Transcriptomic profiling of sex-specific olfactory neurons reveals subset-specific receptor expression in C. elegans
Abstract
The nematode Caenorhabditis elegans utilizes chemosensation to navigate an ever-changing environment for its survival. A class of secreted small-molecule pheromones, termed ascarosides, play an important role in olfactory perception by affecting biological functions ranging from development to behavior. The ascaroside ascr#8 mediates sex-specific behaviors, driving avoidance in hermaphrodites and attraction in males. Males sense ascr#8 via the ciliated male-specific cephalic sensory (CEM) neurons, which exhibit radial symmetry along dorsal-ventral and left-right axes. Calcium imaging studies suggest a complex neural coding mechanism that translates stochastic physiological responses in these neurons to reliable behavioral outputs. To test the hypothesis that neurophysiological complexity arises from differential expression of genes, we performed cell-specific transcriptomic profiling; this revealed between 18 and 62 genes with at least two-fold higher expression in a specific CEM neuron type versus both other CEM neurons and adult males. These included two G protein coupled receptor (GPCR) genes, srw-97 and dmsr-12, that were specifically expressed in non-overlapping subsets of CEM neurons and whose expression was confirmed by GFP reporter analysis. Single CRISPR-Cas9 knockouts of either srw-97 or dmsr-12 resulted in partial defects, while a double knockout of both srw-97 and dmsr-12 completely abolished the attractive response to ascr#8. Together, our results suggest that the evolutionarily distinct GPCRs SRW-97 and DMSR-12 act non-redundantly in discrete olfactory neurons to facilitate male-specific sensation of ascr#8.
Additional Information
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. We thank the Caenorhabditis Genetics Center, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD01044), as well as the National BioResource Project, Ding Xue (University of Colorado, Boulder), L. Rene Garcia (Texas A&M University), and Douglas Portman (University of Rochester Medical Center) for providing strains. We also thank Igor Antoshechkin, Caltech Genomics facility for sequencing and InVivo Biosystems for generating transgenic and CRISPR knockout animals. The synthetic ascr#8 utilized in this study was provided by Frank Schroeder (Cornell University). The research reported in this publication was supported by NIH R01 DC016058 (J.S.), R01 GM084389 (P.W.S.), the Howard Hughes Medical Institute (P.W.S.), Moore Foundation Grant No. 4551 (E.M.S.), and Cornell startup funding (E.M.S.). We thank Titus Brown and the Michigan State University High-Performance Computing Center (supported by U.S. Department of Agriculture grant 2010-65205-20361 and NIFA–National Science Foundation (NSF) grant IOS-0923812) for computational support; additional computing was enabled by start-up and research allocations from NSF XSEDE (TG538 MCB180039 and TG-MCB190010). Data availability: RNA-seq reads have been deposited in the NCBI Sequence Read Archive (SRA) under the BioProject accession number PRJNA781271. The authors have declared no competing interest.Attached Files
Submitted - 2021.10.26.465928v2.full.pdf
Supplemental Material - media-1.pdf
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Additional details
- Alternative title
- Male pheromone G protein-coupled receptors in C. elegans
- Eprint ID
- 111706
- DOI
- 10.1101/2021.10.26.465928
- Resolver ID
- CaltechAUTHORS:20211102-173632613
- NIH
- P40 OD010440
- NIH
- R01 DC016058
- NIH
- R01 GM084389
- Howard Hughes Medical Institute (HHMI)
- Gordon and Betty Moore Foundation
- 4551
- Cornell University
- Department of Agriculture
- 2010-65205-20361
- NSF
- IOS-0923812
- NSF
- TG538 MCB180039
- NSF
- TG-MCB190010
- Created
-
2021-11-02Created from EPrint's datestamp field
- Updated
-
2023-03-17Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering (BBE)