Protein Kinase C γ Contributes to Central Sensitization in a Rat Model of Chronic Migraine

Wu, Baixue; Wang, Sha; Qin, Guangcheng; Xie, Jingmei; Tan, Ge; Zhou, Jiying; Chen, Lixue

doi:10.1007/s12031-017-0960-7

Cited by 33 publications

(23 citation statements)

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“…The tissues were immediately stored in liquid nitrogen. Total RNA was extracted as previously described [16] using an RNAiso Plus reagent (TaKaRa, Dalian), and RNA was quantified spectrophotometrically with NanoDrop (Thermo, USA). Then, cDNA was synthesized using the PrimeScript™ RT Reagent Kit (Takara).…”

Section: Methodsmentioning

confidence: 99%

Microglia P2X4 receptor contributes to central sensitization following recurrent nitroglycerin stimulation

Long

Pan

et al. 2018

J Neuroinflammation

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BackgroundThe mechanism underlying migraine chronification remains unclear. Central sensitization may account for this progression. The microglia P2X4 receptor (P2X4R) plays a pivotal role in the central sensitization of inflammatory and neuropathic pain, but there is no information about P2X4R in migraine. Therefore, the aim of this study was to identify the precise role of microglia P2X4R in chronic migraine (CM).MethodsWe used an animal model with recurrent intermittent administration of nitroglycerin (NTG), which closely mimics CM. NTG-induced basal and acute mechanical hypersensitivity were evaluated using the von Frey filament test. Then, we detected Iba1 immunoreactivity (Iba1-IR) and P2X4R expression in the trigeminal nucleus caudalis (TNC). To understand the effect of microglia and P2X4R on central sensitization of CM, we examined whether minocycline, an inhibitor of microglia activation, and 5-BDBD, a P2X4R antagonist, altered NTG-induced mechanical hyperalgesia. In addition, we also evaluated the effect of 5-BDBD on c-Fos and calcitonin gene-related peptide (CGRP) expression within the TNC.ResultsChronic intermittent administration of NTG resulted in acute and chronic basal mechanical hyperalgesia, accompanied with microglia activation and upregulation of P2X4R expression. Minocycline significantly decreased basal pain hypersensitivity but did not alter acute NTG-induced hyperalgesia. Minocycline also reduced microglia activation. 5-BDBD completely blocked the basal and acute hyperalgesia induced by NTG. This effect was associated with a significant inhibition of the NTG-induced increase in c-Fos protein and CGRP release in the TNC.ConclusionsOur results indicate that blocking microglia activation may have an effect on the prevention of migraine chronification. Moreover, we speculate that the P2X4R may be implicated in the microglia-neuronal signal in the TNC, which contributes to the central sensitization of CM.Electronic supplementary materialThe online version of this article (10.1186/s12974-018-1285-3) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Microglia P2X4 receptor contributes to central sensitization following recurrent nitroglycerin stimulation

Long

Pan

et al. 2018

J Neuroinflammation

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“…The PWT were measured at least three times in each animal, with an interval of at least 3 min. The average of the three measurements was considered the PWT [13].…”

Section: Behavioural Testsmentioning

confidence: 99%

Microglia P2X4R-BDNF signalling contributes to central sensitization in a recurrent nitroglycerin-induced chronic migraine model

Long

Pan

et al. 2020

J Headache Pain

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Background: According to our previous study, microglia P2X4 receptors (P2X4Rs) play a pivotal role in the central sensitization of chronic migraine (CM). However, the molecular mechanism that underlies the crosstalk between microglia P2X4Rs and neurons of the trigeminal nucleus caudalis (TNC) is not fully understood. Therefore, the aim of this study is to examine the exact P2X4Rs signalling pathway in the development of central sensitization in a CM animal model. Methods: We used an animal model with recurrent intermittent administration of nitroglycerin (NTG), which closely mimics CM. NTG-induced basal mechanical and thermal hypersensitivity were evaluated using a von Frey filament test and an increasing-temperature hot plate apparatus (IITC). We detected P2X4Rs, brain-derived neurotrophic factor (BDNF) and phosphorylated p38 mitogen-activated protein kinase (p-p38-MAPK) expression profiles in the TNC. We investigated the effects of a P2X4R inhibitor (5-BDBD) and an agonist (IVM) on NTG-induced hyperalgesia and neurochemical changes as well as on the expression of p-p38-MAPK and BDNF. We also detected the effects of a tropomyosin-related kinase B (TrkB) inhibitor (ANA-12) on the CM animal model in vivo. Then, we evaluated the effect of 5-BDBD and SB203580 (a p38-MAPK inhibitors) on the release and synthesis of BDNF in BV2 microglia cells treated with 50 μM adenosine triphosphate (ATP). Results: Chronic intermittent administration of NTG resulted in chronic mechanical and thermal hyperalgesia, accompanied by the upregulation of P2X4Rs and BDNF expression. 5-BDBD or ANA-12 prevented hyperalgesia induced by NTG, which was associated with a significant inhibition of the NTG-induced increase in phosphorylated extracellular regulated protein kinases (p-ERK) and calcitonin gene related peptide (CGRP) release in the TNC. Repeated administration of IVM produced sustained hyperalgesia and significantly increased the levels of pERK and CGRP release in the TNC. Activating P2X4Rs with ATP triggered BDNF release and increased BDNF synthesis in BV2 microglia, and these results were then reduced by 5-BDBD or SB203580. Conclusions: Our results indicated that the P2X4R contributes to the central sensitization of CM by releasing BDNF and promoting TNC neuronal hyper-excitability. Blocking microglia P2X4R-BDNF signalling may have an effect on the prevention of migraine chronification.

