Electrophysiological characterization of the tetrodotoxin-resistant Na+ channel, Nav1.9, in mouse dorsal root ganglion neurons

Small dorsal root ganglion neurons express preferentially the Na⁺ channel isoform Na_V1.9 that mediates a tetrodotoxin-resistant (TTX-R) Na⁺ current. We investigated properties of the Na⁺ current mediated by Na_V1.9 (I_NaN) using the whole-cell, patch-clamp recording technique. To isolate I_NaN from heterogeneous TTX-R Na⁺ currents that also contain another type of TTX-R Na⁺ current mediated by Na_V1.8, we used Na_V1.8-null mutant mice. When F^– was used as an internal anion in the patch pipette solution, both the activation and inactivation kinetics for I_NaN shifted in the hyperpolarizing direction with time. Such a time-dependent shift of the kinetics was not observed when Cl^– was used as an internal anion. Functional expression of I_NaN declined with time after cell dissociation and recovered during culture, implying that Na_V1.9 may be regulated dynamically by trophic factors or depend on subtle environmental factors for its survival. During whole-cell recordings, the peak amplitude of I_NaN increased dramatically after a variable delay, as if inactive or silent channels had been kindled. Such an unusual increase of the amplitude could be prevented by adding ATP to the pipette solution or by recording with the nystatin-perforated patch-clamp technique, suggesting that the rupture of patch membrane affected the behaviour of Na_V1.9. These peculiar properties of I_NaN may provide an insight into the plasticity of Na⁺ channels that are related to pathological functions of Na⁺ channels accompanying abnormal pain states.     

小鼠背根神经节中痒觉神经元的分布特性

目的观察小鼠背根神经节(DRG)中痒觉特异性Mrgpr A3+神经元的分布特征。方法采用遗传学方法将Mrgpr A3+神经元特异性标记强化绿色荧光蛋白(EGFP)和td Tomato;选取3只纯合Mrgpra3EGFP-Cre;ROSA26td Tomato成年转基因小鼠,分离皮肤和背根神经节组织;采用激光扫描共焦成像技术观察Mrgpr A3+神经元的外周神经纤维在小鼠躯体皮肤的投射分布特征;采用双光子成像技术观察Mrgpr A3+神经元在整体背根神经节中的三维空间分布情况。结果脸颊、背部和脚掌皮肤的Mrgpr A3+神经纤维分布密集,粗且长,分布广泛;颈部和腹部皮肤的Mrgpr A3+神经纤维分布稀疏,短且小,呈散点状分布;Mrgpr A3+神经纤维在皮肤的有毛和无毛区域都有投射分布,且不同部位分布特点不同;几乎所有的Mrgpr A3+神经元都为小直径感觉神经元,且在颈、胸、腰、尾段背根神经节中均有分布;颈段、胸段、腰段和尾段的Z轴成像深度分别为350μm、250μm、400μm和200μm;躯体不同部位背根神经节中的Mrgpr A3+神经元的三维空间分布在不同节段存在明显差异。结论小鼠躯体不同部位皮肤的Mrgpr A3+神经纤维的分布特征和整体背根神经节中Mrgpr A3+神经元的三维空间分布特征都存在较大差异,痒觉神经元和末梢分布的差异可能是不同区域存在痒觉生理反应差异的结构基础。     

Differential effects of riluzole on subpopulations of adult rat dorsal root ganglion neurons in vitro

Riluzole is clinically approved for the treatment of motoneuron disease. We have previously shown that this drug is neuroprotective for both sensory neurons and motoneurons and promotes neurite outgrowth [Bergerot A, Shortland PJ, Anand P, Hunt SP, Carlstedt T (2004) Exp Neurol 187(2):359–366; Shortland PJ, Leinster VH, White W, Robson LG (2006) Eur J Neurosci 24:3343–3353]. This study explored the effects of exogenous administration of 0.1 μM riluzole on the neurite growth of specific subpopulations of adult rat dorsal root ganglion (DRG) neurons in vitro. Neuronal branching and neurite length were measured in calcitonin gene related peptide (CGRP), Griffonia simplicifolia Isolectin B4 (IB4), N52 and parvalbumin positive neuronal subpopulations. Riluzole was found to enhance neurite branching in both CGRP and IB4 positive neurons compared to vehicle treated cultures. However, neurite length was only significantly increased in CGRP positive neurons in riluzole treated cultures. The results suggest that riluzole affects specific subpopulations of sensory neurons in vitro and that its effects may be mediated through activation of neurotrophic factor receptors, since neurite outgrowth could be blocked by the administration of K252a (at 10 nM). Riluzole may offer a new pharmacological approach to promote sensory regeneration following small fibre neuropathies.     

