Pathogen-associated molecular patterns activate expression of genes involved in cell proliferation, immunity and detoxification in the amebocyte-producing organ of the snail Biomphalaria glabrata.
Pathogen-associated molecular patterns activate expression of genes involved in cell proliferation, immunity and detoxification in the amebocyte-producing organ of the snail Biomphalaria glabrata.
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DOI：
10.1016/j.dci.2015.11.008
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发表时间：
2016-03
影响因子：
2.9
通讯作者：
Sullivan JT
中科院分区：
文献类型：
--
作者：
Zhang SM;Loker ES;Sullivan JT
光滑双脐螺的前心包壁已被确定为血细胞生成部位，因此被命名为变形细胞生成器官（APO）。多项研究表明，包括病原体相关分子模式（PAMPs）在内的外源性非生物和生物物质能够刺激APO的有丝分裂活性和/或增大其体积，这意味着APO在先天免疫中发挥作用。然而，这些反应背后的分子机制尚未得到探究，部分原因是难以获取足够的APO组织用于基因表达研究。通过使用改良的RNA提取技术和微阵列技术，我们研究了在注射两种细菌PAMPs（脂多糖（LPS）和肽聚糖（PGN））或岩藻依聚糖（FCN）24小时后，从蜗牛体内解剖出的APO的转录组反应。FCN可能模拟了曼氏血吸虫胞蚴上富含岩藻糖基的聚糖PAMPs。根据差异表达基因的数量，与注射生理盐水的对照组相比，LPS表现出最强的活性。我们观察到参与细胞增殖、免疫反应和解毒代谢的基因同时被激活。一种编码细胞周期检查点1激酶（有丝分裂的关键调节因子）的基因在LPS刺激后高度表达。此外，还发现七种在蛋白质合成中起关键作用的不同氨酰-tRNA合成酶高度表达。除了刺激参与细胞增殖的基因外，注射的物质，尤其是LPS，还诱导了许多免疫相关基因的表达，包括精氨酸酶、短肽聚糖识别蛋白、肿瘤坏死因子受体、纤维胶凝蛋白、钙调蛋白、细菌通透增强蛋白和E3泛素蛋白连接酶。重要的是，我们观察到四个GiMAP（免疫相关蛋白GTP酶）基因显著上调，这一结果首次为GiMAP在原口动物中的免疫作用提供了证据。此外，注射物质还引发了编码细胞色素P450、谷胱甘肽-S-转移酶、多药耐药蛋白的基因以及大量编码与降解和解毒代谢相关酶的基因表达发生改变 。
The anterior pericardial wall of the snail Biomphalaria glabrata has been identified as a site of hemocyte production, hence has been named the amebocyte-producing organ (APO). A number of studies have shown that exogenous abiotic and biotic substances, including pathogen associated molecular patterns (PAMPs), are able to stimulate APO mitotic activity and/or enlarge its size, implying a role for the APO in innate immunity. The molecular mechanisms underlying such responses have not yet been explored, in part due to the difficulty in obtaining sufficient APO tissue for gene expression studies. By using a modified RNA extraction technique and microarray technology, we investigated transcriptomic responses of APOs dissected from snails at 24 hours post-injection with two bacterial PAMPs, lipopolysaccharide (LPS) and peptidoglycan (PGN), or with fucoidan (FCN), which may mimic fucosyl-rich glycan PAMPs on sporocysts of Schistosoma mansoni. Based upon the number of genes differentially expressed, LPS exhibited the strongest activity, relative to saline-injected controls. A concurrent activation of genes involved in cell proliferation, immune response and detoxification metabolism was observed. A gene encoding checkpoint 1 kinase, a key regulator of mitosis, was highly expressed after stimulation by LPS. Also, seven different aminoacyl-tRNA synthetases that play an essential role in protein synthesis were found to be highly expressed. In addition to stimulating genes involved in cell proliferation, the injected substances, especially LPS, also induced expression of a number of immune-related genes including arginase, peptidoglycan recognition protein short form, tumor necrosis factor receptor, ficolin, calmodulin, bacterial permeability increasing proteins and E3 ubiquitin-protein ligase. Importantly, significant up-regulation was observed in four GiMAP (GTPase of immunity-associated protein) genes, a result which provides the first evidence suggesting an immune role of GiMAP in protostome animals. Moreover, altered expression of genes encoding cytochrome P450, glutathione-S-transferase, multiple drug resistance protein as well as a large number of genes encoding enzymes associated with degradation and detoxification metabolism was elicited in response to the injected substances.