Characterization of human Neuropsin (hOPN5) and development of new mutants for optogenetic stimulation of the Gq signaling cascade DOI Creative Commons

Udhaya Bhaskar Sathya Narayanan

Published: Jan. 1, 2024

Gq signaling plays a pivotal role in the physiological adaptation of cellular behavior based on individual body's needs and it is also involved pathogenesis many diseases including cancer, autoimmunity, pathological cardiac hypertrophy, heart failure. However, transition between underlying kinetics not well understood since current approaches with pharmacological stimulation genetic modulation lack spatiotemporal precision. In striking contrast, optogenetic G protein-coupled receptor (GPCR) offers an unprecedented advantage high temporal spatial precision chance cell-specific expression. Therefore, GPCR would be ideal tool to study transitions kinetics. this thesis, I characterized wild-type (WT) Neuropsin (OPN5), ultra-violet (UV) light-sensitive bistable GPCR, prove its selective activation pathway heterologous expression systems transgenic mouse model. UV light-induced Inositol 1,4,5 trisphosphate (IP3) production as transient potential canonical cation channel 6 (TRPC6) channels by Diacylglycerol (DAG) was demonstrated HEK293 hOPN5 cells. Using Ca2+ imaging, red-shifted Xanthene-based rhodamine like fluorescent indicator-1 (X-Rhod-1, 1.5 µM), light sensitivity (3.26 ± 0.17 µW/mm2, n=17) most efficient wavelength for (388 2.7 nm, n=14) were determined (WT). further compared proved that light-stimulated transients abolished Gq/11 knock-out (KO) cells while preserved Gi KO protein activated inward rectifying K+ (GIRK) assay excluded promiscuous coupling By combining structural modeling sequence alignments, few mutations predicted around 8 Å region retinal chromophore alter photo-excited state, thereby having fine-tuned spectrum altered The rate strength mutant along chicken OPN5 (cOPN5) analyzed. They using imaging. sensitivities ranging from (eLi50 µW/mm2) 160 2. shifted maximally 388 (hOPN5 (WT), n=10) 406 2.5 (cOPN5, n=10). Investigation dependence among OPN5s hinted at importance threonine alanine mutation. Taken together, control cells, tissues, organs. Further, development new mutants which are expected have spectral properties pave way foundation better understand important amino acid positions aid Molecular dynamics (MD) simulations. Finally, goal develop enhanced highly OPN5-based tools future organ function.

Language: Английский

Direct photoreception by pituitary endocrine cells regulates hormone release and pigmentation DOI
Ayaka Fukuda, Keita Sato, Chika Fujimori

et al.

Science, Journal Year: 2025, Volume and Issue: 387(6729), P. 43 - 48

Published: Jan. 2, 2025

The recent discovery of nonvisual photoreceptors in various organs has raised expectations for uncovering their roles and underlying mechanisms. In this work, we identified a previously unrecognized hormone-releasing mechanism the pituitary Japanese rice fish (medaka) induced by light. Ca 2+ imaging analysis revealed that melanotrophs, type endocrine cell secretes melanocyte-stimulating hormone, robustly increase concentration intracellular during short-wavelength light exposure. Moreover, Opn5m as key molecule drives response. Knocking out opn5m attenuated melanogenesis reducing tyrosinase expression skin. Our findings suggest which direct reception melanotrophs triggers pathway might contribute to protection from ultraviolet radiation medaka.

Language: Английский

Citations

2
An Expanding Role for Nonvisual Opsins in Extraocular Light Sensing Physiology DOI Creative Commons

Mutahar Andrabi,

Brian A. Upton, Richard A. Lang

et al.

Annual Review of Vision Science, Journal Year: 2023, Volume and Issue: 9(1), P. 245 - 267

Published: May 17, 2023

We live on a planet that is bathed in daily and seasonal sunlight cycles. In this context, terrestrial life forms have evolved mechanisms directly harness light energy (plants) or decode information for adaptive advantage. animals, the main sensors are family of G protein-coupled receptors called opsins. Opsin function best described visual sense. However, most animals also use opsins extraocular sensing behavior camouflage. While it has long been believed mammals do not an capacity, recent evidence suggests otherwise. Notably, encephalopsin (OPN3) neuropsin (OPN5) both known to mediate mice. Examples mediation include photoentrainment circadian clocks skin (by OPN5) acute light-dependent regulation metabolic pathways OPN3 OPN5). This review summarizes current findings expanding field photoreception their relevance human physiology.