Inhibitory action of protein kinase Cbeta inhibitor on tetrodotoxin-resistant Na+ current in small dorsal root ganglion neurons in diabetic rats

Experimental evidence has been presented to suggest that protein kinase Cbeta isoform-selective inhibitor LY333531 is effective at alleviating diabetic hyperalgesia. In the present study, we isolated small (< or =25 microm in soma diameter) dorsal root ganglion (DRG) neurons from control and streptozocin (STZ)-induced diabetic rats, and examined the acute action of LY333531 (1-1000 nM) on the tetrodotoxin-resistant Na(+) current (TTX-R I(Na)), which plays an essential role in transmitting nociceptive impulses, using the whole-cell patch-clamp method. TTX-R I(Na) in diabetic DRG neurons was enhanced in amplitude (71.5+/-3.6pA/pF, n=10 versus 41.2+/-3.3pA/pF, n=8) and was activated at more negative potentials (V(1/2), -15.1+/-1.3 mV versus -9.6+/-1.4 mV), compared with that in control neurons. Bath application of LY333531 acutely inhibited TTX-R I(Na) in both control and diabetic DRG neurons, and the degree of inhibition by the drug at concentrations of 1, 10 and 100 nM was significantly greater in diabetic DRG neurons than in control DRG neurons. Thus, TTX-R I(Na), which is upregulated in the diabetic state, is likely to be more potently inhibited by submicromolar concentrations of LY333531. These results suggest that an acute inhibition of TTX-R I(Na) by LY333531 attenuates the exaggerated excitability of DRG neurons in the diabetic state, which appears to be related at least partly to anti-hyperalgesic actions of the drug in diabetic neuropathy.     

体外原代培养小鼠胚胎背根神经节细胞的神经化学特征 

目的 探讨胚胎背根神经节（DRG）感觉神经元作为体外研究神经多肽的细胞模型之有效性。方法 采用免疫荧光及相差显微技术对30只胚胎小鼠背跟神经节细胞选择性神经多肽的分布、细胞大小及其多肽表达与细胞大小之间的关系进行观察比较。结果 培养时间达3周龄的DRG细胞主要以中小直径（30~20μm）的细胞为主,与成年在体脊髓DRG细胞的形态多形性特征类似;选择性神经多肽（钙调素基因相关多肽、P物质、甘丙肽和 nociceptin）的表达也随着体外培养时间的延长明显增强,且从早期仅在胞体部位表达到3周时细胞周围     

TNF-α在CCD模型大鼠背根神经节中的表达及机制

目的:探讨TNF-α和NF-κB在背根神经节慢性压迫(CCD)模型大鼠背根神经节(DRG)中的表达变化及其对疼痛学行为的影响。方法:建立CCD大鼠模型,采用von Frey纤维丝监测机械痛阈的改变;通过Western blotting检测TNF-α和NF-κB在DRG中的表达变化趋势,分析其与疼痛行为之间的相关性;并采用免疫荧光双染技术研究TNF-α在DRG中的表达位置。结果:CCD组的50%机械缩足阈值在术后1 d即开始明显下降(P0.01),7~14 d达到高峰,其后逐渐上升,直至术后35 d仍明显低于术前及sham组(P0.01)。而DRG上的TNF-α及NF-κB于造模后各时点均显著增多(P0.01),且TNF-α的表达趋势与50%机械缩足阈值显著相关(P0.05)。结论:DRG慢性压迫可促进其上的TNF-α和NF-κB的合成和分泌,进而诱发机械痛觉过敏。因此,TNF-α/NF-κB信号通路可能是CCD模型疼痛形成的重要通路之一。     

Spike potentials and membrane properties of dorsal root ganglion cells in pigeons

