The vacuolar H+ ATPase V0 subunit d2 is associated with chondrocyte hypertrophy and supports chondrocyte differentiation.
Abstract: Chondrocyte hypertrophy makes important contributions to bone development and growth. We have investigated a number of novel cartilage genes identified in a recent transcriptomic study to determine whether they are differentially expressed between different zones of equine foetal growth cartilage. Twelve genes (, , , , , , , , , , and ) were found to be more highly expressed in the zone of hypertrophic chondrocytes than in the reserve or proliferative zones, whereas and were expressed at lower levels in the hypertrophic zone than in the reserve zone. , which encodes vacuolar H ATPase (V-ATPase) V subunit d (ATP6V0D2), was selected for further study. Immunohistochemical analysis of ATP6V0D2 in growth cartilage showed stronger staining in hypertrophic than in reserve zone or proliferative chondrocytes. Expression of ATP6V0D2 mRNA and protein was up-regulated in the mouse chondrocytic ATDC5 cell line by conditions inducing expression of hypertrophy-associated genes including and (differentiation medium). In ATDC5 cells cultured in control medium, knockdown of or inhibition of V-ATPase activity using bafilomycin A1 caused a decrease in expression, and in cells cultured in differentiation medium the two treatments caused a decrease in nuclear area. Inhibition of V-ATPase, but not knockdown, prevented the upregulation of , and by differentiation medium, while knockdown, but not inhibition of V-ATPase, caused an increase in the number of ATDC5 cells cultured in differentiation medium. These observations identify ATP6V0D2 as a novel chondrocyte hypertrophy-associated gene. The results are consistent with roles for V-ATPase, both ATP6V0D2-dependent and -independent, in supporting chondrocyte differentiation and hypertrophy.
Publication Date: 2017-08-18 PubMed ID: 29062863PubMed Central: PMC5647522DOI: 10.1016/j.bonr.2017.08.002Google Scholar: Lookup
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- Journal Article
Summary
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This research investigates the expression of certain genes in cartilage growth and confirms the identification of the ATP6V0D2 gene, associated with the vacuolar H+ ATPase V0 subunit d2, as crucial in chondrocyte hypertrophy – a process vital for bone development. Further analysis reveals that this gene contributes to chondrocyte differentiation as well.
Gene Selection and Analysis
- Through a transcriptomic study, the team identified a variety of cartilage genes that were analyzed to determine differential expression in various segments of equine fetal growth cartilage.
- Twelve genes were observed to display heightened expression in the hypertrophic zone, where the growth of cartilage cells into bone cells occurs, compared to the reserve or proliferative zones.
- The research further zeroed in on the ATP6V0D2 gene, which encodes the vacuolar H+ ATPase (V-ATPase) V0 subunit d2.
- An additional analysis showcased stronger staining of ATP6V0D2 in the hypertrophic region, confirming its heightened activity in chondrocyte hypertrophy compared to other zones of cartilage growth.
Exploring ATP6V0D2 Role in Chondrocyte Hypertrophy and Differentiation
- The expression of ATP6V0D2 mRNA and protein was observed to be up-regulated in the mouse chondrocyte ATDC5 cell line under conditions that prompted the expression of hypertrophy-associated genes.
- When ATP6V0D2 or V-ATPase was inhibited, there was a decrease in COL10A1 expression, a marker of chondrocyte hypertrophy, indicating a potential link between ATP6V0D2, V-ATPase, and chondrocyte hypertrophy.
- Notably, inhibiting V-ATPase, but not ATPV0D2 knockdown, prevented upregulation of key genes, hinting at ATP6V0D2-independent roles for V-ATPase in chondrocyte differentiation and hypertrophy.
Conclusion
- This study identifies ATP6V0D2 gene as a new gene associated with chondrocyte hypertrophy.
- The role of V-ATPase, both ATP6V0D2-dependent and -independent, in promoting chondrocyte differentiation and hypertrophy is confirmed, thus offering new insights into the molecular mechanisms behind cartilage growth and bone development.
Cite This Article
APA
Ayodele BA, Mirams M, Pagel CN, Mackie EJ.
(2017).
The vacuolar H+ ATPase V0 subunit d2 is associated with chondrocyte hypertrophy and supports chondrocyte differentiation.
Bone Rep, 7, 98-107.
https://doi.org/10.1016/j.bonr.2017.08.002 Publication
Researcher Affiliations
- Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria 3010, Australia.
- Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria 3010, Australia.
- Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria 3010, Australia.
- Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria 3010, Australia.
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This article has been cited 3 times.- Kazemi M, Williams JL. Properties of Cartilage-Subchondral Bone Junctions: A Narrative Review with Specific Focus on the Growth Plate.. Cartilage 2021 Dec;13(2_suppl):16S-33S.
- Su H, Tang X, Zhang X, Liu L, Jing L, Pan D, Sun W, He H, Yang C, Zhao D, Zhang H, Qi B. Comparative proteomics analysis reveals the difference during antler regeneration stage between red deer and sika deer.. PeerJ 2019;7:e7299.
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