Neuropeptides can have significant effects on neurons and synapses, but among the ∼250 predicted peptides in nematodes, few have been characterized functionally. Here, we report new neuropeptides in the 4 RME nerve ring motorneurons of the nematode Ascaris suum. These GABAergic neurons are involved in three-dimensional head movement. Mass spectrometry (MS) of single dissected RMEs detected a total of 12 neuropeptides (encoded by five genes), nine of which are novel. None of these are expressed in the DI/VI inhibitory GABAergic motorneurons that synapse onto body wall muscle. Using peptide sequences obtained by tandem MS, we cloned the peptide-encoding transcripts and synthesized riboprobes for in situ hybridization (ISH). This complementary technique corroborated the results from single-cell MS, showing that the dissections were not contaminated with adhering tissue from other cells. We also synthesized a multiple antigenic peptide to raise a highly specific antibody against one of the endogenous peptides, which labeled the same cells detected by MS and ISH. Our results show that the RMEs can be divided into two subsets: RMED/V (expressing afp-2, afp-15, Asu-nlp-58, and high levels of afp-16) and RMEL/R (expressing afp-15 and low levels of afp-4 and afp-16). Almost all of these peptides are bioactive in A. suum.

中文翻译：

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不同的生物活性神经肽表达的GABA能神经RME环运动神经元的两个子类中的蛔虫

神经肽可能对神经元和突触有重大影响，但在线虫中预测的约250种肽中，很少有功能性的表征。在这里，我们报告中的线虫的4 RME神经环运动神经元的新神经肽猪蛔虫。这些GABA能神经元参与三维头部运动。单个解剖的RME的质谱（MS）检测到总共12种神经肽（由5个基因编码），其中9种是新颖的。这些都不在突触到体壁肌肉上的DI / VI抑制性GABA能动神经元中表达。使用通过串联质谱获得的肽序列，我们克隆了编码肽的转录本，并合成了核糖核酸以进行原位杂交（ISH）。这项补充技术证实了单细胞MS的结果，表明该解剖未受到其他细胞粘附组织的污染。我们还合成了一种多抗原肽，以产生针对一种内源肽的高度特异性抗体，该抗体标记了通过MS和ISH检测到的相同细胞。afp-2，afp-15，Asu-nlp-58和高水平的afp-16）和RMEL / R（表示afp-15和低水平的afp-4和afp-16）。这些肽中几乎所有的肽在su.suum中都具有生物活性。