Indomethacin and 20‐hydroxyecdysone influence protein expression in a Spodoptera frugiperda nervous system cell line

Author:

Wang Yong1ORCID,Buer Benjamin2ORCID,Goodman Cynthia L.3ORCID,Kang David3ORCID,Reall Tamra4,Dohn Susanne2ORCID,Ringbauer Joseph3ORCID,Li Yaofa5ORCID,Geibel Sven2,Stanley David3ORCID

Affiliation:

1. College of Bioscience and Biotechnology Shenyang Agricultural University, Liaoning Engineering and Technology Research Center for Insect Resource Shenyang People's Republic of China

2. Bayer AG, Research & Development, Crop Science Monheim am Rhein Germany

3. Biological Control of Insects Research Laboratory USDA/Agricultural Research Service Columbia Missouri USA

4. University of Missouri‐Extension Blue Springs Missouri USA

5. Plant Protection Institute, Hebei Academy of Agricultural and Forestry Sciences/IPM Center of Hebei Province/Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China Ministry of Agriculture Baoding China

Abstract

AbstractInsecticide 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.

Funder

U.S. Department of Agriculture

Agricultural Research Service

Publisher

Wiley

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