Rapid and reversible optogenetic silencing of synaptic transmission by clustering of synaptic vesicles DOI Creative Commons

Dennis Vettkötter,

Martin Schneider, Brady D. Goulden

и другие.

Nature Communications, Год журнала: 2022, Номер 13(1)

Опубликована: Дек. 19, 2022

Abstract Acutely silencing specific neurons informs about their functional roles in circuits and behavior. Existing optogenetic silencers include ion pumps, channels, metabotropic receptors, tools that damage the neurotransmitter release machinery. While former hyperpolarize cell, alter ionic gradients or cellular biochemistry, latter allow only slow recovery, requiring de novo synthesis. Thus, combining fast activation reversibility are needed. Here, we use light-evoked homo-oligomerization of cryptochrome CRY2 to silence synaptic transmission, by clustering vesicles (SVs). We benchmark this tool, optoSynC, Caenorhabditis elegans , zebrafish, murine hippocampal neurons. optoSynC clusters SVs, observable electron microscopy. Locomotion occurs with tau on ~7.2 s recovers off ~6.5 min after light-off. can inhibit exocytosis for several hours, at very low light intensities, does not affect currents, biochemistry proteins, may further manipulating different SV pools transfer SVs between them.

Язык: Английский

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

Adrianna Nozownik,

Mauro Pulin

и другие.

Nature Methods, Год журнала: 2024, Номер 21(7), С. 1275 - 1287

Опубликована: Май 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.

Язык: Английский

Процитировано

15

Neuromodulator and neuropeptide sensors and probes for precise circuit interrogation in vivo DOI
Jessie Muir, Maribel Anguiano, Christina K. Kim

и другие.

Science, Год журнала: 2024, Номер 385(6716)

Опубликована: Сен. 26, 2024

To determine how neuronal circuits encode and drive behavior, it is often necessary to measure manipulate different aspects of neurochemical signaling in awake animals. Optogenetics calcium sensors have paved the way for these types studies, allowing perturbation readout spiking activity within genetically defined cell types. However, methods lack ability further disentangle roles individual neuromodulator neuropeptides on behavior. We review recent advances chemical biology tools that enable precise spatiotemporal monitoring control over neuroeffectors their receptors vivo. also highlight discoveries enabled by such tools, revealing molecules signal across timescales learning, orchestrate behavioral changes, modulate circuit activity.

Язык: Английский

Процитировано

12

Rhodopsins at a glance DOI Open Access
Takashi Nagata, Keiichi Inoue

Journal of Cell Science, Год журнала: 2021, Номер 134(22)

Опубликована: Ноя. 15, 2021

Rhodopsins are photoreceptive membrane proteins consisting of a common heptahelical transmembrane architecture that contains retinal chromophore. Rhodopsin was first discovered in the animal retina 1876, but different type rhodopsin, bacteriorhodopsin, reported to be present cell an extreme halophilic archaeon, Halobacterium salinarum, 95 years later. Although these findings were made by physiological observation pigmented tissue and bodies, recent progress genomic metagenomic analyses has revealed there more than 10,000 microbial rhodopsins 9000 with large diversity tremendous new functionality. In this Cell Science at Glance article accompanying poster, we provide overview functions, structures, color discrimination mechanisms optogenetic applications two rhodopsin families, will also highlight third distinctive family, heliorhodopsin.

Язык: Английский

Процитировано

47

Advancing respiratory–cardiovascular physiology with the working heart–brainstem preparation over 25 years DOI Creative Commons
Julian F. R. Paton, Benedito H. Machado, Davi J. A. Moraes

и другие.

