Selection of genes associated with variations in the Circle of Willis in gerbils using suppression subtractive hybridization.
Selection of genes associated with variations in the Circle of Willis in gerbils using suppression subtractive hybridization.
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利用抑制消减杂交选择与沙鼠威利斯环变异相关的基因
DOI：
10.1371/journal.pone.0127355
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Chen Z
中科院分区：
文献类型：
--
作者：
Li Z;Huo X;Zhang S;Lu J;Li C;Guo M;Fu R;He Z;Du X;Chen Z
Willis环（CoW）畸形可显著增加人类脑血管疾病的风险。然而，这些畸形背后的分子机制尚未被理解。根据我们以前的研究，沙鼠的牛的变化是遗传性的。在大约60%的沙鼠中观察到正常的CoW，这一百分比也适用于人类。因此，沙鼠是一个理想的实验模型，研究在CoW的变化。为了研究这些变化的机制，我们选择了与不同类型的牛相关的基因，使用抑制性消减杂交（SSH）。通过对消减和未消减cDNA进行实时定量聚合酶链反应（qPCR）和对SSH PCR产物进行Southern印迹分析，对SSH的效率进行了评估，建立了12个SSH文库。我们确定了4个基因（CST3，GNAS，GPx4和PFN2）与牛的变化。使用来自SSH文库的70个表达的序列标签，用qPCR和Western印迹鉴定这些基因。克隆和测序使我们能够证明这4个基因与小鼠基因密切相关。我们可以假设这4个基因在牛的变异发展中起着重要作用。本研究为进一步研究牛脑W变异发生的相关基因及脑血管畸形的发生机制奠定了基础。
Deformities in the Circle of Willis (CoW) can significantly increase the risk of cerebrovascular disease in humans. However, the molecular mechanisms underlying these deformities have not been understood. Based on our previous studies, variations in the CoW of gerbils are hereditary. A normal CoW is observed in approximately 60% of gerbils, a percentage that also applies to humans. Thus, gerbil is an ideal experimental model for studying variations in the CoW. To study the mechanisms underlying these variations, we selected genes associated with different types of the CoW using suppression subtractive hybridization (SSH). After evaluating the efficiency of SSH using quantitative real-time polymerase chain reaction (qPCR) on subtracted and unsubtracted cDNA and Southern blotting on SSH PCR products, 12 SSH libraries were established. We identified 4 genes (CST3, GNAS, GPx4 and PFN2) associated with variations in the CoW. These genes were identified with qPCR and Western blotting using 70 expressed sequence tags from the SSH libraries. Cloning and sequencing allowed us to demonstrate that the 4 genes were closely related to mouse genes. We may assume that these 4 genes play an important role in the development of variations in the CoW. This study provides a foundation for further research of genes related to development of variations in the CoW and the mechanisms of dysmorphosis of cerebral vessels.
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影响因子：
3.4
作者：
Dreilich, M;Wagenius, G;Bergqvist, M
通讯作者：
Bergqvist, M
影响因子：
6.2
作者：
Portal, Orelvis;Izquierdo, Yovanny;Hofte, Monica
通讯作者：
Hofte, Monica
影响因子：
3.7
作者：
Hayward DC;Hetherington S;Behm CA;Grasso LC;Forêt S;Miller DJ;Ball EE
通讯作者：
Ball EE
影响因子：
4.1
作者：
Gardmo, C;Swerdlow, H;Mode, A
通讯作者：
Mode, A
影响因子：
50.3
作者：
Mouneimne G;Hansen SD;Selfors LM;Petrak L;Hickey MM;Gallegos LL;Simpson KJ;Lim J;Gertler FB;Hartwig JH;Mullins RD;Brugge JS
通讯作者：
Brugge JS