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学习过程中蜗牛神经元基因组活性和突触可塑性的体内研究。

In vivo investigation of genome activity and synaptic plasticity of neurons in snails during learning.

作者信息

Shevelkin A V, Kozyrev S A, Nikitin V P, Sherstnev V V

机构信息

K. P. Anokhin Science Research Institute of Normal Physiology, Russian Academy of Medical Sciences, Building 4, 11 Mokhovaya Street, 103009 Moscow, Russia.

出版信息

Neurosci Behav Physiol. 2005 Jul;35(6):595-603. doi: 10.1007/s11055-005-0099-9.

Abstract

The present report describes in vivo investigations of genome activity and its role in the mechanisms forming long-term synaptic plasticity in defensive behavior command neuron LPl1 during the acquisition of nociceptive sensitization by common snails. Transcription processes were recorded using SYTO 16, a specific fluorescent indicator of DNA activity, along with in vivo computer image analysis. Studies in control snails showed that application of nociceptive stimuli to the head led to biphasic changes in the bioelectrical responses of neurons to tactile and chemical stimulation--depression of responses in the short-term stage (during the 1 h after training) and their facilitation during the long-term stage of sensitization (more than 24 h). There were marked increases in fluorescence over the nucleus of the command neuron stained with SYTO 16 at 15-20 min from the start of training, this lasting 4-5 h. Acquisition of sensitization in the presence of the RNA synthesis inhibitor actinomycin D (20 microM) to the neuron led to the complete elimination of changes in fluorescence and synaptic facilitation in the responses of LPl1 to sensory stimulation in the long-term stage of sensitization but had no effect during the short-term stage of sensitization. Actinomycin D given 30 min after the end of acquisition of sensitization (1 h after the start) had no effect on the dynamics of fluorescence or synaptic facilitation. Thus, the acquisition of nociceptive sensitization is accompanied by a rapid (within 15-20 min) activation of the DNA of neuron LPl1 and subsequent (about 1 h) display of long-term synaptic facilitation. Induction of both processes was suppressed by the RNA synthesis inhibitor over a relatively short time period--1 h from the moment at which training started.

摘要

本报告描述了在普通蜗牛获得伤害性敏感化过程中,对防御行为指令神经元LPl1的基因组活性及其在形成长期突触可塑性机制中的作用进行的体内研究。使用SYTO 16(一种DNA活性的特异性荧光指示剂)以及体内计算机图像分析记录转录过程。对对照蜗牛的研究表明,对头部施加伤害性刺激会导致神经元对触觉和化学刺激的生物电反应出现双相变化——在短期阶段(训练后1小时内)反应抑制,在敏感化的长期阶段(超过24小时)反应增强。从训练开始15 - 20分钟时,用SYTO 16染色的指令神经元细胞核上的荧光显著增加,持续4 - 5小时。在存在RNA合成抑制剂放线菌素D(20 microM)的情况下,神经元获得敏感化会导致在敏感化长期阶段LPl1对感觉刺激的反应中荧光变化和突触增强完全消除,但在敏感化短期阶段没有影响。在敏感化获得结束后30分钟(开始后1小时)给予放线菌素D对荧光动力学或突触增强没有影响。因此,伤害性敏感化的获得伴随着神经元LPl1的DNA迅速(15 - 20分钟内)激活以及随后(约1小时)长期突触增强的表现。RNA合成抑制剂在相对较短的时间段——从训练开始1小时内抑制了这两个过程的诱导。

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