwann cell morphology in a 3D culture. The 3D model was achieved by placing Schwann cells in collagen matrices. The collagen matrices were treated with 100nM of SFLLRNP and then were collected at three time points (12, 24, and 48 hours) followed by immunocytochemistry staining with antibodies. They were then assessed for cell shape and number of processes using the antibodies anti-vinculin and TRITC – conjugated Phalloidin, which will show the cytoskeleton of the Schwann cells. It was hypothesized that PAR-1 activation would alter the morphology of Schwann cells. The preliminary data showed that PAR-1 did change the shape of Schwann cells from the natural round shape to more amorphous shapes.

Start Date

4-11-2015 10:45 AM

End Date

4-11-2015 11:00 AM

COinS
 
Apr 11th, 10:45 AM Apr 11th, 11:00 AM

The Effect Of PAR-1 Activation On Schwann Cell Mophology In A 3-D Collagen Matrix Model

Damage to the axons found in the peripheral nervous system is common as it occurs with contusions and cuts often breaking blood vessels that release thrombin onto its surrounding tissues. Thrombin cleaves off Protease activated receptor 1’s (PAR-1) cap revealing the seven amino acid sequence SFLLRNP which binds to another site on PAR-1 causing its activation. PAR-1 is found Schwann cells which are thought to repair damaged axons. Previous research on PAR-1 receptor activation by SFLLRNP alone in a 2D Schwann cell cultures has shown that activating PAR-1 may increase mitosis and alter cell morphology. However, these 2D cultures limit the Schwann cells from interacting with the axons. This project strives to look further at the relationship between activated PAR-1 and Schwann cell morphology in a 3D culture. The 3D model was achieved by placing Schwann cells in collagen matrices. The collagen matrices were treated with 100nM of SFLLRNP and then were collected at three time points (12, 24, and 48 hours) followed by immunocytochemistry staining with antibodies. They were then assessed for cell shape and number of processes using the antibodies anti-vinculin and TRITC – conjugated Phalloidin, which will show the cytoskeleton of the Schwann cells. It was hypothesized that PAR-1 activation would alter the morphology of Schwann cells. The preliminary data showed that PAR-1 did change the shape of Schwann cells from the natural round shape to more amorphous shapes.