Overexpression of Fringe Protein in Osteoclast Macrophages
Faculty Mentor
Jason Ashley
Document Type
Poster
Start Date
10-5-2023 11:15 AM
End Date
10-5-2023 1:00 PM
Location
PUB NCR
Department
Biology
Abstract
Osteoclast cells are large multinucleated phagocytic cells that play a crucial role in the reabsorption or breakdown of bone tissue in addition to their role in bone remodeling, growth, and repair. Osteoclastogenesis, the differentiation of osteoclast cells from the precursory line of monocyte/macrophage lineage, is dependent on RANK/RANKL pathway signaling while other pathways, such as Notch signaling, influence other factors of the cell life such as size, activity, and lifespan. Notch pathway signaling is modulated by Fringe glycosyltransferases and has previously been shown to play a role in osteoclast differentiation and function. We will use mouse bone marrow cell lines obtained from the cDNA library cell lines that have been treated to isolate for LFNG, MFNG and RFNG overexpressing cells to analyze the impact of overexpression to the cell. We will use a combination of molecular biology techniques, such as RT-qPCR to verify overexpression, fluorescent viability staining using Propidium Iodide and Hoescht dye, and staining for Tartrate-Resistant Acid Phosphatase (TRAP) activity, to investigate the expression levels of the Fringe proteins and the downstream impact to these cell lines. With the results of this study we seek to provide a foundation for future investigations into the regulation of this critical pathway.
Recommended Citation
Kennerly, Sylvia, "Overexpression of Fringe Protein in Osteoclast Macrophages" (2023). 2023 Symposium. 6.
https://dc.ewu.edu/srcw_2023/res_2023/p2_2023/6
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Overexpression of Fringe Protein in Osteoclast Macrophages
PUB NCR
Osteoclast cells are large multinucleated phagocytic cells that play a crucial role in the reabsorption or breakdown of bone tissue in addition to their role in bone remodeling, growth, and repair. Osteoclastogenesis, the differentiation of osteoclast cells from the precursory line of monocyte/macrophage lineage, is dependent on RANK/RANKL pathway signaling while other pathways, such as Notch signaling, influence other factors of the cell life such as size, activity, and lifespan. Notch pathway signaling is modulated by Fringe glycosyltransferases and has previously been shown to play a role in osteoclast differentiation and function. We will use mouse bone marrow cell lines obtained from the cDNA library cell lines that have been treated to isolate for LFNG, MFNG and RFNG overexpressing cells to analyze the impact of overexpression to the cell. We will use a combination of molecular biology techniques, such as RT-qPCR to verify overexpression, fluorescent viability staining using Propidium Iodide and Hoescht dye, and staining for Tartrate-Resistant Acid Phosphatase (TRAP) activity, to investigate the expression levels of the Fringe proteins and the downstream impact to these cell lines. With the results of this study we seek to provide a foundation for future investigations into the regulation of this critical pathway.
