腺苷对脊髓背角NMDA受体的抑制性调节作用
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摘要
目的研究腺苷对脊髓背角N-甲基-D-天冬氨酸(NMDA)型谷氨酸受体的调节作用。方法制备SD大鼠离体脊髓片,在脊髓背角Ⅱ板层神经元上膜片钳全细胞记录NMDA受体介导的兴奋性突触后电流(EPSCs);提取脊髓背角突触后致密质(PSD)结构,蛋白质印迹法检查NMDA受体的含量。结果胞外灌流腺苷(50μmol/L)能够明显降低EPSCs的幅值(P<0.05),加速EPSCs的衰减(P<0.05);能够显著降低Glu N2B受体在兴奋性突触中的含量(P<0.05)。结论腺苷可能通过降低Glu N2B受体在突触的表达水平,抑制NMDA受体的痛觉突触传递。
Objective To investigate the regulatory effect of adenosine on N-methyl-D-aspartate(NMDA)subtype glutamate receptor in spinal cord dorsal horn. Methods The spinal cord slices were prepared from SD rats. The excitatory postsynaptic currents(EPSCs) mediated by NMDA receptors were recorded in lamina Ⅱneurons under whole-cell patch clamp configuration. Postsynaptic density(PSD) was fractionated and immunoblotted to analyze the synaptic expression of NMDA receptor. Results Extracellular perfusion of adenosine(50 μmol/L) significantly decreaseds the amplitude of EPSCs and accelerated the current decay(P < 0.05).Meanwhile, adenosine noticeably reduced the synaptic contents Glu N2 B receptor(P < 0.05). Conclusion Adenosine might blunt NMDA receptor-mediated nociceptive transmission by removing Glu N2 B receptor out of synapses.
Objective To investigate the regulatory effect of adenosine on N-methyl-D-aspartate(NMDA)subtype glutamate receptor in spinal cord dorsal horn. Methods The spinal cord slices were prepared from SD rats. The excitatory postsynaptic currents(EPSCs) mediated by NMDA receptors were recorded in lamina Ⅱneurons under whole-cell patch clamp configuration. Postsynaptic density(PSD) was fractionated and immunoblotted to analyze the synaptic expression of NMDA receptor. Results Extracellular perfusion of adenosine(50 μmol/L) significantly decreaseds the amplitude of EPSCs and accelerated the current decay(P < 0.05).Meanwhile, adenosine noticeably reduced the synaptic contents Glu N2 B receptor(P < 0.05). Conclusion Adenosine might blunt NMDA receptor-mediated nociceptive transmission by removing Glu N2 B receptor out of synapses.
引文
[1]冷玉芳,金海燕.大鼠坐骨神经慢性压迫性损伤后脊髓背角感觉神经元超微形态学变化[J].兰州大学学报:医学版,2009,35(4):31-34.
[2]曹晓曼,张胜男,张福康,等.经典瞬时感受器电位通道在炎性痛及神经病理性痛大鼠背根神经节表达水平的差异[J].兰州大学学报:医学版,2014,40(3):1-7.
[3]Zylka M J.Pain-relieving prospects for adenosine receptors and ectonucleotidases[J].Trends Mol Med,2011,17(4):188-196.
[4]Lau C G,Zukin R S.NMDA receptor trafficking in synaptic plasticity and neuropsychiatric disorders[J].Nat Rev Neurosci,2007,8(6):413-426.
[5]Chizh B A,Reissmüller E,Schlütz H,et al.Supraspinal vs spinal sites of the antinociceptive action of the subtypeselective NMDA antagonist ifenprodil[J].Neuropharmacology,2001,40(2):212-220.
[6]Zhang W B,Ross P J,Tu Y,et al.Fyn kinase regulates Glu N2B subunit-dominant NMDA receptors in human induced pluripotent stem cell-derived neurons[J].Sci Rep,2016,6:238-237.
[7]Wang,W T,Pan G Q,Zang Z Y,et al.Ht31 peptide inhibited inflammatory pain by blocking NMDA receptormediated nociceptive transmission in spinal dorsal horn of mice[J].Neuropharmacology,2015,89:290-297.
[8]Hu X D,Liu Y N,Zhang Z Y,et al.Spinophilin-targeted protein phosphatase-1 alleviated inflammatory pain by negative control of MEK/ERK signaling in spinal cord dorsal horn of rats[J].J Neurosci,2015,35(41):13989-14001.
[9]Yang X,Yang H B,Xie Q J,et al.Peripheral inflammation increased the synaptic expression of NMDA receptors in spinal dorsal horn[J].Pain,2009,144(1-2):162-169.
[10]Smith K E,Gibson E S,Dell'Acqua M L.c AMP-dependent protein kinase postsynaptic localization regulated by NMDA receptor activation through translocation of an Akinase anchoring protein scaffold protein[J].J Neurosci,2006,26(9):2391-2402.
[11]Jaworski J,Kapitein L C,Gouveia S M,et al.Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity[J].Neuron,2009,61(1):85-100.
[12]Sanz-Clemente A,Matta J A,Isaac J T,et al.Casein kinase 2 regulates the NR2 subunit composition of synaptic NMDA receptors[J].Neuron,2010,67(6):984-996.
[13]Sawynok J.Adenosine receptor targets for pain[J].Neuroscience,2016,338:1-18.
[14]Tao Y X,Rumbaugh G,Wang G D,et al.Impaired NMDA receptor-mediated postsynaptic function and blunted NMDA receptor-dependent persistent pain in mice lacking postsynaptic density-93 protein[J].J Neurosci,2003,23(17):6703-6712.