The Journal of Physiology, Год журнала: 2022, Номер 600(9), С. 2049 - 2075

Опубликована: Март 16, 2022

Abstract Twenty‐five years ago, a new physiological preparation called the working heart–brainstem (WHBP) was introduced with claim it would provide platform allowing studies not possible before in cardiovascular, neuroendocrine, autonomic and respiratory research. Herein, we review some of progress made WHBP, advantages disadvantages along potential future applications, photographs technical drawings all customised equipment used for preparation. Using mice or rats, WHBP is an situ experimental model that perfused via extracorporeal circuit benefitting from unprecedented surgical access, mechanical stability brain whole cell recording uncompromised use pharmacological agents akin to vitro approaches. The has revealed novel mechanistic insights into, example, generation distinct rhythms, neurogenesis sympathetic activity, coupling between respiration heart circulation, hypothalamic spinal control mechanisms, peripheral central chemoreceptor mechanisms. Insights have been gleaned into diseases such as hypertension, failure sleep apnoea. Findings ratified conscious vivo animals when tested translated humans. We conclude by discussing applications including two‐photon imaging nervous systems adoption pharmacogenetic tools will improve our understanding mechanisms reveal may guide treatment strategies cardiorespiratory diseases. image

Язык: Английский

Процитировано

38

Wireless, Battery-Free Implants for Electrochemical Catecholamine Sensing and Optogenetic Stimulation DOI
Tucker Stuart, William J. Jeang, Richard A. Slivicki

и другие.

ACS Nano, Год журнала: 2022, Номер 17(1), С. 561 - 574

Опубликована: Дек. 22, 2022

Neurotransmitters and neuromodulators mediate communication between neurons other cell types; knowledge of release dynamics is critical to understanding their physiological role in normal pathological brain function. Investigation into transient neurotransmitter has largely been hindered due electrical material requirements for electrochemical stimulation recording. Current systems require complex electronics biasing amplification rely on materials that offer limited sensor selectivity sensitivity. These restrictions result bulky, tethered, or battery-powered impacting behavior constant care subjects. To overcome these challenges, we demonstrate a fully implantable, wireless, battery-free platform enables optogenetic recording catecholamine real time. The device nearly 1/10th the size previously reported examples includes probe relies multilayer electrode architecture featuring microscale light emitting diode (μ-LED) carbon nanotube (CNT)-based with sensitivities among highest recorded literature (1264.1 nA μM

Язык: Английский

Процитировано

35

G protein gamma subunit, a hidden master regulator of GPCR signaling DOI Creative Commons
Dinesh Kankanamge, Mithila Tennakoon,

Ajith Karunarathne

и другие.

Journal of Biological Chemistry, Год журнала: 2022, Номер 298(12), С. 102618 - 102618

