亲代谢型谷氨酸受体5在Homer 1a RNA干扰大鼠学习记忆障碍中的作用

中国儿童保健杂志 ›› 2013, Vol. 21 ›› Issue (1): 50-53.

亲代谢型谷氨酸受体5在Homer 1a RNA干扰大鼠学习记忆障碍中的作用

洪琴¹,池霞^1,2,杨蕾²,郭锡熔^1,2,童梅玲^1,2,张敏²,陈荣华²,费莉²,郭梅²,潘晓勤²

1 南京医科大学附属南京妇幼保健院儿保科,江苏南京 210004;
2 南京医科大学儿科医学研究所,江苏南京 210029

收稿日期:2012-09-27 发布日期:2013-01-06 出版日期:2013-01-06
通讯作者: 池霞,E-mail:chixia2001@yahoo.com.cn
作者简介:洪琴(1982-),女,主治医师,博士在读,主要研究方向为发育行为儿科学。
【通信作者】池霞,E-mail:chixia2001@yahoo.com.cn
基金资助:
国家自然科学基金(30801255);国家自然科学基金(81250008);江苏省医学重点人才(RC2011022)

Effect of metabotropic glutamate receptor 5 on learning memory of Homer 1a RNA interference rats

HONG Qin¹,CHI Xia^1.2,YANG Lei²,GUO Xi-rong^1.2,TONG Mei-ling¹,ZHANG Min²,CHEN Rong-hua¹,FEI Li¹,GUO Mei¹,PAN Xiao-qin¹.

1 Children Health Care Center,Nanjing Maternal and Children Health Hospital of Nanjing Medical University,Nanjing,Jiangsu 210004,China;
2 Institute of Pediatrics,Nanjing Medical University,Nanjing,Jiangsu 210029,China

Received:2012-09-27 Online:2013-01-06 Published:2013-01-06

摘要/Abstract

摘要： 目的 Homer 1a RNA干扰(RNA interference,RNAi)后的Sprague Dawley (SD)大鼠在Morris水迷宫中表现学习记忆能力障碍,本研究旨在通过检测Homer 1a RNAi后大鼠不同脑区亲代谢型谷氨酸受体5(metabotropic glutamate receptor 5,mGluR 5)的表达情况,来初步探讨Homer 1a RNAi致大鼠学习记忆障碍的分子机制。方法将24只SD大鼠随机分为两组: Homer 1a RNAi组(RNAi组,n=12)和对照组(Nc组,n=12),运用侧脑室注射技术,分别向两组大鼠的侧脑室注射针对Homer 1a 的RNAi慢病毒和无义病毒,等待病毒起效时间7 d,完成相应的行为学检测(Morris水迷宫)后,利用荧光定量PCR 和Western blot 技术检测不同脑组织(前额叶、纹状体、海马)mGluR5 mRNA 和蛋白表达情况。结果 RNAi组大鼠纹状体和海马mGluR5 mRNA和蛋白表达均较对照组显著降低(纹状体:mRNA t=6.67,蛋白t=3.10;海马:mRNA t=3.08;蛋白t=2.89,P值均<0.05或<0.01),前额叶mGluR5 mRNA和蛋白表达较对照组差异无统计学意义(mRNA t=1.58,蛋白t=1.66,P值均>0.05)。结论 Homer 1a RNAi后大鼠脑组织Homer 1a相关基因mGluR5表达下调,提示mGluR5可能作为Homer 1a的下游基因,参与Homer 1a RNAi后大鼠学习记忆的损伤过程。

关键词: RNA干扰, ADHD, Homer 1a, 亲代谢型谷氨酸受体5, 学习记忆

Abstract: Objective Learning and memory abilities of rats were observed damaged in the Morris water maze,after expressions of Homer 1a in their brain interfered by RNA interference (RNA interference,RNAi).Expressions of mGluR 5 in their brain were detected to explore the molecular mechanism of learning and memory impairment after expressions of Homer 1a in their brain interfered by RNAi. Methods 24 Sprague Dawley (SD) rats were randomly divided into two groups:Homer 1a RNAi group (RNAi group,n=12) and control group (Nc group,n=12),and injected into the intracerebroventricular of the rats with Homer 1a RNAi lentivirus or nonsense lentivirus,respectively.Seven days later,completed the behavior tests (Morris Water Maze),mRNA and protein expressions of mGluR5 in some regions (prefrontal cortex,striatum,hippocampus) were analyzed using Quantitative PCR and Western blot. Result mGluR5 mRNA and protein expression in the striatum and hippocampus of RNAi group compared with the control group were significantly lower (striatum:mRNA t=6.67,protein t=3.10;hippocampus:mRNA t=3.08;protein t=2.89,all P<0.05 or <0.01),while there were no significant differences in the prefrontal(mRNA t=1.58,protein t=1.66,all P>0.05). Conclusion Homer 1a RNAi rats exhibit learning and memory impairment,and the expressions of Homer 1a and its related gene mGluR5 are changed,suggesting that mGluR5,as Homer 1a downstream gene,is involved in the mechanisms of learning and memory impairment after expressions of Homer 1a in their brain interfered by RNAi.