[15]Liu X,Chen C.Different roles for AMPA and NMDA receptors in transmission at the immature retinogeniculate synapse[J].J Neurophysiol,2008,99(2):629-643.
[16]Wang C C,Held R G,Chang S C,et al.A critical role for Glu N2B-containing NMDA receptors in cortical development and function[J].Neuron,2011,72(5):789-805.
[17]Nascimento F P,Macedo-Júnior S J,Pamplona F A,et al.Adenosine A1 receptor-dependent antinociception induced by inosine in mice:pharmacological,genetic and biochemical aspects[J].Mol Neurobiol,2015,51(3):1368-1378.
[18]Sanz-Clemente A,Nicoll R A,Roche K W.Diversity in NMDA receptor composition:many regulators,many consequences[J].Neuroscientist,2013,19(1):62-75.
[19]Barth A L,Malenka R C.NMDAR EPSC kinetics do not regulate the critical period for LTP at thalamocortical synapses[J].Nat Neurosci,2001,4(3):235-236.
[20]Wu L J,Zhuo M.Targeting the NMDA receptor subunit NR2B for the treatment of neuropathic pain[J].Neurotherapeutics,2009,6(4):693-702.
[21]Zhuo M.Plasticity of NMDA receptor NR2B subunit in memory and chronic pain[J].Mol Brain,2009,2:4.
[2]曹晓曼,张胜男,张福康,等.经典瞬时感受器电位通道在炎性痛及神经病理性痛大鼠背根神经节表达水平的差异[J].兰州大学学报:医学版,2014,40(3):1-7.
[3]Zylka M J.Pain-relieving prospects for adenosine receptors and ectonucleotidases[J].Trends Mol Med,2011,17(4):188-196.
[4]Lau C G,Zukin R S.NMDA receptor trafficking in synaptic plasticity and neuropsychiatric disorders[J].Nat Rev Neurosci,2007,8(6):413-426.
[5]Chizh B A,Reissmüller E,Schlütz H,et al.Supraspinal vs spinal sites of the antinociceptive action of the subtypeselective NMDA antagonist ifenprodil[J].Neuropharmacology,2001,40(2):212-220.
[6]Zhang W B,Ross P J,Tu Y,et al.Fyn kinase regulates Glu N2B subunit-dominant NMDA receptors in human induced pluripotent stem cell-derived neurons[J].Sci Rep,2016,6:238-237.
[7]Wang,W T,Pan G Q,Zang Z Y,et al.Ht31 peptide inhibited inflammatory pain by blocking NMDA receptormediated nociceptive transmission in spinal dorsal horn of mice[J].Neuropharmacology,2015,89:290-297.
[8]Hu X D,Liu Y N,Zhang Z Y,et al.Spinophilin-targeted protein phosphatase-1 alleviated inflammatory pain by negative control of MEK/ERK signaling in spinal cord dorsal horn of rats[J].J Neurosci,2015,35(41):13989-14001.
[9]Yang X,Yang H B,Xie Q J,et al.Peripheral inflammation increased the synaptic expression of NMDA receptors in spinal dorsal horn[J].Pain,2009,144(1-2):162-169.
[10]Smith K E,Gibson E S,Dell'Acqua M L.c AMP-dependent protein kinase postsynaptic localization regulated by NMDA receptor activation through translocation of an Akinase anchoring protein scaffold protein[J].J Neurosci,2006,26(9):2391-2402.
[11]Jaworski J,Kapitein L C,Gouveia S M,et al.Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity[J].Neuron,2009,61(1):85-100.
[12]Sanz-Clemente A,Matta J A,Isaac J T,et al.Casein kinase 2 regulates the NR2 subunit composition of synaptic NMDA receptors[J].Neuron,2010,67(6):984-996.
[13]Sawynok J.Adenosine receptor targets for pain[J].Neuroscience,2016,338:1-18.
[14]Tao Y X,Rumbaugh G,Wang G D,et al.Impaired NMDA receptor-mediated postsynaptic function and blunted NMDA receptor-dependent persistent pain in mice lacking postsynaptic density-93 protein[J].J Neurosci,2003,23(17):6703-6712.
[15]Liu X,Chen C.Different roles for AMPA and NMDA receptors in transmission at the immature retinogeniculate synapse[J].J Neurophysiol,2008,99(2):629-643.
[16]Wang C C,Held R G,Chang S C,et al.A critical role for Glu N2B-containing NMDA receptors in cortical development and function[J].Neuron,2011,72(5):789-805.
[17]Nascimento F P,Macedo-Júnior S J,Pamplona F A,et al.Adenosine A1 receptor-dependent antinociception induced by inosine in mice:pharmacological,genetic and biochemical aspects[J].Mol Neurobiol,2015,51(3):1368-1378.
[18]Sanz-Clemente A,Nicoll R A,Roche K W.Diversity in NMDA receptor composition:many regulators,many consequences[J].Neuroscientist,2013,19(1):62-75.
[19]Barth A L,Malenka R C.NMDAR EPSC kinetics do not regulate the critical period for LTP at thalamocortical synapses[J].Nat Neurosci,2001,4(3):235-236.
[20]Wu L J,Zhuo M.Targeting the NMDA receptor subunit NR2B for the treatment of neuropathic pain[J].Neurotherapeutics,2009,6(4):693-702.
[21]Zhuo M.Plasticity of NMDA receptor NR2B subunit in memory and chronic pain[J].Mol Brain,2009,2:4.