Опубликована: Окт. 19, 2022

Heterotrimeric G proteins (αβγ subunits) that are activated by protein-coupled receptors (GPCRs) mediate the biological responses of eukaryotic cells to extracellular signals. The α subunits and tightly bound βγ subunit complex have been extensively studied shown control activity effector molecules. In contrast, potential roles large family γ less studied. this review, we focus on present knowledge about these proteins. Induced loss individual types in animal plant models result strikingly distinct phenotypes indicating subtypes play important specific roles. Consistent with findings, downregulation or upregulation particular various cancers. Clues mechanistic basis function emerged from imaging dynamic behavior protein living cells. This shows basal state, not constrained plasma membrane but shuttle between membranes receptor activation complexes translocate reversibly internal membranes. translocation kinetics varies widely is determined affinity associated subtype. On translocating, some act effectors variation determines differential sensitivity adaptation external Membrane thus a parsimonious elegant mechanism controls information flow cell while modulating signaling responses. sense signals activate heterotrimeric (αβγ) Activation results exchanging GDP for GTP dissociation complex. Both α-GTP independently capable effectors. GTPase switches active GTP-bound form deactivated (1Gilman A.G. proteins: transducers receptor-generated signals.Annu. Rev. Biochem. 1987; 56: 615-649Crossref PubMed Google Scholar). families diverse members number such as adenylyl cyclase phospholipase C (2Simon M.I. Strathmann M.P. Gautam N. Diversity signal transduction.Science. 1991; 252: 802-808Crossref Scholar, 3Gautam Downes G.B. Yan K. Kisselev O. G-protein betagamma complex.Cell Signal. 1998; 10: 447-455Crossref Scopus (158) acts including protein-gated inwardly rectifying K+ channels (4Logothetis D.E. Kurachi Y. Galper J. Neer E.J. Clapham beta gamma GTP-binding muscarinic channel heart.Nature. 325: 321-326Crossref (876) Scholar), (5Tang W.J. Gilman Type-specific regulation subunits.Science. 254: 1500-1503Crossref (PLC) (6Katz A. Wu D. Simon Subunits 2 isoform C.Nature. 1992; 360: 686-689Crossref (0) GPCR kinases (7Daaka Pitcher J.A. Richardson M. Stoffel R.H. Robishaw J.D. Lefkowitz R.J. Receptor isoform-specific interactions kinases.Proc. Natl. Acad. Sci. U. S. 1997; 94: 2180-2185Crossref (155) phosphoinositide 3-kinase (PI3Kγ) (8Brock C. Schaefer Reusch H.P. Czupalla Michalke Spicher et al.Roles recruitment p110 gamma/p101 gamma.J. Cell Biol. 2003; 160: 89-99Crossref Recent reviews focused aspects (9Hewavitharana T. Wedegaertner P.B. Non-canonical localizations proteins.Cell 2012; 24: 25-34Crossref (71) 10Campbell A.P. Smrcka A.V. Targeting signalling blocking proteins.Nat. Drug Discov. 2018; 17: 789-803Crossref (76) 11Tennakoon Senarath Kankanamge Ratnayake Wijayaratna Olupothage al.Subtype-dependent Gbetagamma signalling.Cell 2021; 82: 109947Crossref studies limited their structure functions reviewed. review focuses subunits, based information, gaps remain our knowledge, future experimental directions can address lacunae. transducin, found rod outer segments retina was first be characterized at cDNA level (12Hurley J.B. Fong H.K. Teplow D.B. Dreyer Isolation characterization clone bovine retinal transducin.Proc. 1984; 81: 6948-6952Crossref identification Gi/o using peptide analysis PCR showed primary structures two diverged considerably, it evolutionarily related small binding Ras (13Gautam Baetscher Aebersold R. A shares homology ras proteins.Science. 1989; 244: 971-974Crossref Identification additional suggested were potentially structurally (14Gautam Northup Tamir H. diversity increased associations variety subunits.Proc. 1990; 87: 7973-7977Crossref Over years, 12 identified sequences 15Fisher K.J. Aronson Jr., N.N. Characterization genomic sequence (gamma 5).Mol. 12: 1585-1591Crossref 16Kalyanaraman Kalyanaraman V. brain-specific subunit.Biochem. Biophys. Res. Commun. 