College of Bioscience and Biotechnology, Shenyang Agricultural University, Liaoning Engineering and Technology Research Center for Insect Resource, Shenyang, People's Republic of China.
Bayer AG, Research & Development, Crop Science, Monheim am Rhein, Germany.
Arch Insect Biochem Physiol. 2024 Feb;115(2):e22089. doi: 10.1002/arch.22089.
Insecticide mode of action studies provide insights into how new insecticidal actives function and contribute to assessing safety to humans and nontarget organisms. Insect cell lines that express potential target sites can serve as valuable tools in this effort. In this paper, we report on the influence of two signaling molecules on protein expression in a nervous system cell line established from Spodoptera frugiperda (Bayer/BCIRL-SfNS2-0714-TR). We selected this line because we established it in our laboratory and we are experienced in using it. Cells were exposed to the insect developmental hormone (1 µg/mL 20-hydroxyecdysone, 20E) and/or a cyclooxygenase (COX) inhibitor (25 μM indomethacin, INDO; inhibits prostaglandin [PG] biosynthesis) for 24 h (Day 2), 72 h (Day 4), or 120 h (Day 6). We selected a PG biosynthesis inhibitor because PGs act in many aspects of insect biology, such as embryonic development, immunity, and protein phosphorylation. We selected the developmental hormone, 20E, because it also acts in fundamental aspects of insect biology. We identified specific proteins via in silico analysis. Changes in protein expression levels were determined using liquid chromatography-mass spectrometry (MS) + MS-MS. The largest number of changes in protein expression occurred on Day 2. The combination of 20E plus INDO led to 222 differentially expressed proteins, which documents the deep significance of PGs and 20E in insect biology. 20E and, separately, INDO led to changes in 30 proteins each (p value < 0.01; >2X or <0.5X-fold changes). We recorded changes in the expression of 9 or 12 proteins (20E), 10 or 6 proteins (INDO), and 21 or 20 proteins (20E + INDO) on D4 and D6, respectively. While the cell line was established from neuronal tissue, the differentially expressed proteins act in a variety of fundamental cell processes. In this paper, we moved beyond a list of proteins by providing detailed, Gene Ontology term analyses and enrichment, which offers an in-depth understanding of the influence of these treatments on the SfNS2 cells. Because proteins are active components of cell physiology in their roles as enzymes, receptors, elements of signaling transduction pathways, and cellular structures, changes in their expression levels under the influence of signaling molecules provide insights into their function in insect cell physiology.
杀虫剂作用模式的研究提供了有关新杀虫剂如何发挥作用以及有助于评估对人类和非目标生物安全性的见解。表达潜在靶标的昆虫细胞系可以作为这项工作的有价值的工具。在本文中，我们报告了两种信号分子对源自 Spodoptera frugiperda（Bayer/BCIRL-SfNS2-0714-TR）的神经系统细胞系中蛋白质表达的影响。我们选择这条线是因为它是我们实验室建立的，并且我们在使用它方面经验丰富。细胞暴露于昆虫发育激素（1μg/mL 20-羟基蜕皮酮，20E）和/或环氧化酶（COX）抑制剂（25μM 吲哚美辛，INDO；抑制前列腺素[PG]生物合成）24 小时（第 2 天）、72 小时（第 4 天）或 120 小时（第 6 天）。我们选择 PG 生物合成抑制剂是因为 PG 在昆虫生物学的许多方面发挥作用，例如胚胎发育、免疫和蛋白质磷酸化。我们选择发育激素 20E，是因为它也在昆虫生物学的基本方面发挥作用。我们通过计算机分析鉴定了特定蛋白质。使用液相色谱-质谱（MS）+MS-MS 测定蛋白质表达水平的变化。蛋白质表达变化最大的发生在第 2 天。20E 加 INDO 的组合导致 222 种差异表达的蛋白质，这证明了 PG 和 20E 在昆虫生物学中的深远意义。20E 和单独的 INDO 分别导致 30 种蛋白质的变化（p 值<0.01；2X 或 0.5X 倍变化）。我们记录了 D4 和 D6 时 9 或 12 种蛋白质（20E）、10 或 6 种蛋白质（INDO）和 21 或 20 种蛋白质（20E+INDO）的表达变化。虽然该细胞系是从神经元组织中建立的，但差异表达的蛋白质在各种基本的细胞过程中发挥作用。在本文中，我们通过提供详细的、基因本体论术语分析和富集，超越了蛋白质列表，深入了解了这些处理对 SfNS2 细胞的影响。由于蛋白质在作为酶、受体、信号转导途径元素和细胞结构的细胞生理活性成分中，其表达水平的变化为它们在昆虫细胞生理中的功能提供了见解。
