AI Chat Paper
Note: Please note that the following content is generated by AMiner AI. SciOpen does not take any responsibility related to this content.
{{lang === 'zh_CN' ? '文章概述' : 'Summary'}}
{{lang === 'en_US' ? '中' : 'Eng'}}
Chat more with AI
PDF (2.8 MB)
Collect
Submit Manuscript AI Chat Paper
Show Outline
Outline
Show full outline
Hide outline
Outline
Show full outline
Hide outline
Review | Open Access

Exploring lysosomal biology: current approaches and methods

State Key Laboratory of Conservation and Utilization of Bio-resources in Yunnan and Center for Life Sciences, School of Life Sciences, Yunnan University, Kunming 650091, China
Southwest United Graduate School, Kunming 650092, China
Show Author Information

Graphical Abstract

Abstract

Lysosomes are the degradation centers and signaling hubs in the cell. Lysosomes undergo adaptation to maintain cell homeostasis in response to a wide variety of cues. Dysfunction of lysosomes leads to aging and severe diseases including lysosomal storage diseases (LSDs), neurodegenerative disorders, and cancer. To understand the complexity of lysosome biology, many research approaches and tools have been developed to investigate lysosomal functions and regulatory mechanisms in diverse experimental systems. This review summarizes the current approaches and tools adopted for studying lysosomes, and aims to provide a methodological overview of lysosomal research and related fields.

References

 

Aits S, Kricker J, Liu B, Ellegaard AM, Hamalisto S, Tvingsholm S, Corcelle-Termeau E, Hogh S, Farkas T, Holm Jonassen A, Gromova I, Mortensen M, Jaattela M (2015) Sensitive detection of lysosomal membrane permeabilization by lysosomal galectin puncta assay. Autophagy 11(8): 1408−1424

 

Anderson RG, Orci L (1988) A view of acidic intracellular compartments. Journal of Cell Biology 106(3): 539−543

 

Bagshaw RD, Mahuran DJ, Callahan JW (2005) A proteomic analysis of lysosomal integral membrane proteins reveals the diverse composition of the organelle. Mol Cell Proteomics 4(2): 133−143

 

Boonacker E, van Noorden CJ (2001) Enzyme cytochemical techniques for metabolic mapping in living cells, with special reference to proteolysis. J Histochem Cytochem 49(12): 1473−1486

 

Bright NA, Davis LJ, Luzio JP (2016) Endolysosomes Are the principal intracellular sites of acid hydrolase activity. Curr Biol 26(17): 2233−2245

 

Carmona-Gutierrez D, Hughes AL, Madeo F, Ruckenstuhl C (2016) The crucial impact of lysosomes in aging and longevity. Ageing Res Rev 32: 2−12

 

Castro-Mondragon JA, Riudavets-Puig R, Rauluseviciute I, Lemma RB, Turchi L, Blanc-Mathieu R, Lucas J, Boddie P, Khan A, Manosalva Perez N, Fornes O, Leung TY, Aguirre A, Hammal F, Schmelter D, Baranasic D, Ballester B, Sandelin A, Lenhard B, Vandepoele K, Wasserman WW, Parcy F, Mathelier A (2022) JASPAR 2022: the 9th release of the open-access database of transcription factor binding profiles. Nucleic Acids Res 50(D1): D165−D173

 

Chen CS, Chen WN, Zhou M, Arttamangkul S, Haugland RP (2000) Probing the cathepsin D using a BODIPY FL-pepstatin A: applications in fluorescence polarization and microscopy. J Biochem Biophys Methods 42(3): 137−151

 

Chen D, Xiao H, Zhang K, Wang B, Gao Z, Jian Y, Qi X, Sun J, Miao L, Yang C (2010) Retromer is required for apoptotic cell clearance by phagocytic receptor recycling. Science 327(5970): 1261−1264

 

Cheng XT, Xie YX, Zhou B, Huang N, Farfel-Becker T, Sheng ZH (2018) Characterization of LAMP1-labeled nondegradative lysosomal and endocytic compartments in neurons. J Cell Biol 217(9): 3127−3139

 