Language: Английский

Citations

28
A bistable inhibitory optoGPCR for multiplexed optogenetic control of neural circuits DOI Creative Commons
Jonas Wietek,

Adrianna Nozownik,

Mauro Pulin

et al.

Nature Methods, Journal Year: 2024, Volume and Issue: 21(7), P. 1275 - 1287

Published: May 29, 2024

Abstract Information is transmitted between brain regions through the release of neurotransmitters from long-range projecting axons. Understanding how activity such connections contributes to behavior requires efficient methods for reversibly manipulating their function. Chemogenetic and optogenetic tools, acting endogenous G-protein-coupled receptor pathways, can be used modulate synaptic transmission, but existing tools are limited in sensitivity, spatiotemporal precision or spectral multiplexing capabilities. Here we systematically evaluated multiple bistable opsins applications found that Platynereis dumerilii ciliary opsin ( Pd CO) an efficient, versatile, light-activated suppress transmission mammalian neurons with high temporal vivo. CO has useful biophysical properties enable other actuators reporters. We demonstrate conduct reversible loss-of-function experiments projections behaving animals, thereby enabling detailed synapse-specific functional circuit mapping.

Language: Английский

Citations

15
Multiple opsins in a reef-building coral, Acropora millepora DOI Creative Commons

Benjamin Mason,

Mitsumasa Koyanagi,

Tomohiro Sugihara

et al.

Scientific Reports, Journal Year: 2023, Volume and Issue: 13(1)

Published: Jan. 29, 2023

Abstract Opsins, light-sensitive G protein-coupled receptors, have been identified in corals but their properties are largely unknown. Here, we six opsin genes (acropsins 1–6) from a coral species Acropora millepora , including three novel opsins 4–6), and successfully characterized the of four out acropsins. Acropsins 1 6 exhibited light-dependent cAMP increases cultured cells, suggesting that acropsins could light-dependently activate Gs-type protein like box jellyfish same group. Spectral sensitivity curves having maximum sensitivities at ~ 472 nm 476 were estimated for 6, respectively, based on light wavelength-dependent these opsins-expressing cells (heterologous action spectroscopy). Acropsin 2 belonging to group as did not induce or Ca 2+ changes. We then acropsin spectral curve its value 471 with chimera mutant which possessed third cytoplasmic loop Gs-coupled opsin. 4 categorized another induced intracellular Our results uncovered possesses multiple coupling two distinct cascades, cyclic nucleotide signaling light-dependently.

Language: Английский

Citations

17
Molecular Mechanisms of the Melatonin Receptor Pathway Linking Circadian Rhythm to Type 2 Diabetes Mellitus DOI Open Access

Anyu Xia,

Hui Zhu,

Zhijia Zhao

et al.

Nutrients, Journal Year: 2023, Volume and Issue: 15(6), P. 1406 - 1406

Published: March 15, 2023

Night-shift work and sleep disorders are associated with type 2 diabetes (T2DM), circadian rhythm disruption is intrinsically involved. Studies have identified several signaling pathways that separately link two melatonin receptors (MT1 MT2) to insulin secretion T2DM occurrence, but a comprehensive explanation of the molecular mechanism elucidate association between these T2DM, reasonably precisely, has been lacking. This review thoroughly explicates system, which consists four important pathways, linking MT1 or MT2 secretion. Then, MTNR1B transcription extensively expounded. Finally, concrete evolutionary underlying macroscopic established. provides new insights into pathology, treatment, prevention T2DM.

Language: Английский

Citations

14
Optogenetic engineering for ion channel modulation DOI
Tianlu Wang, Teruo Nonomura,

Tien‐Hung Lan

et al.

Current Opinion in Chemical Biology, Journal Year: 2025, Volume and Issue: 85, P. 102569 - 102569

Published: Feb. 3, 2025

Language: Английский

Citations

0
Characterization of Anthozoan-Specific Opsins from a Reef-Building Coral, Acropora tenuis, as Gq-Coupled Opsins DOI
Makoto Iwasaki, Yusuke Sakai,

Taishi Shirata

et al.

