Effects of α2,3 Sialylation on CSF1R Signal Potency
Faculty Mentor
Jason Ashley
Document Type
Oral Presentation
Start Date
10-5-2023 11:35 AM
End Date
10-5-2023 11:55 AM
Location
PUB 319
Department
Biology
Abstract
Increased bone resorption caused by excessive osteoclast activity contributes heavily to the occurrence of fragility fractures as well as the bone destruction caused by inflammatory pathologies such as rheumatoid arthritis. Osteoclastogenesis is dependent upon both survival and proliferation signaling provided by colony stimulating factor 1 (CSF1) and differentiation signals provided by receptor activator of nuclear factor kappa-B ligand (RANKL). There is evidence that CSF1 receptor (CSF1R) is decorated with polysaccharides that terminate in the sugar, sialic acid. I hypothesize that the sialylation status of CSF1R contributes to its signaling potency. To investigate these effects, we will culture primary bone marrow macrophages from mice in a medium containing CSF1 and RANKL to induce osteoclastogenesis. Once osteoclastogenesis is initiated we will treat the osteoclast precursors with either a negative control containing only the base medium, CSF1 and RANKL, or base medium and either a heat inactivated or active α2,3 neuraminidase, which can remove terminal sialic acid residues in a stereo-selective manner. Resulting osteoclasts will be imaged via fluorescence microscopy after being stained with nuclear and cell membrane stains and osteoclast size and number will be measured. Following imaging, cells will be fixed and additionally colorimetrically stained for tartrate-resistant acid phosphatase (TRAP) activity and assessed for aberrant morphology. Differences between treatments will be assessed using a Student’s t-test followed by Bonferroni multiple test correction.
Recommended Citation
Beaulaurier, Eric, "Effects of α2,3 Sialylation on CSF1R Signal Potency" (2023). 2023 Symposium. 6.
https://dc.ewu.edu/srcw_2023/res_2023/os2_2023/6
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Effects of α2,3 Sialylation on CSF1R Signal Potency
PUB 319
Increased bone resorption caused by excessive osteoclast activity contributes heavily to the occurrence of fragility fractures as well as the bone destruction caused by inflammatory pathologies such as rheumatoid arthritis. Osteoclastogenesis is dependent upon both survival and proliferation signaling provided by colony stimulating factor 1 (CSF1) and differentiation signals provided by receptor activator of nuclear factor kappa-B ligand (RANKL). There is evidence that CSF1 receptor (CSF1R) is decorated with polysaccharides that terminate in the sugar, sialic acid. I hypothesize that the sialylation status of CSF1R contributes to its signaling potency. To investigate these effects, we will culture primary bone marrow macrophages from mice in a medium containing CSF1 and RANKL to induce osteoclastogenesis. Once osteoclastogenesis is initiated we will treat the osteoclast precursors with either a negative control containing only the base medium, CSF1 and RANKL, or base medium and either a heat inactivated or active α2,3 neuraminidase, which can remove terminal sialic acid residues in a stereo-selective manner. Resulting osteoclasts will be imaged via fluorescence microscopy after being stained with nuclear and cell membrane stains and osteoclast size and number will be measured. Following imaging, cells will be fixed and additionally colorimetrically stained for tartrate-resistant acid phosphatase (TRAP) activity and assessed for aberrant morphology. Differences between treatments will be assessed using a Student’s t-test followed by Bonferroni multiple test correction.
