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吴爽,斯易提,陈舒芸,等.neuroplastin 65敲除对小鼠学习与记忆功能的影响[J].同济大学学报（医学版）,2014,35(4):1-6. [点击复制]
WU Shuang,Si Yi-ti,CHEN Shu-yun,et al.Effects of neuroplastin 65 knockout on the learning ability and spatial memory of mice[J].同济大学学报（医学版）,2014,35(4):1-6. [点击复制]

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neuroplastin 65敲除对小鼠学习与记忆功能的影响
吴爽¹,斯易提²,陈舒芸³,李颖²,蒋灵芝¹,袁琼兰²
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(1.深圳大学生命科学学院,广东深圳528060;;2.同济大学医学院解剖学与神经生物学教研室,上海200092;;3.同济大学医学院2010级临床医学系,上海200092)

摘要:

目的研究neuroplastin 65（NP65）对小鼠学习与记忆功能的影响。方法利用基因打靶获得NP65（-/-）的C57BL/6小鼠。培养新生鼠的海马神经元48 h,观察神经元神经突起的生长及免疫染色观察MAP-2、NP65的表达;培养海马神经元96 h,观察神经元活性;成年小鼠脑组织切片,尼氏染色、MAP-2及GFAP的免疫染色观察神经元及星形胶质细胞。Morris水迷宫实验观察小鼠的学习能力与空间记忆的改变。结果培养的NP65（-/-）小鼠海马神经元仅表达MAP-2,不表达NP65;与野生型相比,NP65（-/-）小鼠的海马神经元的最长的突起变短但初级突起的数目增加。脑组织切片尼氏染色显示,NP65（-/-）小鼠海马神经元的密度比野生型大,NP65（-/-）小鼠海马区MAP-2表达较野生型增加;GFAP阳性的星形胶质细胞数量比野生型多。NP65（-/-）小鼠的海马神经元不表达NP65。水迷宫实验显示NP65（-/-）小鼠的逃逸时间比野生型短、穿越平台次数的比野生型多。结论NP65（-/-）影响海马神经元突起生长;NP65敲除提高了小鼠的学习与空间记忆能力。

关键词: NP65 敲除神经突起生长学习与记忆

DOI：10.3969/j.issn1008-0392.2014.04.001

基金项目:国家自然科学基金（81070987;81371213）; 上海市大学生创新活动计划（1500107066）

Effects of neuroplastin 65 knockout on the learning ability and spatial memory of mice

WU Shuang¹,Si Yi-ti²,CHEN Shu-yun³,LI Ying JIANG²,JIANG Ling-zhi¹,YUAN Qiong-lan²

(1. College of Life Science, Shenzhen University, Shenzhen 528060, Guangdong Province, China;;2. Dept. of Anatomy and Neurobiology, School of Medicine, Tongji University, Shanghai 200092, China;;3. Dept. of Clinical Medicine, School of Medicine, Tongji University, Shanghai 200092, China)

Abstract:

Objective To investigate the effect of neuroplastin 65 (NP65) on the learning ability and spatial memory of mice. Methods NP65 ( -/-）mice were derived from the offspring of NP65 (+/-) C57BL/6 mice. The hippocampal neurons from newborn NP65 ( -/- ) and wild type C57BL/6 mice were cultured for 48 h, then immunostained with NP65 and microtubule-associated protein-2 (MAP-2) antibodies. The neurite outgrowth was evaluated with phase contrast inverted microscope. Adult NP65 ( -/- ) and wild type mice were sacrificed and cerebral coronal slices were used for Nissl staining and immunostaining. Morris water maze test was used to evaluate hippocampus-dependent learning and memory function in adult mice. Results The cultured hippocampal neurons from NP65 ( -/- ) mice expressed only MAP-2 but not NP65. Compared with wild type mice，the number of primary neurites of NP65 (-/- ) mice was significantly increased，but the longest length of neurites of NP65 ( -/- ) mice were significantly reduced. Nissl staining showed that the density of hippocampal neurons of NP65 ( -/- ) mice was higher than that of wild type mice. Immunostaining demonstrated that expression of MAP-2 in hippocampus of NP65 ( -/- ) mice was increased compared with wild type mice, but there was negative staining of NP65 in NP65 ( -/- ) mice. GFAP positive cells were increased in NP65 ( -/- ) mice. Morris water maze test showed that adult NP65 ( -/- ) mice had shorter escape latency than that of wild type mice； probe test showed that the number of crossing over the previous platform of NP65 ( -/- ) mice was increased compared with wild type mice. Conclusion NP65 can affect neurite outgrowth of hippocampal neurons and improve learning ability and spatial memory in mice.

Key words: NP65 knockout neurite outgrowth learning and spatial memory