Abstract
Neuronal circuits in the neocortex and hippocampus are essential for higher brain functions such as motor learning and spatial memory. In the mammalian forebrain, most excitatory synapses of pyramidal neurons are formed on spines, which are tiny protrusions extending from the dendritic shaft. The spine contains specialized molecular machinery that regulates synaptic transmission and plasticity. Spine size correlates with the efficacy of synaptic transmission, and spine morphology affects signal transduction at the post-synaptic compartment. Plasticity-related changes in the structural and molecular organization of spine synapses are thought to underlie the cellular basis of learning and memory. Recent advances in super-resolution microscopy have revealed the molecular mechanisms of the nanoscale synaptic structures regulating synaptic transmission and plasticity in living neurons, which are difficult to investigate using electron microscopy alone. In this review, we summarize recent advances in super-resolution imaging of spine synapses and discuss the implications of nanoscale structures in the regulation of synaptic function, learning, and memory.
Similar content being viewed by others
References
Arellano JI (2007) Ultrastructure of dendritic spines: correlation between synaptic and spine morphologies. Front Neurosci 1:131–143
Attardo A, Fitzgerald JE, Schnitzer MJ (2015) Impermanence of dendritic spines in live adult CA1 hippocampus. Nature 523:592–596
Baddeley D, Bewersdorf J (2018) Biological insight from super-resolution microscopy: what we can learn from localization-based images. Annu Rev Biochem 87:965–989
Bär J, Kobler O, Van Bommel B, Mikhaylova M (2016) Periodic F-actin structures shape the neck of dendritic spines. Sci Rep 6:1–9
Berning S, Willig KI, Steffens H et al (2012) Nanoscopy in a living mouse brain. Science 335:551
Bethge P, Chéreau R, Avignone E et al (2013) Two-photon excitation STED microscopy in two colors in acute brain slices. Biophys J 104:778–785
Betzig E, Patterson GH, Sougrat R et al (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642–1645
Biederer T, Kaeser PS, Blanpied TA (2017) Transcellular nanoalignment of synaptic function. Neuron 96:680–696
Bosch M, Castro J, Saneyoshi T et al (2014) Structural and molecular remodeling of dendritic spine substructures during long-term potentiation. Neuron 82:444–459
Bourne JN, Harris KM (2008) Balancing structure and function at hippocampal dendritic spines. Annu Rev Neurosci 31:47–67
Brakemann T, Stiel AC, Weber G et al (2011) A reversibly photoswitchable GFP-like protein with fluorescence excitation decoupled from switching. Nat Biotechnol 29:942–950
Broadhead MJ, Horrocks MH, Zhu F et al (2016) PSD95 nano-clusters are post-synaptic building blocks in hippocampus circuits. Sci Rep 6:1–14
Chamma I, Letellier M, Butler C et al (2016) Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin. Nat Commun 7:1–15
Chang JB, Chen F, Yoon YG et al (2017) Iterative expansion microscopy. Nat Methods 14:593–599
Chazeau A, Mehidi A, Nair D et al (2014) Nanoscale segregation of actin nucleation and elongation factors determines dendritic spine protrusion. EMBO J. https://doi.org/10.15252/embj.201488837
Chen F, Tillberg PW, Boyden ES (2015) Expansion microscopy. Science 347:543–548
Chen F, Wassie AT, Cote AJ et al (2016) Nanoscale imaging of RNA with expansion microscopy. Nat Methods 13:679–684
Chen H, Tang AH, Blanpied TA (2018) Subsynaptic spatial organization as a regulator of synaptic strength and plasticity. Curr Opin Neurobiol 51:147–153
Choquet D (2018) Linking nanoscale dynamics of AMPA receptor organization to plasticity of excitatory synapses and learning. J Neurosci 38:9318–9329
