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Published December 2017 | Supplemental Material
Journal Article Open

Comparative analysis of gene expression profiles for several migrating cell types identifies cell migration regulators

Abstract

Cell migration is an instrumental process that ensures cells are properly positioned to support the specification of distinct tissue types during development. To provide insight, we used fluorescence activated cell sorting (FACS) to isolate two migrating cell types from the Drosophila embryo: caudal visceral mesoderm (CVM) cells, precursors of longitudinal muscles of the gut, and hemocytes (HCs), the Drosophila equivalent of blood cells. ~ 350 genes were identified from each of the sorted samples using RNA-seq, and in situ hybridization was used to confirm expression within each cell type or, alternatively, within other interacting, co-sorted cell types. To start, the two gene expression profiling datasets were compared to identify cell migration regulators that are potentially generally-acting. 73 genes were present in both CVM cell and HC gene expression profiles, including the transcription factor zinc finger homeodomain-1 (zfh1). Comparisons with gene expression profiles of Drosophila border cells that migrate during oogenesis had a more limited overlap, with only the genes neyo (neo) and singed (sn) found to be expressed in border cells as well as CVM cells and HCs, respectively. Neo encodes a protein with Zona pellucida domain linked to cell polarity, while sn encodes an actin binding protein. Tissue specific RNAi expression coupled with live in vivo imaging was used to confirm cell-autonomous roles for zfh1 and neo in supporting CVM cell migration, whereas previous studies had demonstrated a role for Sn in supporting HC migration. In addition, comparisons were made to migrating cells from vertebrates. Seven genes were found expressed by chick neural crest cells, CVM cells, and HCs including extracellular matrix (ECM) proteins and proteases. In summary, we show that genes shared in common between CVM cells, HCs, and other migrating cell types can help identify regulators of cell migration. Our analyses show that neo in addition to zfh1 and sn studied previously impact cell migration. This study also suggests that modification of the extracellular milieu may be a fundamental requirement for cells that undergo cell streaming migratory behaviors.

Additional Information

© 2017 Elsevier B.V. Received date: 7 January 2017. Revised date: 13 April 2017. Accepted date: 13 April 2017. We thank Katja Brückner, Manfred Frasch, and Jennifer Lippincott-Schwartz for sharing Drosophila stocks and probes. We are grateful to Igor Antoshechkin in the Millard and Muriel Jacobs Genetics and Genomics Laboratory at California Institute of Technology for sequencing support and answering questions on data analysis. We also thank Rochelle Diamond at the Caltech Flow Cytometry Facility for excellent assistance in cell sorting. We additionally appreciate Justin Bois for his data analysis advice. This work was supported by National Institute of Health grants R01GM1043838 to A.S., 1F32GM119395-01 to F.M., and 5T32GM007616-38 to H.C., and by American Heart Association postdoctoral fellowship grant 11POST7600181 to Y.-K.B. Author contributions: Y.-K.B. and A.S. devised the overall question and experimental approach; Y.-K.B conducted the FACS experiments and data analysis; F.M., H·C., and Y.-K.B. performed in situ hybridization and RNAi experiments; Y.-K.B., F.M, H.C., and A.S. analyzed the data; and Y.-K.B., F.M, H.C, and A.S. wrote the paper.

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Supplemental Material - 1-s2.0-S0925477316301265-mmc1.docx

Supplemental Material - 1-s2.0-S0925477316301265-mmc2.mp4

Supplemental Material - 1-s2.0-S0925477316301265-mmc3.mp4

Supplemental Material - 1-s2.0-S0925477316301265-mmc4.mp4

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Created:
August 21, 2023
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