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Serotonin signals through postsynaptic Gαq, Trio RhoGEF, and DAG to promote C. elegans egg-laying circuit activity and behavior

View ORCID ProfilePravat Dhakal, Sana Chaudhry, View ORCID ProfileKevin M. Collins
doi: https://doi.org/10.1101/2021.05.08.443256
Pravat Dhakal
Department of Biology, University of Miami, Coral Gables, FL, United States of America
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Sana Chaudhry
Department of Biology, University of Miami, Coral Gables, FL, United States of America
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Kevin M. Collins
Department of Biology, University of Miami, Coral Gables, FL, United States of America
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  • For correspondence: kevin.collins@miami.edu
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Abstract

Activated Gαq signals through Phospholipase-Cβ (PLCβ) and Trio, a Rho GTPase exchange factor (RhoGEF), but how these distinct effector pathways promote cellular responses to neurotransmitters like serotonin remains poorly understood. We used the egg-laying behavior circuit of C. elegans to determine whether PLCβ and Trio mediate serotonin and Gαq signaling through independent or related biochemical pathways. Using genetic rescue experiments, we find that PLCβ functions in neurons while Trio RhoGEF functions in both neurons and the postsynaptic vulval muscles. While Gαq, PLCβ, and Trio RhoGEF mutants all fail to lay eggs in response to serotonin, optogenetic stimulation of the serotonin releasing HSN command neurons restores egg laying only in PLCβ mutants. PLCβ mutants showed vulval muscle Ca2+ transients while strong Gαq and Trio RhoGEF mutants had little or no vulval muscle Ca2+ activity. Treatment with Phorbol esters that mimic Diacylglycerol (DAG), a product of PIP2 hydrolysis, rescued egg-laying circuit activity and behavior defects of Gαq signaling mutants, suggesting both Phospholipase C and Rho signaling promote synaptic transmission and egg-laying via DAG production. DAG activates effectors including UNC-13, however, we find that phorbol esters, but not serotonin, stimulate egg laying in unc-13 and PLCβ mutants. These results support a model where serotonin signaling through Gαq, PLCβ, and UNC-13 promote neurotransmitter release, and that serotonin also signals through Gαq, Trio RhoGEF, and an unidentified PMA-responsive effector to promote postsynaptic muscle excitability. Thus, the same neuromodulator serotonin can signal in distinct cells and effector pathways to coordinate activation of a motor behavior circuit.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted October 09, 2021.
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Serotonin signals through postsynaptic Gαq, Trio RhoGEF, and DAG to promote C. elegans egg-laying circuit activity and behavior
Pravat Dhakal, Sana Chaudhry, Kevin M. Collins
bioRxiv 2021.05.08.443256; doi: https://doi.org/10.1101/2021.05.08.443256
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Serotonin signals through postsynaptic Gαq, Trio RhoGEF, and DAG to promote C. elegans egg-laying circuit activity and behavior
Pravat Dhakal, Sana Chaudhry, Kevin M. Collins
bioRxiv 2021.05.08.443256; doi: https://doi.org/10.1101/2021.05.08.443256

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