Single-cell analysis of dorsal root ganglia reveals metalloproteinase signaling in satellite glial cells and pain

Tonello, Raquel; Prudente, Arthur Silveira; Lee, Sang Hoon; Cohen, Cinder Faith; Xie, Wenrui; Paranjpe, Aditi; Roh, Jueun; Chung, Grace H.; Strong, Judith A.; Zhang, Junming; Berta, Temugin

doi:10.1016/j.bbi.2023.08.005

Cited by 13 publications

(12 citation statements)

References 91 publications

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“…These findings also necessitate a consideration for the role of satellite glial cell (SGC) signaling in DMI therapeutic effects, as these cells express β2-ARs, demonstrate DMI-sensitive activity ( Tang et al, 2010 ), and, upon activation, lead to neuronal hyperexcitability at least in part through pro-inflammatory cascades ( Takeda et al, 2009 ). While our deconvolution suggested no changes in SGC-related DEGs, this could be due to the presence of various SGC subclasses that respond differently to injury states ( Tonello et al, 2023 ). These considerations emphasize the importance of further work determining the effects of DMI on neuronal cells with and without the presence of non-neuronal cells.…”

Section: Discussionmentioning

confidence: 71%

Desipramine induces anti-inflammatory dorsal root ganglion transcriptional signatures in the murine spared nerve injury model

Serafini,

Ramakrishnan,

Shen

et al. 2024

Neurobiology of Pain

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Section: Discussionmentioning

confidence: 71%

Desipramine induces anti-inflammatory dorsal root ganglion transcriptional signatures in the murine spared nerve injury model

Serafini,

Ramakrishnan,

Shen

et al. 2024

Neurobiology of Pain

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“…Rats were placed in plastic chambers and allowed 30 min for acclimation before the examination. As previously described, 29 the mechanical or thermal stimulation was carried out three times to each hind paw at 5 min intervals.…”

Section: Methodsmentioning

confidence: 99%

Grape seed-derived procyanidins decreases neuropathic pain and nerve regeneration by suppression of toll-like receptor 4-myeloid differentiation factor-88 signaling

Hu,

An,

Zhu

et al. 2024

Mol Pain

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Background:Recent studies have shown that peripheral nerve regeneration process is closely related to neuropathic pain. Toll-like receptor 4 (TLR4) signaling was involved in different types of pain and nerve regeneration. TLR4 induced the recruitment of myeloid differentiation factor-88 adaptor protein (MyD88) and NF-κB-depended transcriptional process in sensory neurons and glial cells, which produced multiple cytokines and promoted the induction and persistence of pain. Our study aimed to investigate procyanidins’s effect on pain and nerve regeneration via TLR4-Myd88 signaling. Methods:Spinal nerve ligation (SNL) model was established to measure the analgesic effect of procyanidins. Anatomical measurement of peripheral nerve regeneration was measured by microscopy and growth associated protein 43 (GAP43) staining. Western blotting and/or immunofluorescent staining were utilized to detect TLR4, myeloid differentiation factor-88 adaptor protein (MyD88), ionized calcium-binding adapter molecule 1 (IBA1) and nuclear factor kappa-B-p65 (NF-κB-p65) expression, as well as the activation of astrocyte and microglia. The antagonist of TLR4 (LPS-RS-Ultra, LRU) were intrathecally administrated to assess the behavioral effects of blocking TLR4 signaling on pain and nerve regeneration Result:Procyanidins reduced mechanical allodynia, thermal hyperalgesia and significantly suppressed the number of nerve fibers regenerated and the degree of myelination in SNL model. Compared with sham group, TLR4, MyD88, IBA1 and phosphorylation of NF-κB-p65 were upregulated in SNL rats which were reversed by procyanidins administration. Additionally, procyanidins also suppressed activation of spinal astrocytes and glial cells. Conclusion:Suppression of TLR4-MyD88 signaling contributes to the alleviation of neuropathic pain and reduction of nerve regeneration by procyanidins.

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“…In our research article, we used this protocol to study the role of matrix metalloproteinases in SGCs and pain ( Tonello et al, 2023 ). Here, we present additional evidence that this protocol is both robust and reproducible by assessing these cultures using molecular and functional approaches.…”

Section: Validation Of Protocolmentioning

confidence: 99%

“…Previous protocols for the dissociation and culture of SGCs have been generated from neonatal and immature tissues, contained neuronal and non-neuronal cells, or mostly used rats ( Chen et al, 2008 ; Belzer et al, 2010 ; de Corato et al, 2011 ; Poulsen et al, 2014 ; Wang et al, 2019 ). Recently, there have been publications using cultured SGCs from adult mice ( Su et al, 2022 ; Tonello et al, 2023 ). However, a detailed and well-characterized protocol for the dissociation and culture of mouse SGCs is still lacking.…”

Section: Introductionmentioning

confidence: 99%

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Dissociation and Culture of Adult Mouse Satellite Glial Cells

Tonello,

Davidson,

Berta

2023

BIO-PROTOCOL

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Satellite glial cells (SGCs) are a type of glial cell population that originates from neural crest cells. They ultimately migrate to surround the cell bodies of neurons in the ganglia of the peripheral nervous system. Under physiological conditions, SGCs perform homeostatic functions by modifying the microenvironment around nearby neurons and provide nutrients, structure, and protection. In recent years, they have gained considerable attention due to their involvement in peripheral nerve regeneration and pain. Although methods for culturing neonatal or rat SGCs have long existed, a well-characterized method for dissociating and culturing adult SGCs from mouse tissues has been lacking until recently. This has impeded further studies of their function and the testing of new therapeutics. This protocol provides a detailed description of how to obtain primary cultures of adult SGCs from mouse dorsal root ganglia in approximately two weeks with over 90% cell purity. We also demonstrate cell purity of these cultures using quantitative real-time RT-PCR and their functional integrity using calcium imaging. Key features • Detailed and simplified protocol to dissociate and culture primary satellite glial cells (SGCs) from adult mice. • Cells are dissociated in approximately 2–3 h and cultured for approximately two weeks. • These SGC cultures allow both molecular and functional studies.

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Single-cell analysis of dorsal root ganglia reveals metalloproteinase signaling in satellite glial cells and pain

Cited by 13 publications

References 91 publications

Desipramine induces anti-inflammatory dorsal root ganglion transcriptional signatures in the murine spared nerve injury model

Desipramine induces anti-inflammatory dorsal root ganglion transcriptional signatures in the murine spared nerve injury model

Grape seed-derived procyanidins decreases neuropathic pain and nerve regeneration by suppression of toll-like receptor 4-myeloid differentiation factor-88 signaling

Dissociation and Culture of Adult Mouse Satellite Glial Cells

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