Connexin 43 Is Necessary for Murine Tendon Enthesis Formation and Response to Loading

Shen, Hua; Schwartz, Andrea G.; Civitelli, Roberto; Thomopoulos, Stavros

doi:10.1002/jbmr.4018

Cited by 12 publications

(5 citation statements)

References 34 publications

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“…The BTI is a typical continuous functionally graded structure that attaches tendon to bone and is vital for musculoskeletal function (24). Due to the particularity and importance of the BTI structure, previous studies have reported the formation of BTI (25,26). However, despite these reports, the function of primary cilia, particularly in the formation process of enthesis, was largely unknown.…”

Section: Discussionmentioning

confidence: 99%

Primary cilium of Prrx1+ cells mediate bone-tendon interface development by regulating cell biology

Guan

Xiao

Chen

et al. 2023

Preprint

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Background Bone-tendon interface (BTI) is a typical fibrocartilaginous insertion consisting of continuous yet functionally graded structure. However, the underlying mechanism of enthesis formation is poorly understood. It was reported that primary cilia sense changes in the microenvironment of the BTI undergoing mechanical stimulation to regulate repairing. The objective of this study is to investigate how primary cilia mediate the enthesis formation. Methods Mice at different ages were used for tracing the spatiotemporal distribution of Prrx1+ cells. Cilium were knocked out at 1, 2, 4, and 6 weeks after birth, and the specimens were harvested at 12 weeks to analyze the effect of cilia deletion on the fate of Prrx1+ cells. Histological analysis and microcomputed tomography were utilized to evaluate the formation of BTI. In vitro, Prrx1+ cells were isolated by cell sorting. The expression level of IFT88 protein was knocked down by shRNA, and it was applied to explore the effect of cilia disruption on the proliferation and differentiation of Prrx1+ cells. NVP-LDE225 was used to diminish the expression levels of the Hedgehog (Hh) signaling pathway, which was performed to understand the effect of Hh signaling pathway on the differentiation capacity of Prrx1+ cells. Results Prrx1+ cells participate in the formation of BTI, and in the early postnatal development, Prrx1+ cells are abundant at the BTI. In the late development, Prrx1+ cells gradually decreased in number until disappeared. Locally at the BTI, knockout of cilia may attenuate the chondrogenic differentiation capacity of Prrx1+ cells. Conditional knockout of cilia in the early development is not conducive to BTI formation, and few are in the middle and late stages. In vitro, the knockdown of IFT88 by shRNA could diminish the proliferation and differentiation ability of Prrx1+ cells. At the same time, inhibiting the Hh signaling pathway also diminish the differentiation ability of Prrx1+ cells. The mentioned results suggested that primary cilia may regulate the biological function of Prrx1+ cells via the Hh signaling pathway localized in itself. Conclusions Primary cilia mediate the biological function of Prrx1+ cells to modulate further BTI formation via the Hh signaling pathway.

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Section: Discussionmentioning

confidence: 99%

Primary cilium of Prrx1+ cells mediate bone-tendon interface development by regulating cell biology

Guan

Xiao

Chen

et al. 2023

Preprint

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“…However, in hyaline cartilage and fibrocartilage, specific connexins, including Cx32, Cx43, Cx45, and Cx46, have been reported to mediate the communication between chondrocytes and to be associated with the normal development of joints and some common diseases, such as OA [ 17 , 68 , 102 - 104 ]. In addition, a critical role for Cx43 has been observed in the mineralized fibrocartilaginous transition of bone attachment points in mouse tendons [ 105 ].…”

Section: Cx43 In Physiological Musculoskeletal Systemmentioning

confidence: 99%

Connexin43 in Musculoskeletal System: New Targets for Development and Disease Progression

An¹,

Zheng²,

Cai³

et al. 2022

Aging and disease

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Connexin43, which is the most highly expressed connexin subtype in the musculoskeletal system, exists in a variety of bone cells, synovial tissue, and cartilage tissue. Connexin43 has been suggested to be a key regulator of bone homeostasis. Studies have shown aberrant Connexin43 expression in musculoskeletal disorders, such as osteoporosis, osteoarthritis, and rheumatoid arthritis. During cellular activities, Connexin43 can participate in the formation of functionally specific gap junctions and hemichannels and can exert independent cellular regulatory and signaling functions through special C-termini. The critical role of Connexin43 in physiological development and disease progression has been gradually revealed. In this article, the function of Connexin43 in musculoskeletal tissues is summarized, revealing the potential role of Connexin43 as a key target in the treatment of related bone and muscle disorders and the need for further discovery.

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“…We observed that decorin was invariably less expressed in day 7 compared to day 3 in the presence of FBS, whilst the opposite was the case for ES, especially in the presence of MMC, suggestive of effective native eTC function maintenance. Connexin 43, a gapjunction-associated protein with crucial function in tendon and tendon enthesis formation, function and response to loading [99,100], was observed at passage 3 and at day 3 and day 5 in FBS (higher synthesis was detected under MMC conditions), and in the presence of ES, at passage 3 and at day 7 (MMC independent synthesis). The decreased synthesis of connexin 43 as a function of time in culture (in FBS) and as function of passaging (in both FBS and ES) may be attributed to the absence of mechanical stimulation in this work, which coincides with previous publications that have shown a mechanical loading regime-, time in culture-and age-dependent connexin 43 synthesis and/or expression [101][102][103][104].…”

Section: Sds-page and Immunofluorescence Analysesmentioning

confidence: 99%

Allogeneic Serum and Macromolecular Crowding Maintain Native Equine Tenocyte Function in Culture

Rampin

Skoufos

Raghunath

et al. 2022

Cells

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The absence of a native extracellular matrix and the use of xenogeneic sera are often associated with rapid tenocyte function losses during in vitro culture. Herein, we assessed the influence of different sera (equine serum and foetal bovine serum) on equine tenocyte morphology, viability, metabolic activity, proliferation and protein synthesis as a function of tissue-specific extracellular matrix deposition (induced via macromolecular crowding), aging (passages 3, 6, 9) and time in culture (days 3, 5, 7). In comparison to cells at passage 3, at day 3, in foetal bovine serum and without macromolecular crowding (traditional equine tenocyte culture), the highest number of significantly decreased readouts were observed for cells in foetal bovine serum, at passage 3, at day 5 and day 7 and without macromolecular crowding. Again, in comparison to traditional equine tenocyte culture, the highest number of significantly increased readouts were observed for cells in equine serum, at passage 3 and passage 6, at day 7 and with macromolecular crowding. Our data advocate the use of an allogeneic serum and tissue-specific extracellular matrix for effective expansion of equine tenocytes.

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Connexin 43 Is Necessary for Murine Tendon Enthesis Formation and Response to Loading

Cited by 12 publications

References 34 publications

Primary cilium of Prrx1+ cells mediate bone-tendon interface development by regulating cell biology

Primary cilium of Prrx1+ cells mediate bone-tendon interface development by regulating cell biology

Connexin43 in Musculoskeletal System: New Targets for Development and Disease Progression

Allogeneic Serum and Macromolecular Crowding Maintain Native Equine Tenocyte Function in Culture

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