Iovinella Immacolata, Ban Liping, Song Limei, Pelosi Paolo, Dani Francesca Romana
Biology Department, University of Firenze, via Madonna del Piano 6, 50019 Sesto Fiorentino, Italy.
Department of Grassland Science, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
Insect Biochem Mol Biol. 2016 Nov;78:58-68. doi: 10.1016/j.ibmb.2016.09.004. Epub 2016 Sep 29.
In arthropods, the large majority of studies on olfaction have been focused on insects, where most of the proteins involved have been identified. In particular, chemosensing in insects relies on two families of membrane receptors, olfactory/gustatory receptors (ORs/GRs) and ionotropic receptors (IRs), and two classes of soluble proteins, odorant-binding proteins (OBPs) and chemosensory proteins (CSPs). In other arthropods, such as ticks and mites, only IRs have been identified, while genes encoding for OBPs and CSPs are absent. A third class of soluble proteins, called Niemann-Pick C2 (NPC2) has been suggested as potential carrier for semiochemicals both in insects and other arthropods. Here we report the results of a proteomic analysis on olfactory organs (Haller's organ and palps) and control tissues of the tick Ixodes ricinus, and of immunostaining experiments targeting NPC2s. Adopting different extraction and proteomic approaches, we identified a large number of proteins, and highlighted those differentially expressed. None of the 13 NPC2s known for this species was found. On the other hand, using immunocytochemistry, we detected reaction against one NPC2 in the Haller's organ and palp sensilla. We hypothesized that the low concentration of such proteins in the tick's tissues could possibly explain the discrepant results. In ligand-binding assays the corresponding recombinant NPC2 showed good affinity to the fluorescent probe N-phenylnaphthylamine and to few organic compounds, supporting a putative role of NPC2s as odorant carriers.
在节肢动物中,绝大多数关于嗅觉的研究都集中在昆虫上,其中涉及的大多数蛋白质已被鉴定出来。特别是,昆虫的化学感应依赖于两类膜受体,即嗅觉/味觉受体(ORs/GRs)和离子型受体(IRs),以及两类可溶性蛋白质,即气味结合蛋白(OBPs)和化学感应蛋白(CSPs)。在其他节肢动物中,如蜱虫和螨虫,仅鉴定出了IRs,而编码OBPs和CSPs的基因则不存在。有人提出,第三类可溶性蛋白质,即尼曼-皮克C2型蛋白(NPC2),在昆虫和其他节肢动物中可能是信息素的潜在载体。在此,我们报告了对蓖麻蜱嗅觉器官(哈勒氏器和触须)以及对照组织进行蛋白质组学分析的结果,以及针对NPC2s的免疫染色实验结果。采用不同的提取和蛋白质组学方法,我们鉴定出了大量蛋白质,并突出了那些差异表达的蛋白质。在该物种已知的13种NPC2s中均未发现。另一方面,通过免疫细胞化学方法,我们在哈勒氏器和触须感器中检测到了针对一种NPC2的反应。我们推测,蜱虫组织中此类蛋白质的低浓度可能解释了结果的差异。在配体结合试验中,相应的重组NPC2对荧光探针N-苯基萘胺和少数有机化合物表现出良好的亲和力,这支持了NPC2s作为气味载体的假定作用。