Chin MY, Patwardhan AR, Ang KH, Wang AL, Alquezar C, Welch M, Nguyen PT, Grabe M, Molofsky AV, Arkin MR, Kao AW (2021) Genetically encoded, pH-sensitive mTFP1 biosensor for probing lysosomal pH. ACS Sens 6(6): 2168−2180

 

Cinque L, De Leonibus C, Iavazzo M, Krahmer N, Intartaglia D, Salierno FG, De Cegli R, Di Malta C, Svelto M, Lanzara C, Maddaluno M, Wanderlingh LG, Huebner AK, Cesana M, Bonn F, Polishchuk E, Hubner CA, Conte I, Dikic I, Mann M, Ballabio A, Sacco F, Grumati P, Settembre C (2020) MiT/TFE factors control ER-phagy via transcriptional regulation of FAM134B. EMBO J 39(17): e105696. https://doi.org/10.15252/embj.2020105696

 

Corrotte M, Castro-Gomes T (2019) Lysosomes and plasma membrane repair. Curr Top Membr 84: 1−16

 

Davidson SM, Vander Heiden MG (2017) Critical functions of the lysosome in cancer biology. Annu Rev Pharmacol Toxicol 57: 481−507

 

de Duve C (2005) The lysosome turns fifty. Nat Cell Biol 7(9): 847−849

 

DePedro HM, Urayama P (2009) Using LysoSensor Yellow/Blue DND-160 to sense acidic pH under high hydrostatic pressures. Anal Biochem 384(2): 359−361

 

Diwu Z, Chen CS, Zhang C, Klaubert DH, Haugland RP (1999) A novel acidotropic pH indicator and its potential application in labeling acidic organelles of live cells. Chem Biol 6(7): 411−418

 

Doronzo G, Astanina E, Cora D, Chiabotto G, Comunanza V, Noghero A, Neri F, Puliafito A, Primo L, Spampanato C, Settembre C, Ballabio A, Camussi G, Oliviero S, Bussolino F (2019) TFEB controls vascular development by regulating the proliferation of endothelial cells. EMBO J 38(3): e98250. https://doi.org/10.15252/embj.201798250

 

Elrick MJ, Pacheco CD, Yu T, Dadgar N, Shakkottai VG, Ware C, Paulson HL, Lieberman AP (2010) Conditional Niemann-Pick C mice demonstrate cell autonomous Purkinje cell neurodegeneration. Hum Mol Genet 19(5): 837−847

 

Eriksson I, Vainikka L, Persson HL, Ollinger K (2023) Real-time monitoring of lysosomal membrane permeabilization using acridine orange. Methods Protoc 6(4): 72. https://doi.org/10.3390/mps6040072

 
Falcon-Perez JM, Nazarian R, Sabatti C, Dell'Angelica EC (2005) Distribution and dynamics of Lamp1-containing endocytic organelles in fibroblasts deficient in BLOC-3. J Cell Sci 118(Pt 22): 5243−5255
 

Farias GG, Guardia CM, De Pace R, Britt DJ, Bonifacino JS (2017) BORC/kinesin-1 ensemble drives polarized transport of lysosomes into the axon. Proc Natl Acad Sci USA 114(14): E2955−E2964

 

Frost LS, Dhingra A, Reyes-Reveles J, Boesze-Battaglia K (2017) The use of DQ-BSA to monitor the turnover of autophagy-associated cargo. Methods Enzymol 587: 43−54

 
Gan QW, Wang X, Zhang Q, Yin QY, Jiang YL, Liu YB, Xuan N, Li JL, Zhou JX, Liu K, Jing YD, Wang XC, Yang C (2019) The amino acid transporter SLC-36.1 cooperates with PtdIns3P 5-kinase to control phagocytic lysosome reformation. J Cell Biol 218(8): 2619−2637
 

Gieselmann V, Hasilik A, von Figura K (1985) Processing of human cathepsin D in lysosomes in vitro. J Biol Chem 260(5): 3215−3220

 

Guo P, Hu T, Zhang J, Jiang S, Wang X (2010) Sequential action of Caenorhabditis elegans Rab GTPases regulates phagolysosome formation during apoptotic cell degradation. Proc Natl Acad Sci USA 107(42): 18016−18021

 

