Mateos-Hernández Lourdes, Pipová Natália, Allain Eléonore, Henry Céline, Rouxel Clotilde, Lagrée Anne-Claire, Haddad Nadia, Boulouis Henri-Jean, Valdés James J, Alberdi Pilar, de la Fuente José, Cabezas-Cruz Alejandro, Šimo Ladislav
UMR BIPAR, Laboratoire de Santé Animale, ANSES, INRAE, Ecole Nationale Vétérinaire d'Alfort, Paris-Est Sup, 94700 Maisons-Alfort, France.
Faculty of Science, Pavol Jozef Šafarik University in Košice, 04180 Košice, Slovakia.
Pathogens. 2021 Jan 14;10(1):70. doi: 10.3390/pathogens10010070.
Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like embryonic cell line, ISE6, is an effective tool frequently used for examining tick-pathogen interactions. We detected 37 neuropeptide transcripts in the ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, . The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in -infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick-pathogen interactions.
神经肽是在蜱虫中枢神经系统即神经节中表达的小信号分子。神经元样胚胎细胞系ISE6是一种常用于研究蜱虫与病原体相互作用的有效工具。我们使用计算机方法在ISE6细胞系中检测到37种神经肽转录本，其中6种神经肽基因用于实验验证。在这6种神经肽基因中，ISE6细胞的速激肽相关肽（TRP）的转录本表达因蜱传病原体的感染菌株而异。TRP的免疫细胞化学显示在一个突出的ISE6细胞亚群中有细胞质表达。在感染的ISE6细胞中也证实了TRP的存在。神经节TRP的原位杂交和免疫组织化学显示在不同的神经元细胞中有表达。此外，在通过外周神经离开神经节的轴突以及与血神经相关的外侧节段器官中检测到TRP免疫反应。ISE6细胞中完整神经肽组的表征可能作为一种有效的体外工具，用于研究蜱传病原体如何与对蜱虫生理至关重要的神经节成分相互作用。因此，我们目前的研究是进一步研究蜱虫与病原体相互作用的神经元基础的体内实验的潜在垫脚石。