Active and passive membrane properties of dorsal root ganglion (DRG)-cells from the intact superfused ganglion of pigeons have been compared with the conduction velocity of their centrifugal axons.About two thirds of the neurones were associated with myelinated axons and classified as A-cells; the remainder were associated with unmyelinated axons and classified as C-cells. Slowly conducting group III A-cells (5–25 m·s^–1) constituted half of the A-cell population. With exception of spike duration, spike parameters and membrane properties did not differ among the A-cells. Spike duration increased with decreasing conduction velocity demonstrating a small plateau (hump) during the fall time in group III neurones. This hump was more distinct in C-cells, resulting in a 2–5 times longer duration of action potentials. Amplitude and duration of afterhyperpolarization (AHP) of C-cells was 2–3 times that of A-cells.Administration of 10 mM CoCl₂ decreased the rate of rise and the overshoot but increased the rate of fall of the action potential in C-cells and group III A-cells, largely abolishing the hump. It is suggested that the hump of the spike potential is largely produced by a Cacurrent and that the resultant increase of intracellular Ca might produce the larger AHP in C-cells, secondary to an increase in K-conductance.     

Type III intermediate filament peripherin inhibits neuritogenesis in type II spiral ganglion neurons in vitro

Peripherin, a type III intermediate filament protein, forms part of the cytoskeleton in a subset of neurons, most of which have peripheral fibre projections. Studies suggest a role for peripherin in axon outgrowth and regeneration, but evidence for this in sensory and brain tissues is limited. The exclusive expression of peripherin in a sub-population of primary auditory neurons, the type II spiral ganglion neurons (SGN) prompted our investigation of the effect of peripherin gene deletion (pphKO) on these neurons. We used confocal immunofluorescence to examine the establishment of the innervation of the cochlear outer hair cells by the type II SGN neurites in vivo and in vitro, in wildtype (WT) and pphKO mice, in the first postnatal week. The distribution of the type II SGN nerve fibres was normal in pphKO cochleae. However, using P1 spiral ganglion explants under culture conditions where the majority of neurites were derived from type II SGN, pphKO resulted in increased numbers of neurites/explant compared to WT controls. Type II SGN neurites from pphKO explants extended ∼double the distance of WT neurites, and had reduced complexity based on greater distance between turning points. Addition of brain-derived neurotrophic factor (BDNF) to the culture media increased neurite number in WT and KO explants ∼30-fold, but did not affect neurite length or distance between turning. These results indicate that peripherin may interact with other cytoskeletal elements to regulate outgrowth of the peripheral neurites of type II SGN, distinguishing these neurons from the type I SGN innervating the inner hair cells.     

二脱氧肌苷引发培养的背根神经节神经元的形态学改变

为探讨二脱氧肌苷(ddI)对培养的背根神经节(DRG)神经元的影响,我们对分散培养的胎鼠DRG神经元培养3d后,再分别以不同浓度的ddI(1μg/ml,5μg/ml,10μg/mll和20μg/ml)孵育3d。终止培养后,行微管相关蛋白2(MAP2)标记,用共聚焦激光扫描显微镜观察神经元胞体和突起的改变。结果表明,DRG神经元用ddI孵育3d,神经元突起的数目减少和长度变短,呈剂量依赖性,而神经元的直径则没有变化。本研究的结果表明,ddI可影响培养的DRG神经元突起的再生和生长。     

猕猴胸腰段脊神经节nNOS免疫阳性神经元的分布

目的　观察正常猕猴胸腰段脊神经节神经元型一氧化氮合酶 (nNOS)免疫阳性神经元的分布。方法　用ABC免疫细胞化学方法显示nNOS免疫阳性神经元 ,并用体视学方法进行定量分析。结果　胸段和腰段脊神经节nNOS免疫阳性神经元的分布相似 ,均可见较丰富的nNOS免疫阳性神经元分布 ,神经元的大小不等 ,多呈圆形或椭圆形。胞浆着色较深 ,胞核位于细胞中央 ,不着色 ,细胞被神经纤维束分隔成群。nNOS免疫阳性神经元以中型神经元为主 ,其次为小型神经元 ,其胞浆呈强阳性染色 ,细胞直径 <50 μm ,大型神经元较少。胸、腰段脊神经节nNOS免疫阳性神经元的密度无明显差异 (P >0 0 5)。结论　在灵长类动物中 ,NO可能在感觉的传导和调节中发挥重要作用 ,但由于nNOS主要在中、小型神经元中表达 ,提示NO可能主要参与痛觉的调制     

Peripheral nerve injury decreases the expression of metabolic glutamate receptor 7 in dorsal root ganglion neurons