Hart PD, Young MR (1991) Ammonium chloride, an inhibitor of phagosome-lysosome fusion in macrophages, concurrently induces phagosome-endosome fusion, and opens a novel pathway: studies of a pathogenic mycobacterium and a nonpathogenic yeast. J Exp Med 174(4): 881−889

 

Hirst J, Futter CE, Hopkins CR (1998) The kinetics of mannose 6-phosphate receptor trafficking in the endocytic pathway in HEp-2 cells: the receptor enters and rapidly leaves multivesicular endosomes without accumulating in a prelysosomal compartment. Mol Biol Cell 9(4): 809−816

 

Homewood CA, Warhurst DC, Peters W, Baggaley VC (1972) Lysosomes, pH and the anti-malarial action of chloroquine. Nature 235(5332): 50−52

 
Hu M, Li P, Wang C, Feng X, Geng Q, Chen W, Marthi M, Zhang W, Gao C, Reid W, Swanson J, Du W, Hume RI, Xu H (2022) Parkinson's disease-risk protein TMEM175 is a proton-activated proton channel in lysosomes. Cell 185(13): 2292−2308
 

Huss M, Ingenhorst G, Konig S, Gassel M, Drose S, Zeeck A, Altendorf K, Wieczorek H (2002) Concanamycin A, the specific inhibitor of V-ATPases, binds to the V(o) subunit c. J Biol Chem 277(43): 40544−40548

 
Jia J, Claude-Taupin A, Gu Y, Choi SW, Peters R, Bissa B, Mudd MH, Allers L, Pallikkuth S, Lidke KA, Salemi M, Phinney B, Mari M, Reggiori F, Deretic V (2020) Galectin-3 coordinates a cellular system for lysosomal repair and removal. Dev Cell 52(1): 69−87
 

Khaminets A, Heinrich T, Mari M, Grumati P, Huebner AK, Akutsu M, Liebmann L, Stolz A, Nietzsche S, Koch N, Mauthe M, Katona I, Qualmann B, Weis J, Reggiori F, Kurth I, Hubner CA, Dikic I (2015) Regulation of endoplasmic reticulum turnover by selective autophagy. Nature 522(7556): 354−358

 

Kimura S, Noda T, Yoshimori T (2007) Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 3(5): 452−460

 

Kobayashi T, Vischer UM, Rosnoblet C, Lebrand C, Lindsay M, Parton RG, Kruithof EK, Gruenberg J (2000) The tetraspanin CD63/lamp3 cycles between endocytic and secretory compartments in human endothelial cells. Mol Biol Cell 11(5): 1829−1843

 

Kuroda T, Yoshinari M, Okamura K, Okazawa K, Ikenoue H, Sato K, Fujishima M (1994) Effects of lysosomal protease inhibitors on the degradation of acetylated low density lipoprotein in cultured rat peritoneal macrophages. J Atheroscler Thromb 1(1): 41−44

 

Laqtom NN, Dong W, Medoh UN, Cangelosi AL, Dharamdasani V, Chan SH, Kunchok T, Lewis CA, Heinze I, Tang R, Grimm C, Dang Do AN, Porter FD, Ori A, Sabatini DM, Abu-Remaileh M (2022) CLN3 is required for the clearance of glycerophosphodiesters from lysosomes. Nature 609(7929): 1005−1011

 

Lee JH, Yang DS, Goulbourne CN, Im E, Stavrides P, Pensalfini A, Chan H, Bouchet-Marquis C, Bleiwas C, Berg MJ, Huo C, Peddy J, Pawlik M, Levy E, Rao M, Staufenbiel M, Nixon RA (2022) Faulty autolysosome acidification in Alzheimer's disease mouse models induces autophagic build-up of Abeta in neurons, yielding senile plaques. Nat Neurosci 25(6): 688−701

 
Lencer WI, Weyer P, Verkman AS, Ausiello DA, Brown D (1990) FITC-dextran as a probe for endosome function and localization in kidney. Am J Physiol 258(2 Pt 1): C309−317
 

Li Y, Chen B, Zou W, Wang X, Wu Y, Zhao D, Sun Y, Liu Y, Chen L, Miao L, Yang C, Wang X (2016a) The lysosomal membrane protein SCAV-3 maintains lysosome integrity and adult longevity. J Cell Biol 215(2): 167−185