1995; 216: 126-132Crossref 17Ray Kunsch Bonner L.M. clones encoding eight different human three novel forms designated 4, 10, 11 subunits.J. Chem. 270: 21765-21771Abstract Full Text PDF 18Morishita Nakayama Isobe Matsuda Hashimoto Okano al.Primary protein, 12, its phosphorylation kinase C.J. 29469-29475Abstract (79) 19Ryba N.J. Tirindelli gamma-subunit, 8, expressed during neurogenesis olfactory vomeronasal neuroepithelia.J. 6757-6767Abstract 20Downes gene families.Genomics. 1999; 62: 544-552Crossref (228) 21Huang L. Shanker Y.G. Dubauskaite Zheng J.Z. W. Rosenzweig al.Ggamma13 colocalizes gustducin taste mediates IP3 bitter denatonium.Nat. Neurosci. 2: 1055-1062Crossref (285) conserved mammalian species differences amino acid residues among functional importance (3Gautam presence yeast (22Whiteway Hougan Dignard Thomas D.Y. Bell Saari G.C. al.The STE4 STE18 genes encode mating factor receptor-coupled protein.Cell. 467-477Abstract Scholar) plants (23Mason M.G. Botella J.R. Completing heterotrimer: isolation an Arabidopsis thaliana gamma-subunit cDNA.Proc. 2000; 97: 14784-14788Crossref 24Mason interaction Gbeta.Biochim. Acta. 2001; 1520: 147-153Crossref also has retained over long period evolution all eukaryotes further emphasized independent role signaling. thaliana, atypical distinctly other (25Chakravorty Trusov Zhang Acharya B.R. Sheahan M.B. McCurdy D.W. al.An involved guard K(+)-channel morphological development thaliana.Plant 2011; 67: 840-851Crossref (167) suggests considerably specialized Though lipidation covalent 16-carbon palmitate group and/or 14 carbon myristate N terminus discovered late 1980s (26Wedegaertner Wilson P.T. Bourne H.R. Lipid modifications trimeric proteins.J. 503-506Abstract (394) only 1990 anchoring via prenyl moiety posttranslational (27Fukada Takao Ohguro Yoshizawa Akino Shimonishi Farnesylated photoreceptor indispensable GTP-binding.Nature. 346: 658-660Crossref 28Xie Yamane Stephenson Ong Fung B. Clarke Analysis prenylated carboxyl-terminal cysteine methyl esters proteins.Methods. 1: 276-282Crossref (17) 29Chen C.A. Manning D.R. Regulation modification.Oncogene. 20: 1643-1652Crossref (169) lipidated group, either farnesyl (15 Carbon) 30Lai R.K. Perez-Sala Canada F.J. Rando R.R. transducin farnesylated.Proc. 7673-7677Crossref geranylgeranyl (20 (31Mumby S.M. Casey P.J. Gutowski Sternweis P.C. contain 20-carbon isoprenoid.Proc. 5873-5877Crossref 32Yamane Farnsworth C.C. Xie H.Y. Howald B.K. al.Brain all-trans-geranylgeranylcysteine ester carboxyl termini.Proc. 5868-5872Crossref through stable thioether linkage C-terminal Cys. four-residue called; "the CaaX motif" type prenylation composed Cys, aliphatic acids-aa, transferase determining residue, X. Cys farnesylated when X Met, Ser, Glu, Ala (as γ1, γ9, γ11), geranylgeranylated Leu (the rest nine (33Vogler Barcelo J.M. Ribas Escriba P.V. proteins.Biochim. 2008; 1778: 1640-1652Crossref last (aaX) proteolytically cleaved off endoprotease; converting endopeptidase, subsequently carboxy methylated methyltransferase, isoprenyl-cysteine 34Gao Liao Yang G.Y. CAAX-box process carcinogenesis.Am. Transl 2009; 312-325PubMed contrast which restricted set life modified proteins, ubiquitous transient. Phosphorylation occur case γ12, pathways (18Morishita 35Yasuda Lindorfer M.A. Myung C.S. Garrison J.C. gamma12 regulates specificity.J. 273: 21958-21965Abstract (45) 36Ueda Yamauchi Itoh Morishita Kaziro Kato al.Phosphorylation F-actin-associating enhances fibroblast motility.J. 274: 12124-12128Abstract More recently, essential downstream (37Nassiri Toosi Z. Su Austin Choudhury Li Pang Y.T. al.Combinatorial modulates yeast.Sci. 14eabd2464Crossref (3) An examination putative sites N-terminal (38Chakravorty Assmann regulatory mammals, yeast, plants.Biochem. 475: 3331-3357Crossref will become clearer whether phosphorylated, general theme regulating subunits. Since there possibility made up combinations β could functions, determine rules association types. Do associate selective association? Such selectivity would suggest even if expresses many types, certain possible. methods (39Pronin A.N. Interaction selective.Proc. 89: 6220-6224Crossref 40Schmidt T.C. Levine Specificity interactions.J. 267: 13807-13810Abstract 41Yan Differential ability families.J. 1996; 271: 7141-7146Abstract (84) 42Dingus Wells Campbell Cleator J.H. Robinson Hildebrandt Protein dimer formation: gbeta Ggamma differentially efficiency vitro formation.Biochemistry. 