Key words: RNA interference, ADHD, Homer 1a, metabotropic glutamate receptor 5, learning and memory

中图分类号:

Q95-33

洪琴,池霞,杨蕾,郭锡熔,童梅玲,张敏,陈荣华,费莉,郭梅,潘晓勤. 亲代谢型谷氨酸受体5在Homer 1a RNA干扰大鼠学习记忆障碍中的作用[J]. 中国儿童保健杂志, 2013, 21(1): 50-53.

HONG Qin,CHI Xia,YANG Lei,GUO Xi-rong,TONG Mei-ling,ZHANG Min,CHEN Rong-hua,FEI Li,GUO Mei,PAN Xiao-qin.. Effect of metabotropic glutamate receptor 5 on learning memory of Homer 1a RNA interference rats[J]. journal1, 2013, 21(1): 50-53.

参考文献

[1] 张军峰,刘勇.代谢型谷氨酸受体与神经发生的关系[J].医学综述,2009,15(22):3386-3388.
[2] Sokol DK,Maloney B,Long JM,et al.Autism,alzheimer disease,and fragile X:APP,FMRP,and mGluR5 are molecular links[J].Neurology,2011,76(15):1344-1352.
[3] Enz R.Metabotropic glutamate receptors and interacting proteins:evolving drug targets[J].Curr Drug Targets,2012,13(1):145-156.
[4] Grassi S,Frondaroli A,Pettorossi VE.Role of group Ⅱmetabotropic glutamate receptors 2/3 and group Ⅰmetabotropic glutamate receptor 5 in developing rat medial vestibular nuclei[J].Neuroreport,2005,16(12):1303-1307.
[5] Koros E,Rosenbrock H,Birk G,et al.The selective mGlu5 receptor antagonist MTEP,similar to NMDA receptor antagonists,induces social isolation in rats[J].Neuropsychopharmacology,2007,32(3):562-576.
[6] Balschun D,Zuschratter W,Wetzel W.Allosteric enhancement of metabotropic glutamate receptor 5 function promotes spatial memory[J].Neuroscience,2006,142(3):691-602.
[7] Balschun D,Wetzel W.Inhibition of mGluR5 blocks hippocampal LTP in vivo and spatial learning in rats[J].Pharmacol Biochem Behav,2002,73(2):375-380.
[8] Manahan-Vaughan D,Braunewell KH.The metabotropic glutamate receptor,mGluR5,is a key determinant of good and bad spatial learning perfor mance and hippocampal synaptic plasticity[J].Cereb Cortex,2005,15(11):1703-1713.
[9] Tappe-Theodor A,Fu Y,Kuner R,et al.Homer1a signaling in the amygdala counteracts pain-related synaptic plasticity,mGluR1 function and pain behaviors[J].Mol Pain,2011,7(38):1-6.
[10] Feldman LA,Shapiro ML,Nalbantoglu J.A novel,rapidly acquired and persistent spatial memory task that induces immediate early gene expression[J].Behav Brain Funct,2010,6(35):1-11.
[11] Hu JH,Park JM,Park S,et al.Homeostatic scaling requires group I mGluR activation mediated by Homer1a[J].Neuron,2010,68(6):1128-1142.
[12] Gerstein H,O'Riordan K,Osting S,et al.Rescue of synaptic plasticity and spatial learning deficits in the hippocampus of Homer1 knockout mice by recombinant Adeno-associated viral gene delivery of Homer1c[J].Neurobiol Learn Mem,2012,97(1):17-29.
[13] Ango F,Pin JP,Tu JC,et al.Dendritic and axonal targeting of type 5 metabotropic glutamate receptor is regulated by homer1 proteins and neuronal excitation[J].J Neurosci,2000,20(23):8710-8716.
[14] Sala C,Roussignol G,Meldolesi J,et al.Key role of the postsynaptic density scaffold proteins Shank and Homer in the functional architecture of Ca²⁺ homeostasis at dendritic spines in hippocampal neurons[J].J Neurosci,2005,25(18):4587-4592.
[15] Ango F,Robbe D,Tu JC,et al.Homer dependent cell surface expression of metabotropic glutamate receptor type 5 in neurons[J].Mol Cell Neurosci,2002,20:323-329.
[16] Jia Z,Lu Y,Henderson J,et al.Selective abolition of the NMDA component of long term potentiation in mice lacking mGluR5[J].Learn Mem,1998,5(4):331-343.
[17] Ullman MT,Pierpont EI.Specific language impairment is not specific to language:the procedural deficit hypothesis[J].Cortex,2005,41(3):399-433.