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猫脊髓切片中交感神经节前神经元的快速兴奋性突触后电位及对兴奋性氨基酸的反应

Fast excitatory postsynaptic potentials and the responses to excitant amino acids of sympathetic preganglionic neurons in the slice of the cat spinal cord.

作者信息

Inokuchi H, Yoshimura M, Yamada S, Polosa C, Nishi S

机构信息

Department of Physiology, Kurume University School of Medicine, Japan.

出版信息

Neuroscience. 1992;46(3):657-67. doi: 10.1016/0306-4522(92)90152-r.

Abstract

The properties of the excitatory postsynaptic potential evoked by focal stimulation and of the responses to excitatory amino acids were examined by intracellular recording from sympathetic preganglionic neurons in upper thoracic spinal cord slices of the adult cat. Single stimuli to the region dorsal to the intermedio-lateral nucleus evoked short-latency, presumably monosynaptic, excitatory postsynaptic potentials. The reversal potential of this response was -2.2 mV and became more negative when external Na+ or K+ concentration was decreased. The excitatory postsynaptic potential was depressed by the non-selective excitatory amino acid receptor antagonist cis-2,3-piperidine dicarboxylic acid and enhanced by a glutamate uptake inhibitor. The non-N-methyl-D-aspartate receptor antagonist 6-cyano-7-nitroquinoxaline-2.3-dione abolished the excitatory postsynaptic potential in 72% of neurons. In the remaining neurons, this antagonist only depressed the potential and unmasked a slower component which was abolished by the N-methyl-D-aspartate receptor antagonist D,L-2-amino-5-phosphonovaleric acid. In the presence of tetrodotoxin all neurons tested were depolarized by glutamate or aspartate, as well as by the selective agonists quisqualate, alpha-amino-3-hydroxy-5-methylisoxazole propionic acid, kainate and N-methyl-D-aspartate. The glutamate-evoked depolarization reversed at a membrane potential of -2.0 mV and at a more negative value when external Na+ or K+ concentration was decreased. The response to alpha-amino-3-hydroxy-5-methylisoxazole propionic acid was abolished by 6-cyano-7-nitroquinoxaline-2,3-dione in all neurons tested and that to kainate in only one-third of the cells. In the remainder the response to kainate was only slightly depressed by this antagonist. The responses to glutamate and aspartate were only slightly depressed by the combined action of the various amino acid receptor antagonists used. The responses to N-methyl-D-aspartate were abolished by D,L-2-amino-5-phosphonovaleric acid. The punched-out region of the intermedio-lateral nucleus, maintained in vitro, released glutamate and aspartate in the absence of stimulation. Field stimulation (20 Hz) enhanced release by between 40 and 100%. The increase was prevented by superfusion with calcium-free Krebs. It is concluded that excitatory amino acids, acting on both N-methyl-D-aspartate and non-N-methyl-D-aspartate receptors, but mainly on the latter, are likely mediators of the monosynaptic excitatory postsynaptic potential evoked in sympathetic preganglionic neurons by the stimulated region. The efflux data suggest that glutamate and aspartate are among the mediators.

摘要

采用细胞内记录法,对成年猫上胸段脊髓切片中交感神经节前神经元进行研究,检测了局灶性刺激诱发的兴奋性突触后电位的特性以及对兴奋性氨基酸的反应。对中间外侧核背侧区域进行单次刺激可诱发潜伏期短的、推测为单突触的兴奋性突触后电位。该反应的反转电位为-2.2 mV,当细胞外Na+或K+浓度降低时,反转电位变得更负。非选择性兴奋性氨基酸受体拮抗剂顺式-2,3-哌啶二羧酸可抑制兴奋性突触后电位,而谷氨酸摄取抑制剂则可增强该电位。非N-甲基-D-天冬氨酸受体拮抗剂6-氰基-7-硝基喹喔啉-2,3-二酮可使72%的神经元的兴奋性突触后电位消失。在其余神经元中,该拮抗剂仅使电位降低,并暴露出一个较慢的成分,该成分可被N-甲基-D-天冬氨酸受体拮抗剂D,L-2-氨基-5-膦酰基戊酸所消除。在河豚毒素存在的情况下,所有受试神经元均被谷氨酸、天冬氨酸以及选择性激动剂quisqualate、α-氨基-3-羟基-5-甲基异恶唑丙酸、海人藻酸和N-甲基-D-天冬氨酸去极化。谷氨酸诱发的去极化在膜电位为-2.0 mV时反转,当细胞外Na+或K+浓度降低时,反转电位更负。在所有受试神经元中,6-氰基-7-硝基喹喔啉-2,3-二酮可消除对α-氨基-3-羟基-5-甲基异恶唑丙酸的反应,而仅在三分之一的细胞中可消除对海人藻酸的反应。在其余细胞中,该拮抗剂仅使对海人藻酸的反应略有降低。多种氨基酸受体拮抗剂联合作用仅使对谷氨酸和天冬氨酸的反应略有降低。D,L-2-氨基-5-膦酰基戊酸可消除对N-甲基-D-天冬氨酸的反应。在体外维持的中间外侧核的穿孔区域,在无刺激的情况下可释放谷氨酸和天冬氨酸。场刺激(20 Hz)可使释放增加40%至100%。用无钙的 Krebs 液灌注可阻止这种增加。由此得出结论,兴奋性氨基酸作用于N-甲基-D-天冬氨酸受体和非N-甲基-D-天冬氨酸受体,但主要作用于后者,它们可能是受刺激区域在交感神经节前神经元中诱发单突触兴奋性突触后电位的介质。外流数据表明,谷氨酸和天冬氨酸是介质之一。

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