Journal Article

Osteocyte Sptbn1 Deficiency Alters Cell Survival and Mechanotransduction Following Formation of Plasma Membrane Disruptions (PMD) from Mechanical Loading

Tuladhar, Anik, Stokes, Eric G, Piedra, Victor M, Awad, Mohamed E, McGee-Lawrence, Meghan E, Cooley, Marion A, Garland, Theodore, Dorn, Jennifer, Cantu, Nicholas, Cartelle, Anabel Liyen, Hagan, Mackenzie L, Blumenthal, Daniel, Castro, Alberto A, Johnson, Maribeth, Alhamad, Dima W, Roberts, Rachel L, Hamrick, Mark W, Yu, Kanglun, Montesinos-Cartagena, Marlian, Dickerson, Thomas, Bensreti, Husam, Balayan, Vanshika, Bass, Sarah M, McNeil, Paul L

2024

Overview

We and others have shown that application of high-level mechanical loading promotes the formation of transient plasma membrane disruptions (PMD) which initiate mechanotransduction. We hypothesized that increasing osteocyte cell membrane fragility, by disrupting the cytoskeleton-associated protein β2-spectrin (Sptbn1), could alter osteocytic responses and bone adaptation to loading in a PMD-related fashion. In MLO-Y4 cells, treatment with the spectrin-disrupting agent diamide or knockdown of Sptbn1 via siRNA increased the number of PMD formed by fluid shear stress. Primary osteocytes from an osteocyte-targeted DMP1-Cre Sptbn1 conditional knockout (CKO) model mimicked trends seen with diamide and siRNA treatment and suggested the creation of larger PMD, which repaired more slowly, for a given level of stimulus. Post-wounding cell survival was impaired in all three models, and calcium signaling responses from the wounded osteocyte were mildly altered in Sptbn1 CKO cultures. Although Sptbn1 CKO mice did not demonstrate an altered skeletal phenotype as compared to WT littermates under baseline conditions, they showed a blunted increase in cortical thickness when subjected to an osteogenic tibial loading protocol as well as evidence of increased osteocyte death (increased lacunar vacancy) in the loaded limb after 2 weeks of loading. The impaired post-wounding cell viability and impaired bone adaptation seen with Sptbn1 disruption support the existence of an important role for Sptbn1, and PMD formation, in osteocyte mechanotransduction and bone adaptation to mechanical loading.

• Publisher: Springer Nature B.V
• Subject: Adaptation / Biology / Bone growth / Calcium signalling / Cell culture / Cell membranes / Cell survival / Cell viability / Collagen / Cytoskeleton / Fluid flow / Mechanical loading / Mechanical stimuli / Mechanotransduction / Medical research / Osteocytes / Phenotypes / Plasma / Proteins / Shear stress / siRNA / Spectrin / Wounding