Group II and III metabolic glutamate receptors (mGluRs) are responsible for the glutamate-mediated postsynaptic excitation of neurons. Previous pharmacological evidences show that activation of mGluR7 could inhibit nociceptive reception. However, the distribution and expression patterns of mGluR7 after peripheral injury remain unclear. Herein we found that mGluR7 was expressed in the rat peptidergic dorsal root ganglion (DRG) neurons and large neurons, but rarely in isolectin B4 positive neurons. Sciatic nerve ligation experiment showed that mGluR7 was anterogradely transported from cell body to the peripheral site. Furthermore, after peripheral nerve injury, mGluR7 expression was down-regulated in both peptidergic and large DRG neurons. Our work suggests that mGluR7 might be involved in the regulation of pathological pain after peripheral nerve injury.     

Distribution patterns of different P2x receptor phenotypes in acutely dissociated dorsal root ganglion neurons of adult rats

P2x receptors may be used to detect ATP release from tissues during physiological and pathological conditions. We used whole-cell patch clamp recordings to study the expression of P2x receptor phenotypes, their distribution patterns, and their sensitivity to alphabetamATP and suramin in dorsal root ganglion (DRG) neurons acutely dissociated from adult rats. Based on the onset and decay rates of 10 microM ATP-evoked currents, we showed three types of P2x currents: fast, slow, and mixed. Each of these P2x receptor phenotypes had a distinct distribution pattern among DRG neurons. The fast P2x currents were predominantly expressed in small-diameter, isolectin-B4 (IB4)-positive, and capsaicin-sensitive DRG neurons. The slow P2x currents were expressed in both small and medium DRG neurons, and about half of them were IB4 positive. The mixed P2x currents were also expressed in both small and medium-sized DRG neurons, and most of these neurons were IB4-positive neurons. The slow and mixed P2x current groups had both capsaicin-sensitive and -insensitive DRG neurons. All phenotypes revealed with 10 microM ATP could be inhibited by 30 microM suramin. All DRG neurons with fast or mixed P2x currents were also sensitive to 10 microM alphabetamATP, and alphabetamATP evoked currents similar to those induced by ATP. The group expressing slow P2x currents could be further divided into alphabetamATP-sensitive and -insensitive groups. Thus, the relationships among P2x receptor phenotypes, cell sizes, IB4 positivity, and capsaicin sensitivity are more complicated than previously thought, and different P2x receptors may be involved in both nociceptive and non-nociceptive functions.     

The effect of mechanical strain or hypoxia on cell death in subpopulations of rat dorsal root ganglion neurons in vitro

Spinal nerves and their associated dorsal root ganglion (DRG) cells can be subject to mechanical deformation and hypoxia associated with pathology such as disc herniation, spinal stenosis and spine trauma. There is very limited information on the response of adult DRG neurons to such stressors. In this study we used an in vitro approach to examine the response of adult DRG cells to (a) mechanical, hypoxic, and combined injuries; and (b) to compare the effects on injury on nociceptive and non-nociceptive neurons, as well as on non-neuronal cells. Mechanical injury (20% tensile strain) led to significant neuronal cell death (assessed by ethidium homodimer-1 labelling), which was proportional to strain duration (5 min, 1 h, 6 h or 18 h). Hypoxia (2% O₂ for 24 h) also promoted death of DRG neurons, and was further enhanced when mechanical strain and hypoxia were combined. Both mechanical strain and hypoxia significantly decreased the maximum neurite length. Conversely, death of non-neuronal cells was only increased by hypoxia and not by mechanical strain. Total cell death in response to mechanical injury or hypoxia was similar in both non-nociceptive (neurofilament, NF-200 immunoreactive) and nociceptive (calcitonin gene-related peptide, CGRP immunoreactive) neurons, but apoptosis (assessed by activated caspase-3 immunostaining) was significantly higher in CGRP than NF-200 neurons. Surprisingly, cell death of non-peptidergic nociceptors (identified by Griffonia simplicifolia IB4 lectin binding) was already high in control cultures, and was not increased further by either mechanical stretch or hypoxia. These results provide detailed information on the response of adult DRG subpopulations to hypoxia and mechanical strain, and describe in vitro models that could be useful for screening potential neuroprotective agents.     

