Increased expression of integrin and receptor tyrosine kinase genes during autograft fusion in the sponge Geodia cydonium.

W Wimmer, B Blumbach, B Diehl-Seifert, C Koziol, R Batel, R Steffen, I M Müller, W E Müller
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引用次数: 41

Abstract

Recently cDNAs coding for cell surface molecules have been isolated from sponges. The molecules for alpha-integrin, galectin, and receptor tyrosine kinase (RTK), obtained from the marine sponge, Geodia cydonium, have been described earlier. In the present study also the cDNA for one putative beta-integrin has been identified from G. cydonium. The deduced aa sequence comprises the characteristic signatures, found in other metazoan beta-integrin molecules; the estimated size is 95,215 Da. To obtain first insights into the molecular events which proceed during autograft fusion, the expressions of these genes were determined on transcriptional and translational level. The cDNAs as well as antibodies raised against the recombinant sponge proteins alpha-integrin, RTK and galectin were used and Northern blot experiments and immunocytochemical analyses have been performed. The results show that transcription of the two subunits of an integrin receptor as well as of the RTK are strongly upregulated after grafting; levels of > 10-fold have been determined in the fusion zone of the grafts after a 10 days incubation. Immunofluorescence studies of sections through the fusion zone support these data. In contrast the transcription of the gene encoding galectin is drastically downregulated after grafting. In a parallel series of experiments the level of the heat-shock protein-70 was determined and it was found that it remained unchanged after grafting. We conclude that integrin subunits and the RTK molecule are involved in self-self recognition of sponge.

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整合素和受体酪氨酸激酶基因在海地草自体移植物融合过程中的表达增加。
最近从海绵中分离出了编码细胞表面分子的cdna。α -整合素,半乳糖凝集素和受体酪氨酸激酶(RTK)的分子,从海绵中获得,Geodia cydonium,已经在前面描述过。在本研究中,还鉴定了一种推测的-整合素的cDNA。推导出的aa序列包含在其他后生动物β -整合素分子中发现的特征特征;估计大小为95,215 Da。为了获得在自体移植物融合过程中进行的分子事件的初步见解,在转录和翻译水平上确定了这些基因的表达。利用重组海绵蛋白α -整合素、RTK和凝集素的cdna和抗体,进行Northern blot实验和免疫细胞化学分析。结果表明,接枝后,整合素受体的两个亚基以及RTK的转录均显著上调;经过10天的培养,在移植物融合区确定了> 10倍的水平。通过融合区切片的免疫荧光研究支持这些数据。相反,编码凝乳素的基因的转录在嫁接后急剧下调。在平行的一系列实验中,测定了热休克蛋白70的水平,发现它在嫁接后保持不变。我们认为整合素亚基和RTK分子参与了海绵的自我识别。
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