Abstract
Using the 10x Genomics Chromium single-cell RNA sequencing (scRNA-seq) platform, we discovered unexpected heterogeneity in an established cell line developed from the midgut of the Fall armyworm, Spodoptera frugiperda, a major global pest. We analyzed the sequences of 18,794 cells and identified ten unique cellular clusters, including stem cells, enteroblasts, enterocytes and enteroendocrine cells, characterized by the expression of specific marker genes. Additionally, these studies addressed an important knowledge gap by investigating the expression of genes coding for respiratory and midgut membrane insecticide targets classified by the Insecticide Resistance Action Committee. Dual-fluorescence tagging method, fluorescence microscopy and fluorescence- activated cell sorting confirmed the expression of midgut cell type-specific genes. Stem cells were isolated from the heterogeneous population of SfMG-0617 cells. Our results, validated by KEGG and Gene Ontology analyses and supported by Monocle 3.0, advance the fields of midgut cellular biology and establish standards for scRNA-seq studies in non-model organisms.
Document Type
Article
Publication Date
9-2024
Digital Object Identifier (DOI)
https://doi.org/10.1016/j.ygeno.2024.110898
Funding Information
This material is based on the work supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technolo- gies (under grant no. IIP-1821936 and by industry partners), and the US Department of Agriculture (under HATCH Project 2353057000). We thank the College of Arts & Sciences Imaging Center and the Flow Cytometry and Immune Monitoring Core for technical assistance with scRNA-seq and flow cytometry analysis, respectively. The A&S Imaging Center is supported by the UK College of Arts & Sciences and the Department of Biology. The UK Flow Cytometry & Immune Monitoring core facility is partly supported by the Office of the Vice President for Research, the Markey Cancer Center, and an NCI Center Core Support Grant (P30 CA177558) to the University of Kentucky Markey Cancer Center.
Repository Citation
Arya, Surjeet Kumar; Harrison, Douglas A.; and Palli, Subba R., "Deciphering cellular heterogeneity in Spodoptera frugiperda midgut cell line through single cell RNA sequencing" (2024). Entomology Faculty Publications. 243.
https://uknowledge.uky.edu/entomology_facpub/243
Notes/Citation Information
0888-7543/© 2024 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/by- nc/4.0/).