ZOOLOGICAL SCIENCE, Journal Year: 2025, Volume and Issue: 42(2)

Published: March 19, 2025

Most animals capture light through opsins, which are light-sensitive G protein-coupled receptors (GPCRs). Recent genome analyses of anthozoans, including corals and sea anemones, have identified novel opsins that phylogenetically classified into two groups distinct from previously known opsin groups. Despite their significance in clarifying biological functions, the specific molecular properties these remain largely unknown. In this study, we investigated protein activations biochemical responses light-dependently induced by anthozoan Antho2a Antho2d, obtained reef-building coral Acropora tenuis, mammalian cultured cells. Using jumping spider Rh1 (SpiRh1), belongs to a Gq-type (Gq)-coupled group as control, observed Antho2d elicited light-dependent increases Ca2+ levels This response was inhibited Gq inhibitor, indicating activated manner. Interestingly, also Gi-type (Gi), similar SpiRh1, while showed limited or negligible Gi activation. We found activation additionally contributed elevation, suggesting it enhances Gq-dependent elevation Antho2d- SpiRh1-expressing contrast, demonstrated higher specificity for compared SpiRh1 nearly equivalent hM3Dq, GPCR its strong widely used chemogenetic tool manipulating Our results suggest new consists Gq-coupled with varying activation, demonstrating potential optogenetic applications.

Language: Английский

Citations

0
Direct Stimulation of Gastric Smooth Muscle Cells via Gq Proteins With Light DOI Creative Commons

David Zipf,

Markus Vogt, Udhayabhaskar Sathyanarayanan

et al.

Neurogastroenterology & Motility, Journal Year: 2025, Volume and Issue: unknown

Published: March 31, 2025

ABSTRACT Background Optogenetics is a cutting‐edge approach that can enable direct stimulation of gastric smooth muscle cells (SMC) by combining cell‐specific overexpression light‐sensitive proteins with light stimulation. We previously demonstrated optogenetic SMC via depolarization restore contractility and food propulsion could become new treatment strategy for gastroparesis. The human receptor Neuropsin (hOPN5) enables activation G q signaling UV light. Herein, we explore this SMC. Methods used transgenic mouse model expressing hOPN5 in fusion eYFP. Antral longitudinal strips were isometric force measurements whole stomachs intragastric pressure measurements, comparing to other stimuli. Adeno‐associated virus (AAV) serotypes screened efficiency transducing cultured SMC, transduced tested Ca 2+ imaging. Results expression was restricted found ~1/3 the stomach. induced increased only mice similarly electrical field reached approximately 1/3 global muscarinic activation. Importantly, optical remained effective an ex vivo gastroparesis model. AAV 2.5 far most serotype transduction, triggered transients hOPN5. Conclusion tool directly stimulate control Thus, it additional complementary light‐induced membrane motility.

Language: Английский

Citations

0
A neuropsin-based optogenetic tool for precise control of Gq signaling DOI

Ruicheng Dai,

Yu Tao,

Danwei Weng

et al.

Science China Life Sciences, Journal Year: 2022, Volume and Issue: 65(7), P. 1271 - 1284

Published: May 12, 2022

Language: Английский

Citations

16
Emerging molecular technologies for light-mediated modulation of pancreatic beta-cell function DOI Creative Commons
Zijing Chen,

Leah F. Truskinovsky,

Emmanuel S. Tzanakakis

et al.

Molecular Metabolism, Journal Year: 2022, Volume and Issue: 64, P. 101552 - 101552

Published: July 19, 2022

Optogenetic modalities as well optochemical and photopharmacological strategies, collectively termed optical methods, have revolutionized the control of cellular functions via light with great spatiotemporal precision. In comparison to major advances in photomodulation signaling activities noted neuroscience, similar applications endocrine cells pancreas, particularly insulin-producing β-cells, been limited. The availability tools allowing light-mediated changes trafficking ions such K+ Ca2+ intermediates cyclic adenosine monophosphate (cAMP), renders β-cells their glucose-stimulated insulin secretion (GSIS) amenable optoengineering for drug-free blood sugar. molecular circuit GSIS is described emphasis on which are targetable intervention. Various pharmacological agents modifying release reviewed along documented side effects. These contrasted approaches, already employed engineering β-cell function or considered future applications. Principal obstacles also discussed implementation optogenetics pondered tissue biology pancreas. Notable optogenetic, rendering feasible smart pancreatic tissues light-regulated paving way novel solutions addressing pathologies including diabetes.

Language: Английский

Citations

16