Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal Transduction
Background/Aims: Osteocytes can sense and respond to extracellular stimuli, including biochemical factors throughout the cell body, dendritic processes, and cilia bending. However, further exploration is required of osteocyte function in response to substrate stiffness, an important passive mechanic...
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2018-11-01
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| Series: | Cellular Physiology and Biochemistry |
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| Online Access: | https://www.karger.com/Article/FullText/495482 |
| _version_ | 1819107352302845952 |
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| author | Demao Zhang Chenchen Zhou Qingxuan Wang Linyi Cai Wei Du Xiaobing Li Xuedong Zhou Jing Xie |
| author_facet | Demao Zhang Chenchen Zhou Qingxuan Wang Linyi Cai Wei Du Xiaobing Li Xuedong Zhou Jing Xie |
| author_sort | Demao Zhang |
| collection | DOAJ |
| description | Background/Aims: Osteocytes can sense and respond to extracellular stimuli, including biochemical factors throughout the cell body, dendritic processes, and cilia bending. However, further exploration is required of osteocyte function in response to substrate stiffness, an important passive mechanical cue at the interface between osteocytes and the extracellular matrix, and the deep bio-mechanism in osteocytes involving mechanosensing of cell behavior. Methods: We fabricated silicon-based elastomer polydimethylsiloxane substrates with different stiffnesses but with the same surface topologies. We then seeded osteocytes onto the substrates to examine their responses. Methodologies used included scanning electron microscopy (SEM) for cell morphology, confocal laser scanning microscopy (CLSM) for protein distribution, western blot for protein levels, co-immunoprecipitation for protein interactions, and quantitative real-time polymerase chain reaction for gene expression. Results: SEM images revealed that substrate stiffness induced a change in osteocyte morphology, and CLSM of F-actin staining revealed that substrate stiffness can alter the cytoskeleton. These results were accompanied by changes in focal adhesion capacity in osteocytes, determined via characterization of vinculin expression and distribution. Furthermore, on the exterior of the cell membrane, fibronectin was altered by substrate stiffness. The fibronectin then induced a change in paxillin on the inner membrane of the cell via protein–protein interaction through transmembrane processing. Paxillin led to changes in connexin 43 via protein–protein binding, thereby influencing osteocyte gap junction elongation. Conclusion: This process -from mechanosensing and mechanotransduction to cell function - not only indicates that the effects of mechanical factors on osteocytes can be directly sensed from the cell body, but also indicates the involvement of paxillin transduction. |
| first_indexed | 2024-12-22T02:52:40Z |
| format | Article |
| id | doaj.art-edd720fcfc8548c48ac3c0b4ebedd5bd |
| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-22T02:52:40Z |
| publishDate | 2018-11-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
| record_format | Article |
| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-edd720fcfc8548c48ac3c0b4ebedd5bd2022-12-21T18:41:22ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782018-11-015131013102610.1159/000495482495482Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal TransductionDemao ZhangChenchen ZhouQingxuan WangLinyi CaiWei DuXiaobing LiXuedong ZhouJing XieBackground/Aims: Osteocytes can sense and respond to extracellular stimuli, including biochemical factors throughout the cell body, dendritic processes, and cilia bending. However, further exploration is required of osteocyte function in response to substrate stiffness, an important passive mechanical cue at the interface between osteocytes and the extracellular matrix, and the deep bio-mechanism in osteocytes involving mechanosensing of cell behavior. Methods: We fabricated silicon-based elastomer polydimethylsiloxane substrates with different stiffnesses but with the same surface topologies. We then seeded osteocytes onto the substrates to examine their responses. Methodologies used included scanning electron microscopy (SEM) for cell morphology, confocal laser scanning microscopy (CLSM) for protein distribution, western blot for protein levels, co-immunoprecipitation for protein interactions, and quantitative real-time polymerase chain reaction for gene expression. Results: SEM images revealed that substrate stiffness induced a change in osteocyte morphology, and CLSM of F-actin staining revealed that substrate stiffness can alter the cytoskeleton. These results were accompanied by changes in focal adhesion capacity in osteocytes, determined via characterization of vinculin expression and distribution. Furthermore, on the exterior of the cell membrane, fibronectin was altered by substrate stiffness. The fibronectin then induced a change in paxillin on the inner membrane of the cell via protein–protein interaction through transmembrane processing. Paxillin led to changes in connexin 43 via protein–protein binding, thereby influencing osteocyte gap junction elongation. Conclusion: This process -from mechanosensing and mechanotransduction to cell function - not only indicates that the effects of mechanical factors on osteocytes can be directly sensed from the cell body, but also indicates the involvement of paxillin transduction.https://www.karger.com/Article/FullText/495482SubstrateStiffnessOsteocyteCytoskeletonConnexin 43Paxillin |
| spellingShingle | Demao Zhang Chenchen Zhou Qingxuan Wang Linyi Cai Wei Du Xiaobing Li Xuedong Zhou Jing Xie Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal Transduction Cellular Physiology and Biochemistry Substrate Stiffness Osteocyte Cytoskeleton Connexin 43 Paxillin |
| title | Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal Transduction |
| title_full | Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal Transduction |
| title_fullStr | Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal Transduction |
| title_full_unstemmed | Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal Transduction |
| title_short | Extracellular Matrix Elasticity Regulates Osteocyte Gap Junction Elongation: Involvement of Paxillin in Intracellular Signal Transduction |
| title_sort | extracellular matrix elasticity regulates osteocyte gap junction elongation involvement of paxillin in intracellular signal transduction |
| topic | Substrate Stiffness Osteocyte Cytoskeleton Connexin 43 Paxillin |
| url | https://www.karger.com/Article/FullText/495482 |
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