 

Li Y, Xu M, Ding X, Yan C, Song Z, Chen L, Huang X, Wang X, Jian Y, Tang G, Tang C, Di Y, Mu S, Liu X, Liu K, Li T, Wang Y, Miao L, Guo W, Hao X, Yang C (2016b) Protein kinase C controls lysosome biogenesis independently of mTORC1. Nat Cell Biol 18(10): 1065−1077

 

Lin HJ, Herman P, Kang JS, Lakowicz JR (2001) Fluorescence lifetime characterization of novel low-pH probes. Anal Biochem 294(2): 118−125

 

Liou B, Zhang W, Fannin V, Quinn B, Ran H, Xu K, Setchell KDR, Witte D, Grabowski GA, Sun Y (2019) Combination of acid beta-glucosidase mutation and Saposin C deficiency in mice reveals Gba1 mutation dependent and tissue-specific disease phenotype. Sci Rep 9(1): 5571. https://doi.org/10.1038/s41598-019-41914-7

 

Liu B, Du H, Rutkowski R, Gartner A, Wang X (2012) LAAT-1 is the lysosomal lysine/arginine transporter that maintains amino acid homeostasis. Science 337(6092): 351−354

 

Liu K, Jian Y, Sun X, Yang C, Gao Z, Zhang Z, Liu X, Li Y, Xu J, Jing Y, Mitani S, He S, Yang C (2016) Negative regulation of phosphatidylinositol 3-phosphate levels in early-to-late endosome conversion. J Cell Biol 212(2): 181−198

 

Lloyd-Evans E, Waller-Evans H (2020) Lysosomal Ca2 + homeostasis and signaling in health and disease. Cold Spring Harb Perspect Biol 12(6): a035311. https://doi.org/10.1101/cshperspect.a035311

 

Luzio JP, Pryor PR, Bright NA (2007) Lysosomes: fusion and function. Nat Rev Mol Cell Biol 8(8): 622−632

 

Maejima I, Takahashi A, Omori H, Kimura T, Takabatake Y, Saitoh T, Yamamoto A, Hamasaki M, Noda T, Isaka Y, Yoshimori T (2013) Autophagy sequesters damaged lysosomes to control lysosomal biogenesis and kidney injury. EMBO J 32(17): 2336−2347

 

Mansueto G, Armani A, Viscomi C, D'Orsi L, De Cegli R, Polishchuk EV, Lamperti C, Di Meo I, Romanello V, Marchet S, Saha PK, Zong H, Blaauw B, Solagna F, Tezze C, Grumati P, Bonaldo P, Pessin JE, Zeviani M, Sandri M, Ballabio A (2017) Transcription factor EB controls metabolic flexibility during exercise. Cell Metab 25(1): 182−196

 

Marwaha R, Sharma M (2017) DQ-Red BSA trafficking assay in cultured cells to assess cargo delivery to lysosomes. Bio Protoc 7(19): e2571. https://doi.org/10.21769/BioProtoc.2571

 

Meng Y, Heybrock S, Neculai D, Saftig P (2020) Cholesterol handling in lysosomes and beyond. Trends Cell Biol 30(6): 452−466

 
Miao R, Li M, Zhang Q, Yang C, Wang X (2020) An ECM-to-nucleus signaling pathway activates lysosomes for C. elegans larval development. Dev Cell 52(1): 21−37
 

Mindell JA (2012) Lysosomal acidification mechanisms. Annu Rev Physiol 74: 69−86

 

Mizushima N, Yamamoto A, Matsui M, Yoshimori T, Ohsumi Y (2004) In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol Biol Cell 15(3): 1101−1111

 

Muthukottiappan P, Winter D (2021) A proteomic view on lysosomes. Mol Omics 17(6): 842−859

 

Oben JA, Foreman JC (1988) A simple quantitative fluorimetric assay of in vitro phagocytosis in human neutrophils. J Immunol Methods 112(1): 99−103

 

Perera RM, Zoncu R (2016) The Lysosome as a Regulatory Hub. Annu Rev Cell Dev Biol 32: 223−253

 

Pickles S, Vigie P, Youle RJ (2018) Mitophagy and quality control mechanisms in mitochondrial maintenance. Curr Biol 28(4): R170−R185

