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寄主植物和免疫刺激对毛虫唾液腺中化学感受蛋白和气味结合蛋白水平的影响。

Effect of host plant and immune challenge on the levels of chemosensory and odorant-binding proteins in caterpillar salivary glands.

作者信息

Celorio-Mancera Maria de la Paz, Ytterberg A Jimmy, Rutishauser Dorothea, Janz Niklas, Zubarev Roman A

机构信息

Stockholm University, Department of Zoology Ecology, Svante Arrheniusväg 18 B, 106 91, Stockholm, Sweden.

Karolinska Institute, Department of Medicine, Solna, Stockholm, Sweden.

出版信息

Insect Biochem Mol Biol. 2015 Jun;61:34-45. doi: 10.1016/j.ibmb.2015.04.006. Epub 2015 Apr 29.

Abstract

More than half of the proteome from mandibular glands in caterpillars is represented by chemosensory proteins. Based on sequence similarity, these proteins are putative transporters of ligands to gustatory receptors in sensory organs of insects. We sought to determine whether these proteins are inducible by comparing, both qualitatively and quantitatively, the salivary (mandibular and labial) proteomes from caterpillars (Vanessa cardui) reared on different plants and artificial diet containing either bacteria or bacterial cell-walls. We included a treatment where the caterpillars were switched from feeding on artificial diet to plant material at some point in their development. Additionally, we evaluated the degree of overlap between the proteomes in the hemolymph-filled coelom and salivary glands of caterpillars reared on plant material. We found that the quality and quantity of the identified proteins differed clearly between hemolymph-filled coelome, labial and mandibular glands. Our results indicated that even after molting and two-day feeding on a new diet, protein production is affected by the previous food source used by the caterpillar. Candidate proteins involved in chemosensory perception by insects were detected: three chemosensory (CSPs) and two odorant-binding proteins (OBPs). Using the relative amounts of these proteins across tissues and treatments as criteria for their classification, we detected hemolymph- and mandibular gland-specific CSPs and observed that their levels were affected by caterpillar diet. Moreover, we could compare the protein and transcript levels across tissues and treatment for at least one CSP and one OBP. Therefore, we have identified specific isoforms for testing the role of CSPs and OBPs in plant and pathogen recognition. We detected catalase, immune-related protein and serine proteases and their inhibitors in high relative levels in the mandibular glands in comparison to the labial glands. These findings suggest that the mandibular glands of caterpillars may play an important role protecting the caterpillar from oxidative stress, pathogens and aiding in digestion. Contamination with hemolymph proteins during dissection of salivary glands from caterpillars may occur but it is not substantial since the proteomes from hemolymph, mandibular and labial glands were easily discriminated from each other by principal component analysis of proteomic data.

摘要

毛虫下颌腺蛋白质组中超过一半由化学感受蛋白组成。基于序列相似性,这些蛋白质被认为是配体向昆虫感觉器官中味觉受体的转运体。我们试图通过定性和定量比较以不同植物以及含有细菌或细菌细胞壁的人工饲料饲养的毛虫(荨麻蛱蝶)的唾液(下颌腺和唇腺)蛋白质组,来确定这些蛋白质是否可被诱导。我们设置了一种处理方式,即让毛虫在发育的某个阶段从取食人工饲料转换为取食植物材料。此外,我们评估了取食植物材料的毛虫充满血淋巴的体腔和唾液腺中蛋白质组之间的重叠程度。我们发现,充满血淋巴的体腔、唇腺和下颌腺中鉴定出的蛋白质的质量和数量存在明显差异。我们的结果表明,即使在蜕皮并以新饲料喂养两天后,蛋白质的产生仍受毛虫先前食用的食物来源的影响。检测到了参与昆虫化学感受的候选蛋白质:三种化学感受蛋白(CSPs)和两种气味结合蛋白(OBPs)。以这些蛋白质在不同组织和处理中的相对含量作为分类标准,我们检测到了血淋巴和下颌腺特异性的CSPs,并观察到它们的水平受毛虫饮食的影响。此外,我们可以比较至少一种CSP和一种OBP在不同组织和处理中的蛋白质和转录水平。因此,我们已经鉴定出了用于测试CSPs和OBPs在植物和病原体识别中作用的特定亚型。与唇腺相比,我们在下颌腺中检测到过氧化氢酶、免疫相关蛋白和丝氨酸蛋白酶及其抑制剂的相对含量较高。这些发现表明,毛虫的下颌腺可能在保护毛虫免受氧化应激、病原体侵害以及帮助消化方面发挥重要作用。从毛虫唾液腺解剖过程中可能会发生血淋巴蛋白质污染,但由于通过蛋白质组学数据的主成分分析可以很容易地将血淋巴、下颌腺和唇腺的蛋白质组区分开来,所以这种污染并不严重。

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