Chéreau R, Tønnesen J, Nägerl UV (2015) STED microscopy for nanoscale imaging in living brain slices. Methods 88:57–66
Cho T, Kashiwagi Y, Okabe S (2019) Temporal sequences of synapse disintegration triggered by afferent axon transection, time-lapse imaging study of pre-synaptic and post-synaptic molecules. eNeuro 6:1–13
Chozinski TJ, Halpern AR, Okawa H et al (2016) Expansion microscopy with conventional antibodies and fluorescent proteins. Nat Methods 13:485–488
D’Este E, Kamin D, Göttfert F et al (2015) STED nanoscopy reveals the ubiquity of subcortical cytoskeleton periodicity in living neurons. Cell Rep 10:1246–1251
Dani A, Huang B, Bergan J et al (2010) Superresolution imaging of chemical synapses in the brain. Neuron 68:843–856
Danzl JG, Sidenstein SC, Gregor C et al (2016) Coordinate-targeted fluorescence nanoscopy with multiple off states. Nat Photonics 10:122–128
De Luca GMR, Breedijk RMP, Brandt RAJ et al (2013) Re-scan confocal microscopy: scanning twice for better resolution. Biomed Opt Express 4:2644
Diering GH, Huganir RL (2018) The AMPA receptor code of synaptic plasticity. Neuron 100:314–329
Fifková E, Anderson CL (1981) Stimulation-induced changes in dimensions of stalks of dendritic spines in the dentate molecular layer. Exp Neurol 74:621–627
Frost NA, Shroff H, Kong H et al (2010) Single-molecule discrimination of discrete perisynaptic and distributed sites of actin filament assembly within dendritic spines. Neuron 67:86–99
Fukata Y, Dimitrov A, Boncompain G et al (2013) Local palmitoylation cycles define activity-regulated post-synaptic subdomains. J Cell Biol 202:145–161
Gao R, Asano S, Upadhyayula S et al (2019) Cortical column and whole-brain imaging with molecular contrast and nanoscale resolution. Science 363:eaau8302
Giannone G, Hosy E, Levet F et al (2010) Dynamic super-resolution imaging of endogenous proteins on living cells at ultra-high density. Biophys J 99:1303–1310
Glebov OO, Cox S, Humphreys L, Burrone J (2016) Neuronal activity controls transsynaptic geometry. Sci Rep. https://doi.org/10.1038/srep22703
Glebov OO, Jackson RE, Winterflood CM et al (2017) Nanoscale structural plasticity of the active zone matrix modulates pre-synaptic function. Cell Rep 18:2715–2728
Göttfert F, Pleiner T, Heine J et al (2017) Strong signal increase in STED fluorescence microscopy by imaging regions of subdiffraction extent. Proc Natl Acad Sci USA 114:2125–2130
Griffiths VA, Valera AM, Lau JYN et al (2020) Real-time 3D movement correction for two-photon imaging in behaving animals. Nat Methods 17:741–748
Grotjohann T, Testa I, Leutenegger M et al (2011) Diffraction-unlimited all-optical imaging and writing with a photochromic GFP. Nature 478:204–208
Guo Y, Di LD, Zhang S et al (2018) Visualizing intracellular organelle and cytoskeletal interactions at nanoscale resolution on millisecond timescales. Cell 175:1–13
Gustafsson MGL (2000) Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J Microsc 198:82–87
Gustafsson MGL (2005) Nonlinear structured-illumination microscopy: Widefield fluorescence imaging with theoretically unlimited resolution. Proc Natl Acad Sci USA 102:13081–13086
Gustafsson MGLL, Shao L, Carlton PM et al (2008) Three-dimensional resolution doubling in widefield fluorescence microscopy by structured illumination. Biophys J 94:4957–4970
Gwosch KC, Pape JK, Balzarotti F et al (2020) MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells. Nat Methods 17:217–224
Harris KM (2020) Synaptic odyssey. J Neurosci 40:61–80
Harris KM, Jensen FE, Tsao B (1992) Three-dimensional structure of dendritic spines and synapses in rat hippocampus (CA1) at postnatal day 15 and adult ages: implications for the maturation of synaptic physiology and long-term potentiation. J Neurosci 12:2685–2705
