The role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in mice
Abstract Background C–C chemokine receptor 2 (CCR2) signaling plays a key role in pain associated with experimental murine osteoarthritis (OA) after destabilization of the medial meniscus (DMM). Here, we aimed to assess if CCR2 expressed by intra-articular sensory neurons contributes to knee hyperal...
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BMC
2021-04-01
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| Series: | Arthritis Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13075-021-02486-y |
| _version_ | 1818652812044664832 |
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| author | Shingo Ishihara Alia M. Obeidat David L. Wokosin Dongjun Ren Richard J. Miller Anne-Marie Malfait Rachel E. Miller |
| author_facet | Shingo Ishihara Alia M. Obeidat David L. Wokosin Dongjun Ren Richard J. Miller Anne-Marie Malfait Rachel E. Miller |
| author_sort | Shingo Ishihara |
| collection | DOAJ |
| description | Abstract Background C–C chemokine receptor 2 (CCR2) signaling plays a key role in pain associated with experimental murine osteoarthritis (OA) after destabilization of the medial meniscus (DMM). Here, we aimed to assess if CCR2 expressed by intra-articular sensory neurons contributes to knee hyperalgesia in the early stages of the model. Methods DMM surgery was performed in the right knee of 10-week-old male wild-type (WT), Ccr2 null, or Ccr2 RFP C57BL/6 mice. Knee hyperalgesia was measured using a Pressure Application Measurement device. CCR2 receptor antagonist (CCR2RA) was injected systemically (i.p.) or intra-articularly (i.a.) at different times after DMM to test its ability to reverse knee hyperalgesia. In vivo Ca2+ imaging of the dorsal root ganglion (DRG) was performed to assess sensory neuron responses to CCL2 injected into the knee joint cavity. CCL2 protein in the knee was measured by ELISA. Ccr2 RFP mice and immunohistochemical staining for the pan-neuronal marker, protein gene product 9.5 (PGP9.5), or the sensory neuron marker, calcitonin gene-related peptide (CGRP), were used to visualize the location of CCR2 on intra-articular afferents. Results WT, but not Ccr2 null, mice displayed knee hyperalgesia 2–16 weeks after DMM. CCR2RA administered i.p. alleviated established hyperalgesia in WT mice 4 and 8 weeks after surgery. Intra-articular injection of CCL2 excited sensory neurons in the L4-DRG, as determined by in vivo calcium imaging; responses to CCL2 increased in mice 20 weeks after DMM. CCL2, but not vehicle, injected i.a. rapidly caused transient knee hyperalgesia in naïve WT, but not Ccr2 null, mice. Intra-articular CCR2RA injection also alleviated established hyperalgesia in WT mice 4 and 7 weeks after surgery. CCL2 protein was elevated in the knees of both WT and Ccr2 null mice 4 weeks after surgery. Co-expression of CCR2 and PGP9.5 as well as CCR2 and CGRP was observed in the lateral synovium of naïve mice; co-expression was also observed in the medial compartment of knees 8 weeks after DMM. Conclusions The findings suggest that CCL2-CCR2 signaling locally in the joint contributes to knee hyperalgesia in experimental OA, and it is in part mediated through direct stimulation of CCR2 expressed by intra-articular sensory afferents. |
| first_indexed | 2024-12-17T02:27:57Z |
| format | Article |
| id | doaj.art-f167f2cb2b02483c85aff3a5e8fb88a5 |
| institution | Directory Open Access Journal |
| issn | 1478-6362 |
| language | English |
| last_indexed | 2024-12-17T02:27:57Z |
| publishDate | 2021-04-01 |
