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蝗虫中气味结合蛋白和化学感应蛋白的表达与免疫定位

Expression and immunolocalisation of odorant-binding and chemosensory proteins in locusts.

作者信息

Jin X, Brandazza A, Navarrini A, Ban L, Zhang S, Steinbrecht R A, Zhang L, Pelosi P

机构信息

Department of Entomology, China Agricultural University, Beijing.

出版信息

Cell Mol Life Sci. 2005 May;62(10):1156-66. doi: 10.1007/s00018-005-5014-6.

DOI:10.1007/s00018-005-5014-6
PMID:15928808
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11924489/
Abstract

We have identified, cloned and expressed a new chemosensory protein (CSP) in the desert locust Schistocerca gregaria belonging to a third sub-class of these polypeptides. Polyclonal antibodies stained a band of 14 kDa, as expected, in the extracts of antennae and palps of the adults, but not in the 4th and 5th instars. In the related species Locusta migratoria, instead, the same antibodies cross-reacted only with a band of apparent molecular mass of 35 kDa in the extract of 1st-5th instars, but not in the adults. The recombinant protein binds the fluorescent probe N-phenyl-1-naphthylamine, but none of the compounds so far reported as pheromones for S. gregaria. The expression of the odorant-binding protein (OBP) and of CSPs of sub-classes I and II was also monitored in antennae, tarsi, palpi, wings and other organs of solitary and gregarious locusts in their nymphal and adult stages. OBP was found to be antenna specific, where it is expressed at least from the 3rd instar in both solitary and gregarious locusts. CSPs, instead, appear to be more ubiquitous, with different expression patterns, according to the sub-class. Immunocytochemistry experiments revealed that OBP is present in the sensillum lymph of sensilla trichodea and basiconica, while CSP-I and CSP-III were found in the outer sensillum lymph of sensilla chaetica and in the sub-cuticular space between epidermis and cuticle of the antenna. Sensilla chaetica on other parts of the body showed the same expression of CSP-I as those on the antenna.

摘要

我们已经在沙漠蝗(Schistocerca gregaria)中鉴定、克隆并表达了一种新的化学感受蛋白(CSP),它属于这些多肽的第三个亚类。如预期的那样,多克隆抗体在成年沙漠蝗触角和须肢的提取物中染出了一条14 kDa的条带,但在四龄和五龄若虫中未染出。相反,在相关物种飞蝗(Locusta migratoria)中,相同的抗体仅在一龄至五龄若虫的提取物中与一条表观分子量为35 kDa的条带发生交叉反应,而在成虫中未发生交叉反应。重组蛋白能结合荧光探针N-苯基-1-萘胺,但不能结合迄今为止报道的任何一种沙漠蝗信息素化合物。我们还监测了独居和群居沙漠蝗若虫及成虫阶段触角、跗节、须肢、翅膀和其他器官中气味结合蛋白(OBP)以及I类和II类CSP的表达情况。发现OBP具有触角特异性,在独居和群居沙漠蝗中至少从三龄开始表达。相反,CSP似乎分布更广泛,根据亚类不同具有不同的表达模式。免疫细胞化学实验表明,OBP存在于毛形感器和锥形感器的感器淋巴中,而CSP-I和CSP-III则存在于刺形感器的外部感器淋巴以及触角表皮与角质层之间的皮下间隙中。身体其他部位的刺形感器显示出与触角上的刺形感器相同的CSP-I表达。