Comparison of two types of Na+ currents with low-voltage-activated T-type Ca2+ current in newborn rat dorsal root ganglia

Na⁺ currents and the low-voltage-activated T-type Ca²⁺ current (T-I _Ca) were recorded from neurons of rat dorsal root ganglia under similar ionic environments using the whole-cell patch-clamp technique. Two types of Na⁺ currents were identified on the basis of their sensitivity to tetrodotoxin (TTX) and channel kinetics. One type was blocked by 1 nM TTX and had a faster activation and inactivation time courses (F-I _Na), while the other type was insensitive to 100 M TTX and had a much slower channel kinetics (S-I _Na). Activation thresholds were –60, –40 and –70 mV for F-I _Na, S-I _Na and T-I _Ca, respectively. Peak amplitudes were obtained in respective current/voltage curves at –30 mV (F-I _Na), 0 mV (S-I _Na) and –50 mV (T-I _Ca). The time to peak and the decay time constant measured at potential levels giving peak amplitudes were 0.5 and 1.5ms for F-I _Na, 1.4 and 2.9 ms for S-I _Na and 8.1 and 17 ms for T-I _Ca, respectively. Cd²⁺ in a concentration of 50 M totally blocked T-I _Ca, whereas it had no effect on either type of Na⁺ current. T-I _Na was found in 18 out of 25 cells which possessed F-I _Ns, whereas it was found in only 2 cells among 15 which lacked F-I _Na. These three types of inward currents having different kinetic and pharmacological properties may mediate diverse functional roles in processing sensory signals.     

大鼠背根神经节内kv2.1 mRNA和kv4.2 mRNA的表达

目的　了解kv2 1和kv4 2型钾离子通道在背根神经节的表达。方法　应用原位杂交方法观察背根神经节内细胞膜钾离子kv2 1和kv4 2型通道的mRNA阳性神经结构的表达。结果　腰 3～ 5背根神经节内均有kv2 1和kv4 2mRNA阳性神经元的表达 ,kv4 2mRNA阳性神经元的数量和总面积明显多于kv2 1阳性神经元。结论　kv2 1和kv4 2型钾离子通道可能参与了背根神经节对外周传入信息的传递和调制。     

Alteration of dorsal root ganglion P2X3 receptor expression and function following spinal nerve ligation in the rat

One subtype of ATP-gated ion channel, the P2X₃ receptor, is expressed primarily on peripheral sensory neurons. While it is known that P2X₃ receptors can participate in certain forms of nociceptive signaling, their involvement in neuropathic pain transmission is not known. We have examined the expression and function of P2X₃ receptors in a rat spinal nerve ligation model of neuropathic pain. Fourteen days following L5/L6 spinal nerve ligation, the corresponding dorsal root ganglia (DRG) were removed from animals exhibiting mechanical allodynia, and these were studied using immunohistochemical and electrophysiological techniques. Using a polyclonal antibody to label the P2X₃ receptor, a significant reduction in neuronal P2X₃ immunoreactivity was observed in the ipsilateral (injured) L5 and L6 DRG following nerve ligation. In vitro electrophysiological analysis of acutely isolated DRG neurons revealed a similar decrease in functional P2X₃-containing receptors. In small diameter (22–25 μm) neurons, a significant reduction in the number of cells exhibiting a response to α,β-meATP was observed. However, a subset of small diameter neurons retained P2X₃ responses of equal amplitude to those recorded from naive and sham control DRG neurons. Interestingly, P2X₃ immunoreactivity and P2X₃-like responses were also detected in a subset of larger diameter (50 μm) neurons and the number and amplitude of these responses were unchanged after spinal nerve ligation. These results suggest that, while there appears to be a decrease in fast desensitizing P2X₃ receptors following L5/L6 nerve ligation injury, certain subsets of small and large DRG neurons maintain normal P2X₃ receptor expression and function. These remaining receptors may provide a P2X₃ receptor-mediated component to neuropathic pain. Electronic Publication     

雪旺细胞源神经营养因子对背根节感觉神经元的保护作用

目的：了解雪旺细胞源神经营养因子对周围神经高位损伤所致脊髓背根节感觉神经元死亡的保护作用。方法：选出生3周SD鼠高位切断L4、L5神经根，神经近侧断端应用雪旺细胞源神经营养因子或生理盐水，4周后观察损伤神经根背根节感觉神经元的存活率和形态学变化。结果：术后4周，营养因子组神经元的存活率是91．8％，生理盐水组是58．6％；生理盐水组存活神经元胞体明显萎缩。结论：雪旺细胞源神经营养因子对受损的背根节感觉神经元有明显的神经营养活性．     