2005; 44: 11882-11890Crossref Importantly, purifying native tissues confirmed (43Asano Ueda Selective beta(4) gamma(5) gamma(12) tissues.J. 21425-21429Abstract When heterotrimers identity examined, they suggesting again 44Hildebrandt Codina Rosenthal Birnbaumer Yamazaki al.Characterization two-dimensional mapping Ns Ni, cyclase, guanine nucleotide-binding eye.J. 1985; 260: 14867-14872Abstract 45Tamir Fawzi A.B. Evans J.K. G-Protein forms: subunits.Biochemistry. 30: 3929-3936Crossref 46Wilcox M.D. Dingus Balcueva E.A. McIntire W.E. Mehta N.D. Schey K.L. al.Bovine brain GO isoforms compositions.J. 4189-4192Abstract 47Richardson alpha2A-adrenergic discriminates Gi composition Sf9 insect membranes.J. 13525-13533Abstract (75) Knocking down line antisense oligonucleotides provided support selectively affecting (48Kleuss Scherubl Hescheler Schultz G. Wittig Selectivity transduction 1993; 259: 832-834Crossref After early mouse elucidation (49Downes Copeland N.G. Jenkins N.A. Structure gamma3 divergently transcribed gene, gng3lg.Genomics. 53: 220-230Crossref genomics now comprehensive both (Tables 1 2). earlier γ1 γ11 closely arranged together head tail orientation may arisen duplication, γ3 Gng3lg (20Downes later named humans BSCL2, mutations congenital lipodystrophy, Berardinelli–Seip syndrome (50Magre Delepine Khallouf E. Gedde-Dahl Van Maldergem Sobel al.Identification altered Berardinelli-Seip lipodystrophy chromosome 11q13.Nat. Genet. 28: 365-370Crossref (567) Scholar).Table 1Mouse genesaData adapted database resources National Center Biotechnology Information.Gene symbolGene idChromosome noNumber exonsGNG114,69966GNG214,702148GNG314,704193GNG414,706135GNG514,70733GNG714,708107GNG814,70976GNG914,710117GNG1014,70043GNG1166,06662GNG1214,70166GNG1364,337174a Data Information. Open table new tab Table 2Human Gγ exonsGNG1279273GNG254,3311414GNG32785115GNG4278618GNG5278714GNG72788196GNG894,235195GNG92793175GNG10279093GNG11279172GNG1255,97017GNG1351,764163a There suggestions direct receptors. Studies purified obligatory requirement 51Birnbaumer transduction.Annu. Pharmacol. Toxicol. 675-705Crossref activation. Peptides domain stabilized photoactivated rhodopsin, region prevented heterotrimer rhodopsin (52Kisselev O.G. Ermolaeva M.V. determinant coupling.J. 1994; 269: 21399-21402Abstract 53Kisselev Pronin Receptor-G coupling established conformational switch beta-gamma complex.Proc. 92: 9102-9106Crossref results, change light-activated detected same remained disordered inactive dark-adapted (54Kisselev Downs Rhodopsin subunit.Structure. 11: 367-373Abstract (62) corresponding γ5 subunit, γ7 γ12 inhibit M2 signaling, Gγ-receptor (55Azpiazu I. Cruzblanca P. Linder Zhuo subunit-specific inhibits signaling.J. 35305-35308Abstract supported findings more potent supporting (47Richardson 55Azpiazu 56Yasuda Woodfork K.A. Fletcher J.E. Role A1 adenosine receptors.J. 18588-18595Abstract 57Hou Azpiazu interaction.J. 275: 38961-38964Abstract 58Lim W.K. Neubig specificity: gamma11 unique potency A(1) 5-HT(1A) receptors.Biochemistry. 40: 10532-10541Crossref 20 available receptor–G heterotrimer, them containing γ2 without site. Their PDB IDs (Table 3). complete clear any likely due hypervariable nature domain. GPCR-G capture frozen states narrow time window, possible captured occurs. notion, recent modeling how existent fit into model where occurs transiently intracellular hydrophobic site facilitating subsequent (59McIntire couples Galpha GPCR.J. Gen. Physiol. 2022; 154e202112982Crossref (1) Structures transient after directly questions receptor.Table 3Cryo-EM X-ray crystallographic complexesaStructural data Bank.a Structural Bank. Once family, expression examined 4). tissue. antisera used, extracts These γ5, γ10, several tissues, although γ10 barely detectable (60Cali J.J. Rybalkin tissue distribution cloning.J. 24023-24027Abstract 61Asano Ohashi Nagahama Miyake Localization neural nonneural Neurochem. 64: 1267-1273Crossref 62Morishita Asano gamma10 gamma11, lung rat.FEBS Lett. 428: 85-88Crossref indication roles.Table 4Genomic location tissue-specific subunitsaGenomic adapt

