DeLaney Kellen, Buchberger Amanda R, Atkinson Louise, Gründer Stefan, Mousley Angela, Li Lingjun
Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, WI 53706, USA.
School of Biological Sciences, Institute for Global Food Security, Queen's University Belfast, Belfast BT9 7BL, UK.
J Exp Biol. 2018 Feb 8;221(Pt 3):jeb151167. doi: 10.1242/jeb.151167.
Neuropeptides are one of the most diverse classes of signaling molecules and have attracted great interest over the years owing to their roles in regulation of a wide range of physiological processes. However, there are unique challenges associated with neuropeptide studies stemming from the highly variable molecular sizes of the peptides, low concentrations, high degree of structural diversity and large number of isoforms. As a result, much effort has been focused on developing new techniques for studying neuropeptides, as well as novel applications directed towards learning more about these endogenous peptides. The areas of importance for neuropeptide studies include structure, localization within tissues, interaction with their receptors, including ion channels, and physiological function. Here, we discuss these aspects and the associated techniques, focusing on technologies that have demonstrated potential in advancing the field in recent years. Most identification and structural information has been gained by mass spectrometry, either alone or with confirmations from other techniques, such as nuclear magnetic resonance spectroscopy and other spectroscopic tools. While mass spectrometry and bioinformatic tools have proven to be the most powerful for large-scale analyses, they still rely heavily on complementary methods for confirmation. Localization within tissues, for example, can be probed by mass spectrometry imaging, immunohistochemistry and radioimmunoassays. Functional information has been gained primarily from behavioral studies coupled with tissue-specific assays, electrophysiology, mass spectrometry and optogenetic tools. Concerning the receptors for neuropeptides, the discovery of ion channels that are directly gated by neuropeptides opens up the possibility of developing a new generation of tools for neuroscience, which could be used to monitor neuropeptide release or to specifically change the membrane potential of neurons. It is expected that future neuropeptide research will involve the integration of complementary bioanalytical technologies and functional assays.
神经肽是种类最多样化的信号分子类别之一，多年来因其在调节广泛生理过程中的作用而备受关注。然而，神经肽研究面临着独特的挑战，这些挑战源于肽分子大小高度可变、浓度低、结构多样性高以及异构体数量众多。因此，大量努力集中在开发研究神经肽的新技术以及旨在更多了解这些内源性肽的新应用上。神经肽研究的重要领域包括结构、在组织内的定位、与受体（包括离子通道）的相互作用以及生理功能。在这里，我们讨论这些方面及相关技术，重点关注近年来已显示出推动该领域发展潜力的技术。大多数鉴定和结构信息是通过质谱法单独获得的，或者是与其他技术（如核磁共振光谱法和其他光谱工具）的确认相结合获得的。虽然质谱法和生物信息学工具已被证明在大规模分析中最强大，但它们仍严重依赖互补方法进行确认。例如，组织内的定位可以通过质谱成像、免疫组织化学和放射免疫测定来探测。功能信息主要来自行为研究以及组织特异性测定、电生理学、质谱法和光遗传学工具。关于神经肽的受体，由神经肽直接门控的离子通道的发现为开发新一代神经科学工具开辟了可能性，这些工具可用于监测神经肽释放或特异性改变神经元的膜电位。预计未来的神经肽研究将涉及互补生物分析技术和功能测定的整合。
