Molecular characterization of frog vocal neurons using constellation pharmacology
Molecular characterization of frog vocal neurons using constellation pharmacology
复制标题
使用星座药理学对青蛙发声神经元进行分子表征
DOI：
10.1152/jn.00105.2020
复制
发表时间：
2020
影响因子：
2.5
通讯作者：
Yamaguchi, Ayako
中科院分区：
文献类型：
--
作者：
Inagaki, Ryota Thomas;Raghuraman, Shrinivasan;Chase, Kevin;Steele, Theresa;Zornik, Erik;Olivera, Baldomero M;Yamaguchi, Ayako
运动回路中神经元细胞类别的识别和表征对于理解行为的神经基础是必不可少的。鉴定功能大分子的细胞特异性表达是一项具有挑战性的任务，特别是在非遗传模式生物中。在这里，我们进行了星座药理学，钙离子成像的解离神经元，以确定功能受体表达的成年男性和女性非洲爪蛙，非洲爪蟾的发声神经元。以前，我们确定了一个群体的发声神经元称为快速颤音神经元（FTNs）在两栖动物臂旁核（PB），表达N-甲基-D-天冬氨酸（NMDA）受体和GABA和/或甘氨酸受体。使用星座药理学，我们确定了四种细胞类别的推定快速颤音神经元（pFTNs，响应NMDA和GABA/甘氨酸应用程序）。我们发现，一些pFTNs响应于P物质（SP），乙酰胆碱（ACh），或两者的应用。使用离体制剂从FTN获得的电生理记录证实SP和/或ACh使FTN脱髓鞘。将ACh、SP或它们的拮抗剂注入PBs表明，ACh受体对发声不是充分的，而是必需的，SP受体在形成发声形态中起作用。此外，我们还发现成年女性的PB。尽管它们的发声方式在性别上不同，但它们也包含了所有神经元的子类，频率与雄性相似。这些结果揭示了调节X的新型神经调质。laevisvocal生产和展示星座药理学的力量，在识别神经元亚型标记的细胞特异性受体的功能表达在非遗传模型organism.NEW &值得注意的神经元的分子概况是理解神经元功能的关键，但他们的识别是具有挑战性的，特别是在非遗传模型的有机体。在这里，我们的特点是在发声的非洲爪蛙，非洲爪蟾神经元膜大分子的功能表达，使用一种技术称为星座药理学。我们发现，乙酰胆碱和/或P物质受体表达的某些类别的发声神经元，它们的激活在正常发声的生产中发挥作用。
Identification and characterization of neuronal cell classes in motor circuits are essential for understanding the neural basis of behavior. It is a challenging task, especially in a non-genetic-model organism, to identify cell-specific expression of functional macromolecules. Here, we performed constellation pharmacology, calcium imaging of dissociated neurons to pharmacologically identify functional receptors expressed by vocal neurons in adult male and female African clawed frogs,Xenopus laevis. Previously we identified a population of vocal neurons called fast trill neurons (FTNs) in the amphibian parabrachial nucleus (PB) that expressN-methyl-d-aspartate (NMDA) receptors and GABA and/or glycine receptors. Using constellation pharmacology, we identified four cell classes of putative fast trill neurons (pFTNs, responsive to both NMDA and GABA/glycine applications). We discovered that some pFTNs responded to the application of substance P (SP), acetylcholine (ACh), or both. Electrophysiological recordings obtained from FTNs using an ex vivo preparation verified that SP and/or ACh depolarize FTNs. Bilateral injection of ACh, SP, or their antagonists into PBs showed that ACh receptors are not sufficient but necessary for vocal production, and SP receptors play a role in shaping the morphology of vocalizations. Additionally, we discovered that the PB of adult femaleX. laevisalso contains all the subclasses of neurons at a similar frequency as in males, despite their sexually distinct vocalizations. These results reveal novel neuromodulators that regulateX. laevisvocal production and demonstrate the power of constellation pharmacology in identifying the neuronal subtypes marked by functional expression of cell-specific receptors in non-genetic-model organisms.NEW & NOTEWORTHYMolecular profiles of neurons are critical for understanding the neuronal functions, but their identification is challenging especially in non-genetic-model organisms. Here, we characterized the functional expression of membrane macromolecules in vocal neurons of African clawed frogs,Xenopus laevis, using a technique called constellation pharmacology. We discovered that receptors for acetylcholine and/or substance P are expressed by some classes of vocal neurons, and their activation plays a role in the production of normal vocalizations.