Extrasynaptic acetylcholine signaling through a muscarinic receptor regulates cell migration
Abstract
Acetylcholine (ACh) promotes various cell migrations in vitro, but there are few investigations into this nonsynaptic role of ACh signaling in vivo. Here we investigate the function of a muscarinic receptor on an epithelial cell migration in Caenorhabditis elegans. We show that the migratory gonad leader cell, the linker cell (LC), uses an M1/M3/M5-like muscarinic ACh receptor GAR-3 to receive extrasynaptic ACh signaling from cholinergic neurons for its migration. Either the loss of the GAR-3 receptor in the LC or the inhibition of ACh release from cholinergic neurons resulted in migratory path defects. The overactivation of the GAR-3 muscarinic receptor caused the LC to reverse its orientation through its downstream effectors Gαq/egl-30, PLCβ/egl-8, and TRIO/unc-73. This reversal response only occurred in the fourth larval stage, which corresponds to the developmental time when the GAR-3::yellow fluorescent protein receptor in the membrane relocalizes from a uniform to an asymmetric distribution. These findings suggest a role for the GAR-3 muscarinic receptor in determining the direction of LC migration.
Additional Information
© 2021 The Author(s). Published under the PNAS license. Contributed by Paul W. Sternberg, September 6, 2020 (sent for review March 20, 2019; reviewed by Reinhard W. Köster and James B. Rand) Published in issue January 5, 2021. Published first December 23, 2020. We thank laboratory members, particularly Pei Yin Shih, Ravi Nath, and Han Wang, for comments on the paper. Some strains were provided by the Caenorhabditis Genetics Center, which is funded by NIH Office of Research Infrastructure Programs (grant P40 OD010440), and by the C. elegans Gene Knockout Project at the Oklahoma Medical Research Foundation, which was part of the International C. elegans Gene Knockout Consortium. For image collection and analysis, we used the Caltech Biological Imaging Facility and the Center for Advanced Methods in Biological Image Analysis. We received microscopy training from Dr. Andres Collazo and Huygens software training from Say-Tar Goh. Dr. Rene Garcia, Dr. Derek Sieburth, and Dr. Kenneth Miller kindly provided strains and reagents. This work was supported by HHMI with whom P.W.S. was an investigator and by US Public Health Service NIH, Eunice Kennedy Shriver Institute of Child Health and Human Development grant R01 HD091327 to P.W.S. and M.K. Data Availability. All study data are included in the article and supporting information. Author contributions: M.K., I.K., and P.W.S. designed research; M.K., I.K., and S.G. performed research; M.K. and A.C. contributed new reagents/analytic tools; M.K., I.K., and A.C. analyzed data; and M.K. and P.W.S. wrote the paper. Reviewers: R.W.K., Technische Universität Braunschweig; and J.B.R., University of Oklahoma Health Sciences Center. The authors declare no competing interest. This article contains supporting information online at https://www.pnas.org/lookup/suppl/doi:10.1073/pnas.1904338118/-/DCSupplemental.Attached Files
Published - e1904338118.full.pdf
Supplemental Material - pnas.1904338118.sapp.pdf
Supplemental Material - pnas.1904338118.sd01.xlsx
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Additional details
- PMCID
- PMC7817160
- Eprint ID
- 107278
- Resolver ID
- CaltechAUTHORS:20201224-085807779
- P40 OD010440
- NIH
- Oklahoma Medical Research Foundation
- 047-101
- Howard Hughes Medical Institute (HHMI)
- R01HD091327
- NIH
- Created
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2021-01-04Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering