中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (5): 880-886.doi: 10.4103/1673-5374.268929

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

幼年氯胺酮暴露诱导大脑海马神经元凋亡可影响成年空间学习能力

  

  • 出版日期:2020-05-15 发布日期:2020-06-01
  • 基金资助:
    国家自然科学基金项目(81200851);美国国家卫生研究院项目(NS 040723)

Neonatal ketamine exposure-induced hippocampal neuroapoptosis in the developing brain impairs adult spatial learning ability

Dan Lyu1, 2, Ning Tang2, 3, Andrew W. Womack2, Yong-Jin He1, Qing Lin2   

  1. 1 Department of Pain Management, Tianjin First Center Hospital, Tianjin, China
    2 Department of Psychology, College of Science, University of Texas at Arlington, Arlington, TX, USA
    3 Reproductive Medicine Center, the 960th Hospital of the PLA Joint Logistics Support Force, Jinan, Shandong Province, China
  • Online:2020-05-15 Published:2020-06-01
  • Contact: Qing Lin, MD, PhD,qilin@uta.edu; Yong-Jin He, MM,heyongjin1995@sina.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 81200851 (to DL); the National Institutes of Health of the USA, No. NS 040723 (to QL).

摘要:

氯胺酮暴露可选择性诱导发育中大脑神经细胞的凋亡。p66ShcA是一种在未成熟神经元中选择性表达的细胞衔接蛋白,也是一种促细胞凋亡分子,在激活时可诱发神经细胞凋亡。(1)实验首先对出生后7d的SD大鼠进行20mg/kg氯胺酮颈部皮下注射,每隔2h一次,共6次。注射氯胺酮结束后0,1,3,6h,以Western blot检测cleaved caspase-3,p66ShcA以及磷酸化p66ShcA的表达;(2)结果发现氯胺酮暴露后,激活的p66ShcA和caspase-3的表达增加,且在3h时达到峰值;(3)将出生后7d大鼠分别以20mg/kg氯胺酮皮下注射6次,每隔2h一次,于4周龄时通过Morris水迷宫测试空间学习和记忆能力。发现与幼时未进行氯胺酮干预的大鼠相比,这些大鼠的寻找隐藏平台的潜伏期延长,但在目标象限的停留时间接近;(4)实验结果表明,p66ShcA在新生儿期氯胺酮暴露诱导的凋亡级联过程被激活,且氯胺酮导致的空间学习缺陷可持续到成年期。实验于2013年1月22日经美国德克萨斯大学阿灵顿分校动物护理和使用委员会批准,批准号A13.008。

orcid: 0000-0002-7455-9937 (Qing Lin) 

         0000-0003-3579-670X (Yong-Jin He)

关键词: 氯胺酮, p66ShcA, 发育中的神经细胞凋亡, 海马, 空间学习能力, caspase-3, Morris水迷宫, N-甲基-D-天冬氨酸受体, 神经再生

Abstract: Ketamine exposure can lead to selective neuroapoptosis in the developing brain. p66ShcA, the cellular adapter protein expressed selectively in immature neurons, is a known pro-apoptotic molecule that triggers neuroapoptosis when activated. Sprague-Dawley rats at postnatal day 7 were subcutaneously injected in the neck with ketamine 20 mg/kg, six times at 2-hour intervals. At 0, 1, 3, and 6 hours after final injection, western blot assay was used to detect the expression of cleaved caspase-3, p66ShcA, and phosphorylated p66ShcA. We found that the expression of activated p66ShcA and caspase-3 increased after ketamine exposure and peaked at 3 hours. The same procedure was performed on a different group of rats. At the age of 4 weeks, spatial learning and memory abilities were tested with the Morris water maze. Latency to find the hidden platform for these rats was longer than it was for control rats, although the residence time in the target quadrant was similar. These findings indicate that ketamine exposure resulted in p66ShcA being activated in the course of an apoptotic cascade during the neonatal period. This may have contributed to the deficit in spatial learning and memory that persisted into adulthood. The experimental protocol was approved by the Institutional Animal Care and Use Committee at the University of Texas at Arlington, USA (approval No. A13.008) on January 22, 2013.

Key words: caspase-3, developmental neuroapoptosis, hippocampus, ketamine, Morris water maze, N-methyl-D-aspartate acid receptors, p66ShcA, spatial learning