 

Platt FM, d'Azzo A, Davidson BL, Neufeld EF, Tifft CJ (2018) Lysosomal storage diseases. Nat Rev Dis Primers 4(1): 27. https://doi.org/10.1038/s41572-018-0025-4

 

Ponsford AH, Ryan TA, Raimondi A, Cocucci E, Wycislo SA, Frohlich F, Swan LE, Stagi M (2021) Live imaging of intra-lysosome pH in cell lines and primary neuronal culture using a novel genetically encoded biosensor. Autophagy 17(6): 1500−1518

 

Puertollano R, Ferguson SM, Brugarolas J, Ballabio A (2018) The complex relationship between TFEB transcription factor phosphorylation and subcellular localization. EMBO J 37(11): e98804. https://doi.org/10.15252/embj.201798804

 

Raben N, Nagaraju K, Lee E, Kessler P, Byrne B, Lee L, LaMarca M, King C, Ward J, Sauer B, Plotz P (1998) Targeted disruption of the acid alpha-glucosidase gene in mice causes an illness with critical features of both infantile and adult human glycogen storage disease type II. J Biol Chem 273(30): 19086−19092

 

Raben N, Puertollano R (2016) TFEB and TFE3: linking lysosomes to cellular adaptation to stress. Annu Rev Cell Dev Biol 32: 255−278

 

Roczniak-Ferguson A, Petit CS, Froehlich F, Qian S, Ky J, Angarola B, Walther TC, Ferguson SM (2012) The transcription factor TFEB links mTORC1 signaling to transcriptional control of lysosome homeostasis. Sci Signal 5(228): ra42. https://doi.org/10.1126/scisignal.2002790

 

Saftig P, Klumperman J (2009) Lysosome biogenesis and lysosomal membrane proteins: trafficking meets function. Nat Rev Mol Cell Biol 10(9): 623−635

 

Sardiello M, Palmieri M, di Ronza A, Medina DL, Valenza M, Gennarino VA, Di Malta C, Donaudy F, Embrione V, Polishchuk RS, Banfi S, Parenti G, Cattaneo E, Ballabio A (2009) A gene network regulating lysosomal biogenesis and function. Science 325(5939): 473−477

 

Schroder B, Wrocklage C, Hasilik A, Saftig P (2010) Molecular characterisation of 'transmembrane protein 192' (TMEM192), a novel protein of the lysosomal membrane. Biol Chem 391(6): 695−704

 

Settembre C, De Cegli R, Mansueto G, Saha PK, Vetrini F, Visvikis O, Huynh T, Carissimo A, Palmer D, Klisch TJ, Wollenberg AC, Di Bernardo D, Chan L, Irazoqui JE, Ballabio A (2013) TFEB controls cellular lipid metabolism through a starvation-induced autoregulatory loop. Nat Cell Biol 15(6): 647−658

 

Settembre C, Di Malta C, Polito VA, Garcia-Arencibia M, Vetrini F, Erdin S, Erdin SU, Huynh T, Medina D, Colella P, Sardiello M, Rubinsztein DC, Ballabio A (2011) TFEB links autophagy to lysosomal biogenesis. Science 332(6036): 1429−1433

 

Steingrimsson E, Tessarollo L, Reid SW, Jenkins NA, Copeland NG (1998) The bHLH-Zip transcription factor Tfeb is essential for placental vascularization. Development 125(23): 4607−4616

 

Sun Y, Li M, Zhao D, Li X, Yang C, Wang X (2020) Lysosome activity is modulated by multiple longevity pathways and is important for lifespan extension in C. elegans. Elife 9: e55745. https://doi.org/10.7554/eLife.55745

 

Tancini B, Buratta S, Delo F, Sagini K, Chiaradia E, Pellegrino RM, Emiliani C, Urbanelli L (2020) Lysosomal exocytosis: the extracellular role of an intracellular organelle. Membranes (Basel) 10(12): 406. https://doi.org/10.3390/membranes10120406

 

Thelen AM, Zoncu R (2017) Emerging roles for the lysosome in lipid metabolism. Trends Cell Biol 27(11): 833−850

 