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| spelling | doaj.art-f167f2cb2b02483c85aff3a5e8fb88a52022-12-21T22:07:03ZengBMCArthritis Research & Therapy1478-63622021-04-0123111210.1186/s13075-021-02486-yThe role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in miceShingo Ishihara0Alia M. Obeidat1David L. Wokosin2Dongjun Ren3Richard J. Miller4Anne-Marie Malfait5Rachel E. Miller6Department of Internal Medicine, Division of Rheumatology, Rush University Medical CenterDepartment of Internal Medicine, Division of Rheumatology, Rush University Medical CenterDepartment of Physiology, Northwestern UniversityDepartment of Pharmacology, Northwestern UniversityDepartment of Pharmacology, Northwestern UniversityDepartment of Internal Medicine, Division of Rheumatology, Rush University Medical CenterDepartment of Internal Medicine, Division of Rheumatology, Rush University Medical CenterAbstract Background C–C chemokine receptor 2 (CCR2) signaling plays a key role in pain associated with experimental murine osteoarthritis (OA) after destabilization of the medial meniscus (DMM). Here, we aimed to assess if CCR2 expressed by intra-articular sensory neurons contributes to knee hyperalgesia in the early stages of the model. Methods DMM surgery was performed in the right knee of 10-week-old male wild-type (WT), Ccr2 null, or Ccr2 RFP C57BL/6 mice. Knee hyperalgesia was measured using a Pressure Application Measurement device. CCR2 receptor antagonist (CCR2RA) was injected systemically (i.p.) or intra-articularly (i.a.) at different times after DMM to test its ability to reverse knee hyperalgesia. In vivo Ca2+ imaging of the dorsal root ganglion (DRG) was performed to assess sensory neuron responses to CCL2 injected into the knee joint cavity. CCL2 protein in the knee was measured by ELISA. Ccr2 RFP mice and immunohistochemical staining for the pan-neuronal marker, protein gene product 9.5 (PGP9.5), or the sensory neuron marker, calcitonin gene-related peptide (CGRP), were used to visualize the location of CCR2 on intra-articular afferents. Results WT, but not Ccr2 null, mice displayed knee hyperalgesia 2–16 weeks after DMM. CCR2RA administered i.p. alleviated established hyperalgesia in WT mice 4 and 8 weeks after surgery. Intra-articular injection of CCL2 excited sensory neurons in the L4-DRG, as determined by in vivo calcium imaging; responses to CCL2 increased in mice 20 weeks after DMM. CCL2, but not vehicle, injected i.a. rapidly caused transient knee hyperalgesia in naïve WT, but not Ccr2 null, mice. Intra-articular CCR2RA injection also alleviated established hyperalgesia in WT mice 4 and 7 weeks after surgery. CCL2 protein was elevated in the knees of both WT and Ccr2 null mice 4 weeks after surgery. Co-expression of CCR2 and PGP9.5 as well as CCR2 and CGRP was observed in the lateral synovium of naïve mice; co-expression was also observed in the medial compartment of knees 8 weeks after DMM. Conclusions The findings suggest that CCL2-CCR2 signaling locally in the joint contributes to knee hyperalgesia in experimental OA, and it is in part mediated through direct stimulation of CCR2 expressed by intra-articular sensory afferents.https://doi.org/10.1186/s13075-021-02486-yOsteoarthritisHyperalgesiaSensitizationPainCCR2CCL2 |
| spellingShingle | Shingo Ishihara Alia M. Obeidat David L. Wokosin Dongjun Ren Richard J. Miller Anne-Marie Malfait Rachel E. Miller The role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in mice Arthritis Research & Therapy Osteoarthritis Hyperalgesia Sensitization Pain CCR2 CCL2 |
| title | The role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in mice |
| title_full | The role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in mice |
| title_fullStr | The role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in mice |
| title_full_unstemmed | The role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in mice |
| title_short | The role of intra-articular neuronal CCR2 receptors in knee joint pain associated with experimental osteoarthritis in mice |
| title_sort | role of intra articular neuronal ccr2 receptors in knee joint pain associated with experimental osteoarthritis in mice |
| topic | Osteoarthritis Hyperalgesia Sensitization Pain CCR2 CCL2 |
| url | https://doi.org/10.1186/s13075-021-02486-y |
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