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“…The molecular mechanisms underlying PKCγ-mediated pain hypersensitivity have been examined in recent studies. PKCγ in the trigeminal nucleus caudalis participates in the pathogenesis of chronic migraine [27]. In addition, spinal protein kinase C is involved in the induction and maintenance of both the persistent spontaneous flinching reflex and contralateral heat hyperalgesia in rats [28].…”

Section: Discussionmentioning

confidence: 99%

Up-regulation of PKCγ subunits of rACC neurons contributes to the development of pain sensitivity in bone cancer rats

She

Zhang

Wang

et al. 2019

Preprint

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Background: To explore the role of PKCγ subunits in rostral anterior cingulate cortex (rACC) neurons in bone cancer pain (BCP) development in rats. Methods: Healthy female Sprague-Dawley rats were randomly divided into five groups: the blank control (naive), sham operation (sham), BCP, BCP plus empty lentiviral vectors (vehicle) and BCP plus PKCγ/shRNA recombinant lentiviral vectors (LV-PKCγ/shRNA) (PKCγ) groups. The BCP, vehicle and PKCγ groups were intra-tibially injected with an MADB-106 rat mammary carcinoma cell suspension (10 μl, 4.6×106 cells/ml), whereas the sham group was intra-tibially injected with saline (10 μl). The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were evaluated on preoperative day 0 (baseline) and days 3, 7, 14 and 21 after intra-tibial implantation. To downregulate PKCγ subunits in rACC neurons, the PKCγ group received a 10 µl bilateral rACC microinjection of shRNA/PKCγ recombinant lentivirus (1.25×109 TU/ml) on day 10 after intra-tibial implantation. The GFP expression of LV-shRNA/PKCγ was examined by microscopy. The vehicle group was injected with an equal dose of empty lentiviral vectors. Western blot, and immunohistochemical approaches were performed to assess the differential expression of PKCγ subunits in rACC neurons among these groups on postoperative days 7 or 21. Results: The baseline MWT and TWL values did not significantly differ among these five groups (P > 0.05). However, compared with the naive and sham groups, the tumour-bearing rats (BCP , vehicle and PKCγ groups) demonstrated marked mechanical allodynia and thermal hyperalgesia starting on postoperative day 7 following intra-tibial implantation of carcinoma cells (P < 0.05). Western blot on postoperative day 7 confirmed a significant increase in PKCγ expression in rACC neurons in the tumour-bearing rats (P < 0.05). However, from postoperative days 14-21, the LV-shRNA/PKCγ microinjection alleviated mechanical allodynia and thermal hyperalgesia in the PKCγ group (P < 0.05). Furthermore, western blot, and histological examination on postoperative day 21 indicated that PKCγ expression in bilateral rACC neurons was significantly lower in the PKCγ group than in the BCP and vehicle groups (P < 0.05). Conclusion: Upregulation of PKCγ subunits in rACC neurons of rats with bone cancer contributes to BCP development.

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Protein Kinase C γ Contributes to Central Sensitization in a Rat Model of Chronic Migraine

Cited by 33 publications

References 50 publications

Microglia P2X4 receptor contributes to central sensitization following recurrent nitroglycerin stimulation

Microglia P2X4 receptor contributes to central sensitization following recurrent nitroglycerin stimulation

Microglia P2X4R-BDNF signalling contributes to central sensitization in a recurrent nitroglycerin-induced chronic migraine model

Up-regulation of PKCγ subunits of rACC neurons contributes to the development of pain sensitivity in bone cancer rats

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