Enhancement of ATP-activated current by protons in dorsal root ganglion neurons

 The effect of pH on ATP-activated current in bullfrog dorsal root ganglion neurons was studied using the whole-cell patch-clamp technique. ATP-activated current amplitude was highly dependent upon extracellular pH. An acid pH increased, whereas alkaline pH decreased, ATP-activated current amplitude. The half-maximal pH (EC₅₀) for potentiation of 2.5 μM ATP-activated current was 7.2. Acidification alone did not activate detectable current and, at an acid pH, ATP-activated current was abolished by suramin. Proton-induced enhancement of ATP-activated current was not sensitive to membrane potential between –80 and +40 mV, and did not involve a shift in reversal potential. Lowering pH from 7.2 to 6.5 or elevating pH from 7.2 to 8.0 shifted the ATP concentration/response curve to the left or right, respectively, without changing the maximal response to ATP. Protons increased the time constant of deactivation without affecting the time constant of activation or desensitization of ATP-activated current. Alteration of patch-pipette (intracellular) pH did not affect the enhancement of ATP-activated current by extracellular protons. Diethylpyrocarbonate (DEP), dithiothreitol (DTT), 5,5′-dithio-bis-(2-nitro-benzoic acid) (DTNB), or N-ethylmaleimide (NEM) did not affect enhancement of ATP-activated current by protons. The results suggest that extracellular protons, at physiological concentrations, can regulate the function of P_2X purinoceptors by modulating the affinity of the ATP-binding site. Received: 20 July 1996 / Received after revision and accepted: 24 September 1996     

Innervation of the sacroiliac joint in rats by calcitonin gene-related peptide-immunoreactive nerve fibers and dorsal root ganglion neurons

The sacroiliac joint (SIJ) can be a source of low back pain. Calcitonin gene-related peptide (CGRP) has been reported to play a significant role in nociceptive processing. However, the occurrence of CGRP-immunoreactive (CGRP-ir) sensory nerve fibers in the SIJ has not been fully defined. The present study investigated CGRP-ir nerve fibers supplying the SIJ. CGRP-ir nerve fibers in the vicinity of the SIJ cartilage and CGRP-ir neurons in the bilateral dorsal root ganglia (DRG) were examined immunohistochemically by administering anti-CGRP antiserum to rats. The SIJ was decalcified and cut into sections, and the CGRP-ir fibers around the SIJ cartilage were counted under microscopy. In another group, fluoro-gold (F-G), a neural tracer, was injected into the SIJ from the dorsal or ventral side with dorsal or ventral denervation. The number of F-G-labeled CGRP-ir neurons was counted in individual DRG. CGRP-ir fibers were observed more frequently in the tissues adjacent to the cranial part of the SIJ surface. In the case of dorsal denervation (ventral nerve supply), the CGRP-ir neurons composed 18.2% of the F-G-labeled neurons. In the case of ventral denervation (dorsal nerve supply), the CGRP-ir neurons composed 40.9% of the F-G-labeled neurons. There was a statistically significant difference in the number of CGRP-ir neurons between the ventral and dorsal nerve supplies to the SIJ. The cranial part of the dorsal side could be the part most associated with pain in the SIJ.     

大麻素抑制背根节神经元ATP诱发的[Ca~(2+)]i升高

目的:观察大麻素对背根节神经元ATP诱发的[Ca~(2+)]i升高的影响及机制。方法:培养SD大鼠背根节神经元,采用激光共聚焦技术检测培养神经元[Ca~(2+)]i的变化。结果:ATP(100μmol/L)经P2X受体介导可导致培养的背根节神经元[Ca~(2+)]i增高(P0.05);大麻素受体激动剂CP55940预孵育10 min可剂量依赖性地抑制背根节神经元ATP所致的[Ca~(2+)]i升高(P0.05);CB1受体(cannabinoid receptor 1,B1R)的拮抗剂AM251(10μmol/L)、CB2受体(Cannabinoid receptor 2,CB2R)的拮抗剂AM630(10μmol/L)均可显著降低CP55940(1μmol/L)的抑制效应(P0.05);腺苷酸环化酶激动剂Forskolin(10μmol/L)可逆转CP55940对ATP的抑制作用(P0.05)。结论:CP55940可显著抑制背根节神经元ATP诱发的[Ca~(2+)]i升高,CP55940的抑制效应可能是由CB1、CB2受体介导抑制背根节神经元PKA活性所致。