Язык: Английский

Процитировано

34

Neurophotonic Tools for Microscopic Measurements and Manipulation: Status Report DOI Creative Commons
Ahmed S. Abdelfattah,

Srinivasa Rao Allu,

Robert E. Campbell

и другие.

Neurophotonics, Год журнала: 2022, Номер 9(S1)

Опубликована: Апрель 27, 2022

Neurophotonics was launched in 2014 coinciding with the launch of BRAIN Initiative focused on development technologies for advancement neuroscience. For last seven years, Neurophotonics' agenda has been well aligned this focus neurotechnologies featuring new optical methods and tools applicable to brain studies. While 2.0 is pivoting towards applications these novel quest understand brain, article we review an extensive diverse toolkit explore function that have emerged from related large-scale efforts measurement manipulation structure function. Here, neurophotonic mostly animal A companion article, scheduled appear later year, will cover diffuse imaging noninvasive human each domain, outline current state-of-the-art respective technologies, identify areas where innovation needed provide outlook future directions.

Язык: Английский

Процитировано

33

Spatiotemporally Precise Optical Manipulation of Intracellular Molecular Activities DOI Creative Commons
Bin Dong, Shivam Mahapatra,

Matthew G. Clark

и другие.

Advanced Science, Год журнала: 2024, Номер 11(13)

Опубликована: Янв. 26, 2024

Abstract Controlling chemical processes in live cells is a challenging task. The spatial heterogeneity of biochemical reactions often overlooked by conventional means incubating with desired chemicals. A comprehensive understanding spatially diverse requires precise control over molecular activities at the subcellular level. Herein, closed‐loop optoelectronic system developed that allows manipulation biomolecular high spatiotemporal precision. Chemical‐selective fluorescence signals are utilized to command lasers trigger specific or activation photoswitchable inhibitors targets. This technology fully compatible laser scanning confocal microscopes. authors demonstrate selective interactions 405 nm targeted organelles and simultaneous monitoring cell responses fluorescent protein signals. Notably, blue interaction endoplasmic reticulum leads more pronounced reduction cytosolic green comparison nuclei lipid droplets. Moreover, when combined inhibitor, microtubule polymerization selectively inhibited within compartments. enables optical drug activities, exclusively targets, while minimizing undesired effects on non‐targeted locations.

Язык: Английский

Процитировано

8

Characterization and Engineering of a Blue-Sensitive, Gi/o-Biased, and Bistable Ciliary Opsin from a Fan Worm DOI
Seketsu Fukuzawa, Takashi Kawaguchi, Takushi Shimomura

и другие.

Biochemistry, Год журнала: 2025, Номер unknown

Опубликована: Фев. 13, 2025

Ciliary opsins (c-opsin) have been identified not only in vertebrates but also invertebrates. An invertebrate ciliary opsin was recently the fan worm Acromegalomma interruptum (formerly named Megalomma interrupta); however, its spectral and signaling characteristics are unknown. In present study, we characterized properties light-induced cellular of (Acromegalomma (AcrInvC-opsin)). AcrInvC-opsin showed an absorption maximum at 464 nm, upon blue light absorption, spectrum red-shifted by approximately 50 nm. The two states interconvertible illumination with orange light. Blue caused specific coupling Gi, sustained Gi dissociation, decreased intracellular cAMP levels, activation GIRK channels. responses activated were partially terminated illumination. These light-dependent indicate that InvC-opsin is a typical bistable pigment wherein resting can be interconverted visible We attempted to modulate functional using site-directed mutagenesis. Substitution Ser-94 Ala little shift state further red ∼10 nm state, indicating spectra tuned differently. contrast, substitution S94A did significantly affect AcrInvC-opsin. Because blue-sensitive, Gi/o-biased, pigment, it has potential serve as optical control tool specifically reversibly regulate Gi/o-dependent pathways

Язык: Английский

Процитировано

1

Dynorphin modulates reward-seeking actions through a pallido-amygdala cholinergic circuit DOI
Qingtao Sun, Mingzhe Liu, Wuqiang Guan

и другие.

Neuron, Год журнала: 2025, Номер unknown

Опубликована: Апрель 1, 2025

Язык: Английский

Процитировано

1