Optogenetic induction of hibernation-like state with modified human Opsin4 in mice

Takahashi, Tohru; Hirano, Arisa; Kanda, Takeshi; Saito, Viviane M.; Ashitomi, Hiroto; Tanaka, Kazumasa Z.; Yokoshiki, Yasufumi; Masuda, Kosaku; Yanagisawa, Masashi; Vogt, Kaspar E.; Tokuda, Takashi; Sakurai, Takeshi

doi:10.1016/j.crmeth.2022.100336

Cited by 13 publications

(13 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The QRFP-induced hibernation-like state (QIH) provides a unique model to study metabolic suppression for therapeutic purposes, as well as artificial hibernation. QIH through pharmacogenetics 46 or optogenetics 47 is achievable in murine models, but the prospect of non-genetically triggering these neurons might represent a groundbreaking stride in the realm of synthetic hibernation research. Nevertheless, Q-neurons constitute a remarkably limited subset, and currently there are no established methodologies for their selective pharmacological activation.…”

Section: Discussionmentioning

confidence: 99%

Structure and dynamics of the RF-amide QRFP receptor GPR103

Iwama,

Akasaka,

Sano

et al. 2023

Preprint

View full text Add to dashboard Cite

Pyroglutamylated RF amide peptide (QRFP) is a type of peptide hormone with a C-terminal RF-amide motif. QRFP selectively activates class-A categorized GPCR, GPR103 to exert various physiological functions such as energy metabolism and appetite regulation. Here, we report the cryo-electron microscopy structure of the QRFP-GPR103-Gqcomplex at 3.3 Å resolution. Unlike class-A GPCR, QRFP adopts an extended structure baring no secondary structure, with its N-terminal and C-terminal sides recognized by extracellular and transmembrane domains, respectively, of GPR103. The C-terminal heptapeptide of QRFP penetrates into the orthosteric pocket to act in receptor activation. Particularly, the residues that recognize the RF-amide are highly conserved in the RF-amide receptors. Notably, the unique N-terminal helix-loop-helix of the receptor traps the N-terminal side of QRFP with the pendulum-like motion to guide QRFP into the ligand-binding pocket. This movement, reminiscent of class B1 GPCRs except for orientation and structure of the ligand, is critical for the high affinity binding and receptor specificity of QRFP. Structural comparisons with closely related receptors, including RY-amide peptide-recognizing GPCRs, revealed conserved and diversified peptide recognition mechanisms, providing profound insights into the biological significance of RF-amide peptides. This study not only advances our understanding of GPCR-ligand interactions, but also paves the way for the development of novel therapeutics targeting metabolic and appetite disorders and emergency medical care.

show abstract

Section: Discussionmentioning

confidence: 99%

Structure and dynamics of the RF-amide QRFP receptor GPR103

Iwama,

Akasaka,

Sano

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…The characteristic lower intensity and longer duration light-inducibility is advantageous for in vivo applications, since it will cause less heat generation and less phototoxicity 20,21 . Furthermore, lower intensity light inducibility might presumably be applicable for optogenetic manipulation without intracranial surgery like ChRmine and OPN4dC 22,23 . The LSL-ACR2 mouse strain will provide a new opportunity for researchers to use the e cient inhibitory optogenetic tool.…”

Section: Discussionmentioning

confidence: 99%

Cre-dependent ACR2-expressing reporter mouse strain for efficient long-lasting inhibition of neuronal activity

Mukai

Nakamura

et al. 2023

Preprint

View full text Add to dashboard Cite

Optogenetics is a powerful tool for manipulating neuronal activity by light illumination with high temporal and spatial resolution. Anion-channelrhodopsins (ACRs) are light-gated anion channels that allow researchers to efficiently inhibit neuronal activity. A blue light-sensitive ACR2 has recently been used in several in vivo studies; however, the reporter mouse strain expressing ACR2 has not yet been reported. Here, we generated a new reporter mouse strain, LSL-ACR2, in which ACR2 is expressed under the control of Cre recombinase. We crossed this strain with a noradrenergic neuron-specific driver mouse (NAT-Cre) to generate NAT-ACR2 mice. We confirmed Cre-dependent expression and function of ACR2 in the targeted neurons by immunohistochemistry and electrophysiological recordings in vitro, and confirmed physiological function using an in vivo behavioral experiment. Our results show that the LSL-ACR2 mouse strain can be applied for optogenetic inhibition of targeted neurons, particularly for long-lasting continuous inhibition, upon crossing with Cre-driver mouse strains. The LSL-ACR2 strain can be used to prepare transgenic mice with homogenous expression of ACR2 in targeted neurons with a high penetration ratio, good reproducibility, and no tissue invasion.

show abstract

“…QIH through pharmacogenetics 42 or optogenetics 43 is achievable in murine models, but the prospect of non-genetically triggering these neurons might represent a groundbreaking stride in the realm of synthetic hibernation research. Nevertheless, Q-neurons constitute a remarkably limited subset, and currently there are no established methodologies for their selective pharmacological activation.…”

Section: Discussionmentioning

confidence: 99%

Structure and dynamics of the RF-amide QRFP receptor GPR103

Nureki,

Iwama,

Akasaka

et al. 2023

Preprint

View full text Add to dashboard Cite

Pyroglutamylated RF amide peptide (QRFP) is a type of peptide hormone with a C-terminal RF-amide motif. QRFP selectively activates class-A categorized GPCR, GPR103 to exert various physiological functions such as energy metabolism and appetite regulation. Here, we report the cryo-electron microscopy structure of the QRFP-GPR103-Gq complex at 3.3 Å resolution. Unlike class-A GPCR, QRFP adopts an extended structure baring no secondary structure, with its N-terminal and C-terminal sides recognized by extracellular and transmembrane domains, respectively, of GPR103. The C-terminal heptapeptide of QRFP penetrates into the orthosteric pocket to act in receptor activation. Particularly, the residues that recognize the RF-amide are highly conserved in the RF-amide receptors. Notably, the unique N-terminal helix-loop-helix of the receptor traps the N-terminal side of QRFP with the pendulum-like motion to guide QRFP into the ligand-binding pocket. This movement, reminiscent of class B1 GPCRs except for orientation and structure of the ligand, is critical for the high affinity binding and receptor specificity of QRFP. Structural comparisons with closely related receptors, including RY-amide peptide-recognizing GPCRs, revealed conserved and diversified peptide recognition mechanisms, providing profound insights into the biological significance of RF-amide peptides. This study not only advances our understanding of GPCR-ligand interactions, but also paves the way for the development of novel therapeutics targeting metabolic and appetite disorders and emergency medical care.

show abstract

Optogenetic induction of hibernation-like state with modified human Opsin4 in mice

Cited by 13 publications

References 67 publications

Structure and dynamics of the RF-amide QRFP receptor GPR103

Structure and dynamics of the RF-amide QRFP receptor GPR103

Cre-dependent ACR2-expressing reporter mouse strain for efficient long-lasting inhibition of neuronal activity

Structure and dynamics of the RF-amide QRFP receptor GPR103

Contact Info

Product

Resources

About