研究报告

成年大脑神经细胞特异性ADAM10基因敲除小鼠模型的建立与鉴定

展开
  • 福建医科大学基础医学院细胞生物学与遗传学系, 细胞与发育工程研究中心, 干细胞工程与再生医学福建省高校重点实验室, 福州 350004

收稿日期: 2012-03-26

  修回日期: 2012-05-03

  网络出版日期: 2012-12-25

基金资助

null

Generation and characterization of the adult neuron-specific ADAM10 knock-out mice

Expand
  • Department of Cell Biology and Genetics, School of Basic Medicine / Center of Cell and Developmental Biology/ Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Fujian Medical University, Fuzhou 350004, China

Received date: 2012-03-26

  Revised date: 2012-05-03

  Online published: 2012-12-25

摘要

去整合素和金属蛋白酶10(ADAM10)是一种能够水解30余种跨膜蛋白质的“脱落酶”(sheddase), 参与诸多生理过程和致病机制, 如胚胎发育、细胞粘附、信号转导、免疫反应、癌症和阿尔茨海默病。迄今, 已报道的ADAM10完全基因敲除小鼠和大脑神经前体细胞特异性ADAM10基因敲除小鼠分别于胚胎期或围产期死亡, 致使无法研究成年小鼠大脑神经细胞ADAM10基因的功能。文章利用本研究小组建立的CaMKIIα-Cre转基因小鼠与ADAM10loxP/loxP转基因小鼠杂交, 获得了CaMKIIα-Cre/ADAM10loxP/loxP小鼠, 并对其进行鉴定。利用PCR方法检测成年ADAM10 cKO小鼠大脑基因组DNA表明, ADAM10基因缺失主要发生在前脑皮层和海马中。荧光定量PCR检测结果显示, ADAM10 mRNA的表达水平在前脑皮层和海马中分别降低55.7%和60.8% ; 使用Western blotting方法研究发现, ADAM10成熟蛋白质的含量在前脑皮层和海马中分别减少63%和84.8% 。采用免疫组织化学方法检测表明, 成年ADAM10 cKO小鼠与野生型小鼠相比, 其大脑皮层和海马神经细胞的ADAM10免疫染色明显减弱, 而其它细胞如胶质细胞的免疫染色基本一致。总之, 文章成功制备了首个存活至成年的大脑神经细胞特异性ADAM10基因敲除(cKO)小鼠, 克服了小鼠因ADAM10缺失在胚胎期或围产期死亡的弊端, 为研究成年小鼠大脑神经细胞ADAM10基因的功能奠定了坚实的基础。

本文引用格式

刘军,周常文,韦秋兰,庄建龙,林炤华,郑杰辉 . 成年大脑神经细胞特异性ADAM10基因敲除小鼠模型的建立与鉴定[J]. 遗传, 2012 , 34(12) : 1570 -1576 . DOI: 10.3724/SP.J.1005.2012.01570

