4.8 Article

High-performance optical control of GPCR signaling by bistable animal opsins MosOpn3 and LamPP in a molecular propertydependent manner

出版社

NATL ACAD SCIENCES
DOI: 10.1073/pnas.2204341119

关键词

rhodopsin; second messenger; cAMP; Ca2+; signal transduction

资金

  1. JSPS KAKENHI [JP18H02482, JP20K21433, JP21H00435, JP15H05777, JP20K21434]
  2. Japan Science and Technology Agency (JST) Precursory Research for Embryonic Science and Technology (PRESTO) [JPMJPR13A2]
  3. JST Core Research for Evolutional Science and Technology (CREST) [JPMJCR1753]

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Optical control of GPCR signaling is a valuable approach for understanding and controlling GPCR-based physiologies. In this study, the performance of two bistable opsins, MosOpn3 and LamPP, in optical control in vivo was systematically investigated using Caenorhabditis elegans. The results demonstrated the advantages of MosOpn3 and LamPP in light-induced behavior and color-dependent control of behavior, respectively. Molecular engineering was also applied to enhance the usability of these opsins. These findings provide numerous strategies for optical control of GPCR-based physiologies and GPCR signaling.
Optical control of G protein-coupled receptor (GPCR) signaling is a highly valuable approach for comprehensive understanding of GPCR-based physiologies and controlling them precisely. However, optogenetics for GPCR signaling is still developing and requires effective and versatile tools with performance evaluation from their molecular properties. Here, we systematically investigated performance of two bistable opsins that activate Gi/Go-type G protein (mosquito Opn3 (MosOpn3) and lamprey parapinopsin (LamPP)) in optical control in vivo using Caenorhabditis elegans. Transgenic worms expressing MosOpn3, which binds 13-cis retinal to form photopigments, in nociceptor neurons showed light-induced avoidance responses in the presence of all-trans retinal, a retinal isomer ubiquitously present in every tissue, like microbial rhodopsins and unlike canonical vertebrate opsins. Remarkably, transgenic worms expressing MosOpn3 were similar to 7,000 times more sensitive to light than transgenic worms expressing ChR2 in this light-induced behavior, demonstrating the advantage of MosOpn3 as a light switch. LamPP is a UV-sensitive bistable opsin having complete photoregenerative ability by green light. Accordingly, transgenic worms expressing LamPP in cholinergic motor neurons stopped moving upon violet light illumination and restored coordinate movement upon green light illumination, demonstrating color-dependent control of behavior using LamPP. Furthermore, we applied molecular engineering to produce MosOpn3-based tools enabling light-dependent upregulation of cAMP or Ca2+ levels and LamPP-based tool enabling clamping cAMP levels color dependently and context independently, extending their usability. These findings define the capacity of two bistable opsins with similar retinal requirement as ChR2, providing numerous strategies for optical control of various GPCR-based physiologies as well as GPCR signaling itself.

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