Titus EO (1989) Recent developments in the understanding of the pharmacokinetics and mechanism of action of chloroquine. Ther Drug Monit 11(4): 369−379

 

Turk V, Stoka V, Vasiljeva O, Renko M, Sun T, Turk B, Turk D (2012) Cysteine cathepsins: from structure, function and regulation to new frontiers. Biochim Biophys Acta 1824(1): 68−88

 

Udayar V, Chen Y, Sidransky E, Jagasia R (2022) Lysosomal dysfunction in neurodegeneration: emerging concepts and methods. Trends Neurosci 45(3): 184−199

 

Van Noorden CJ, Boonacker E, Bissell ER, Meijer AJ, van Marle J, Smith RE (1997) Ala-Pro-cresyl violet, a synthetic fluorogenic substrate for the analysis of kinetic parameters of dipeptidyl peptidase IV (CD26) in individual living rat hepatocytes. Anal Biochem 252(1): 71−77

 

Wang F, Gomez-Sintes R, Boya P (2018) Lysosomal membrane permeabilization and cell death. Traffic 19(12): 918−931

 

Wang Z, Zhao H, Yuan C, Zhao D, Sun Y, Wang X, Zhang H (2019) The RBG-1-RBG-2 complex modulates autophagy activity by regulating lysosomal biogenesis and function in C. elegans. J Cell Sci 132(19): jcs234195. https://doi.org/10.1242/jcs.234195

 

Webb BA, Aloisio FM, Charafeddine RA, Cook J, Wittmann T, Barber DL (2021) pHLARE: a new biosensor reveals decreased lysosome pH in cancer cells. Mol Biol Cell 32(2): 131−142

 

Xie YX, Naseri NN, Fels J, Kharel P, Na Y, Lane D, Burre J, Sharma M (2022) Lysosomal exocytosis releases pathogenic alpha-synuclein species from neurons in synucleinopathy models. Nat Commun 13(1): 4918. https://doi.org/10.1038/s41467-022-32625-1

 

Xing R, Zhou H, Jian Y, Li L, Wang M, Liu N, Yin Q, Liang Z, Guo W, Yang C (2021) The Rab7 effector WDR91 promotes autophagy-lysosome degradation in neurons by regulating lysosome fusion. J Cell Biol 220(8): e202007061. https://doi.org/10.1083/jcb.202007061

 

Xu M, Liu Y, Zhao L, Gan Q, Wang X, Yang C (2014) The lysosomal cathepsin protease CPL-1 plays a leading role in phagosomal degradation of apoptotic cells in Caenorhabditis elegans. Mol Biol Cell 25(13): 2071−2083

 

Yadati T, Houben T, Bitorina A, Shiri-Sverdlov R (2020) The ins and outs of cathepsins: physiological function and role in disease management. Cells 9(7): 1679. https://doi.org/10.3390/cells9071679

 

Yang C, Wang X (2017) Cell biology in China: focusing on the lysosome. Traffic 18(6): 348−357

 

Yang C, Wang X (2021) Lysosome biogenesis: regulation and functions. J Cell Biol 220(6): e202102001. https://doi.org/10.1083/jcb.202102001

 

Yin Q, Jian Y, Xu M, Huang X, Wang N, Liu Z, Li Q, Li J, Zhou H, Xu L, Wang Y, Yang C (2020) CDK4/6 regulate lysosome biogenesis through TFEB/TFE3. J Cell Biol 219(8): e201911036. https://doi.org/10.1083/jcb.201911036

 

Yoshimori T, Yamamoto A, Moriyama Y, Futai M, Tashiro Y (1991) Bafilomycin A1, a specific inhibitor of vacuolar-type H( + )-ATPase, inhibits acidification and protein degradation in lysosomes of cultured cells. J Biol Chem 266(26): 17707−17712

Biophysics Reports
Pages 111-120
Cite this article:
Yin Q, Yang C. Exploring lysosomal biology: current approaches and methods. Biophysics Reports, 2024, 10(2): 111-120. https://doi.org/10.52601/bpr.2023.230028

132

Views

4

Downloads

0

Crossref

0

Scopus

0

CSCD

Altmetrics

Received: 28 October 2023
Accepted: 04 January 2024
Published: 30 April 2024
© The Author(s) 2024

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Return