Allograft rejection in the mixed cell reaction system of the demosponge Suberites domuncula is controlled by differential expression of apoptotic genes
Allograft rejection in the mixed cell reaction system of the demosponge Suberites domuncula is controlled by differential expression of apoptotic genes
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DOI：
10.1007/s00251-004-0718-6
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发表时间：
2004-11-01
期刊：
影响因子：
3.2
通讯作者：
Müller, WEG
中科院分区：
文献类型：
--
作者：
Wiens, M;Peroviç-Ottstadt, S;Müller, WEG
直到最近，分子探针的缺乏阻碍了海绵中促凋亡和抗凋亡基因表达的测定。在解决这个问题的方法中，本研究描述了多种cDNA从demosponge Suberites domuncula，编码的蛋白质是特征性的启动细胞凋亡（半胱天冬酶，MA 3，ALG-2蛋白），预防程序性细胞死亡（2 Bcl-2同源蛋白，FAIM相关多肽，DAD-1相关蛋白），和形态发生过程（类维生素A X受体）。将它们用作探针，在异基因混合海绵细胞反应（MSCR）系统中体外监测表达水平。在同种异体MSCR中，将来自相同物种的遗传上不同的动物的两细胞聚集体（原型体）彼此相邻放置。在培养约8天后，其中一个原态体经历了凋亡性死亡，而第二个仍然存活。通过北方印迹和原位杂交测定上述基因的表达水平。这些实验表明，在凋亡的原态，表达的特征性凋亡基因，而在非凋亡聚集体的细胞存活基因高度上调。有趣的是，在该测定中，类维生素A X受体的转录水平在凋亡原型体中比在非凋亡聚集体中高。我们的数据首次表明，在体外MSCR系统中，同种异体识别导致一个伴侣的凋亡细胞死亡，而另一个存活。我们认为，这一过程是由一个差异表达的促凋亡和促生存基因在这里研究。
Until recently, the lack of molecular probes hampered the determination of the expression of pro-apoptotic and anti-apoptotic genes in sponge. In an approach to solve this problem, the present study describes a variety of cDNAs from the demosponge Suberites domuncula, coding for proteins that are characteristic for the initiation of apoptosis (caspase, MA3, ALG-2 protein), for the prevention of programmed cells death (2 Bcl-2 homology proteins, FAIM-related polypeptide, and DAD-1-related protein), and for morphogenetic processes (retinoid X receptor). They were used as probes to monitor the expression levels in vitro in the allogeneic mixed sponge cell reaction (MSCR) system. In the allogeneic MSCR, two-cell aggregates (primmorphs) from genetically different animals of the same species were positioned next to each other. After approximately 8 days in culture, one of the primmorphs underwent apoptotic death, while the second remained alive. The expression levels of the aforementioned genes were determined by Northern blotting and by in situ hybridization. These experiments revealed that in the apoptotic primmorph, the characteristic apoptotic genes were expressed, while in the non-apoptotic aggregates the cell-survival genes are highly upregulated. Interestingly, the transcript levels of retinoid X receptor were higher in apoptotic primmorphs than in the non-apoptotic aggregate in the assay. Our data show for the first time that in the in vitro MSCR system, allogeneic recognition led to apoptotic cell death in one partner, while the other one survived. We suggest that this process is controlled by a differential expression of the pro-apoptotic and pro-survival genes studied here.