参考文献

[1] Pruessmeyer J, Ludwig A. The good, the bad and the ugly substrates for ADAM10 and ADAM17 in brain pathology, inflammation and cancer. Semin Cell Dev Biol, 2009, 20(2): 164-174.
[2] Lichtenthaler SF. Alpha-secretase in Alzheimer's disease: molecular identity, regulation and therapeutic potential. J Neurochem, 2011, 116(1): 10-21.
[3] Kuhn PH, Wang HH, Dislich B, Colombo A, Zeitschel U, Ellwart JW, Kremmer E, Roβner S, Lichtenthaler SF. ADAM10 is the physiologically relevant, constitutive α-secretase of the amyloid precursor protein in primary neurons. EMBO J, 2010, 29(17): 3020-3032.
[4] Jorissen E, Prox J, Bernreuther C, Weber S, Schwanbeck R, Serneels L, Snellinx A, Craessaerts K, Thathiah A, Tesseur I, Bartsch U, Weskamp G, Blobel CP, Glatzel M, De Strooper B, Saftig P. The disintegrin/metalloproteinase ADAM10 is essential for the establishment of the brain cortex. J Neurosci, 2010, 30(14): 4833-4844.
[5] Kim M, Suh J, Romano D, Truong MH, Mullin K, Hooli B, Norton D, Tesco G, Elliott K, Wagner SL, Moir RD, Becker KD, Tanzi RE. Potential late-onset Alzheimer's disease-associated mutations in the ADAM10 gene attenuate α-secretase activity. Hum Mol Genet, 2009, 18(20): 3987- 3996.
[6] Prinzen C, Trümbach D, Wurst W, Endres K, Postina R, Fahrenholz F. Differential gene expression in ADAM10 and mutant ADAM10 transgenic mice. BMC Genomics, 2009, 10: 66.
[7] Parkin E, Harris B. A disintegrin and metalloproteinase (ADAM)-mediated ectodomain shedding of ADAM10. J Neurochem, 2009, 108(6): 1464-1479.
[8] Tousseyn T, Thathiah A, Jorissen E, Raemaekers T, Konietzko U, Reiss K, Maes E, Snellinx A, Serneels L, Nyabi O, Annaert W, Saftig P, Hartmann D, De Strooper B. ADAM10, the rate-limiting protease of regulated intramembrane proteolysis of Notch and other proteins, is processed by ADAMS-9, ADAMS-15, and the gamma-secretase. J Biol Chem, 2009, 284(17): 11738-11747.
[9] 尹海芳, 李宁, 王秋菊. 基因敲除鼠疾病模型的研究进展. 遗传, 2002, 24(4): 463-469.
[10] Altmeppen HC, Prox J, Puig B, Kluth MA, Bernreuther C, Thurm D, Jorissen E, Petrowitz B, Bartsch U, de Strooper B, Saftig P, Glatzel M. Lack of a-disintegrin-and-metalloproteinase ADAM10 leads to intracellular accumulation and loss of shedding of the cellular prion protein in vivo. BMC Mol Neurodegener, 2011, 6: 36.
[11] Weber S, Niessen MT, Prox J, Lüllmann-Rauch R, Schmitz A, Schwanbeck R, Blobel CP, Jorissen E, de Strooper B, Niessen CM, Saftig P. The disintegrin/metalloproteinase Adam10 is essential for epidermal integrity and Notch-mediated signaling. Development, 2011, 138(3): 495-505.
[12] Zhang C, Tian L, Chi CW, Wu XH, Yang X, Han M, Xu T, Zhuang Y, Deng KJ. Adam10 is essential for early embryonic cardiovascular development. Dev Dyn, 2010, 239(10): 2594-2602.
[13] Glomski K, Monette S, Manova K, de Strooper B, Saftig P, Blobel CP. Deletion of Adam10 in endothelial cells leads to defects in organ-specific vascular structures. Blood, 2011, 118(4): 1163-1174.
[14] Gibb DR, El Shikh M, Kang DJ, Rowe WJ, El Sayed R, Cichy J, Yagita H, Tew JG, Dempsey PJ, Crawford HC, Conrad DH. ADAM10 is essential for Notch2-dependent marginal zone B cell development and CD23 cleavage in vivo. J Exp Med, 2010, 207(3): 623-635.
[15] Tian L, Wu XH, Chi CW, Han M, Xu T, Zhuang Y. ADAM10 is essential for proteolytic activation of Notch during thymocyte development. Int Immunol, 2008, 20(9): 1181-1187.
[16] Hartmann D, de Strooper B, Serneels L, Craessaerts K, Herreman A, Annaert W, Umans L, Lübke T, Lena Illert A, von Figura K, Saftig P. The disintegrin/metalloprotease ADAM 10 is essential for Notch signalling but not for alpha-secretase activity in fibroblasts. Hum Mol Genet, 2002, 11(21): 2615-2624.
[17] 徐瑛, 周常文, 林炤华, 柯琦. CaMKⅡα基因启动子的克隆和神经细胞特异性Cre表达载体的构建. 中国优生与遗传杂志, 2008, 1
文章导航

/