Hayashi S, Okada Y (2015) Ultrafast super-resolution fluorescence imaging with spinning disk confocal microscope optics. Mol Biol Cell 26:1743–1751
He J, Zhou R, Wu Z et al (2016) Prevalent presence of periodic actin-spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species. Proc Natl Acad Sci USA 113:6029–6034
Heine J, Reuss M, Harke B et al (2017) Adaptive-illumination STED nanoscopy. Proc Natl Acad Sci USA 114:9797–9802
Heintzmann R, Cremer CG (1999) Laterally modulated excitation microscopy: improvement of resolution by using a diffraction grating. Proc SPIE 3568 Opt Biopsies Microsc Tech III. https://doi.org/10.1117/12.336833
Heintzmann R, Huser T (2017) Super-resolution structured illumination microscopy. Chem Rev 117:13890–13908
Hell SW (2003) Toward fluorescence nanoscopy. Nat Biotechnol 21:1347–1355
Hell SW, Wichmann J (1994) Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt Lett 19:780
Helmchen F, Denk W (2005) Deep tissue two-photon microscopy. Nat Methods 2:932–940
Hering H, Sheng M (2001) Dendritic spines: structure, dynamics and regulation. Nat Rev Neurosci 2:880–888
Hess ST, Girirajan TPK, Mason MD (2006) Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. Biophys J 91:4258–4272
Higashi T, Tanaka S, Ida T, Okabe S (2018) Synapse elimination triggered by BMP4 exocytosis and pre-synaptic BMP receptor activation. Cell Rep 22:919–929
Hirano Y, Matsuda A, Hiraoka Y (2015) Recent advancements in structured-illumination microscopy toward live-cell imaging. Microscopy 64:237–249
Hirokawa N (1989) The arrangement of actin filaments in the post-synaptic cytoplasm of the cerebellar cortex revealed by quick-freeze deep-etch electron microscopy. Neurosci Res 6:269–275
Hofmann M, Eggeling C, Jakobs S, Hell SW (2005) Breaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteins. Proc Natl Acad Sci USA 102:17565–17569
Holtmaat A, Svoboda K (2009) Experience-dependent structural synaptic plasticity in the mammalian brain. Nat Rev Neurosci 10:647–658
Honkura N, Matsuzaki M, Noguchi J et al (2008) The subspine organization of actin fibers regulates the structure and plasticity of dendritic spines. Neuron 57:719–729
Hotulainen P, Hoogenraad CC (2010) Actin in dendritic spines: connecting dynamics to function. J Cell Biol 189:619–629
Hruska M, Henderson N, Le Marchand SJ et al (2018) Synaptic nanomodules underlie the organization and plasticity of spine synapses. Nat Neurosci 21:671–682
Huang B, Jones SA, Brandenburg B, Zhuang X (2008) Whole-cell 3D STORM reveals interactions between cellular structures with nanometer-scale resolution. Nat Methods 5:1047–1052
Inavalli VVGK, Lenz MO, Butler C et al (2019) A super-resolution platform for correlative live single-molecule imaging and STED microscopy. Nat Methods 16:1263–1268
Isshiki M, Okabe S (2014) Evaluation of cranial window types for in vivo two-photon imaging of brain microstructures. Microscopy 63:53–63
Isshiki M, Tanaka S, Kuriu T et al (2014) Enhanced synapse remodelling as a common phenotype in mouse models of autism. Nat Commun 5:1–15
Iwasaki K, Obashi K, Okabe S (2020) Vasodilator-stimulated phosphoprotein (VASP) is recruited into dendritic spines via G-actin-dependent mechanism and contributes to spine enlargement and stabilization. Eur J Neurosci 51:806–821
Izeddin I, Specht CG, Lelek M et al (2011) Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe. PLoS ONE 6:e15611
Ji N (2017) Adaptive optical fluorescence microscopy. Nat Methods 14:374–380
Kasai H, Fukuda M, Watanabe S et al (2010) Structural dynamics of dendritic spines in memory and cognition. Trends Neurosci 33:121–129
Kashiwagi Y, Higashi T, Obashi K et al (2019) Computational geometry analysis of dendritic spines by structured illumination microscopy. Nat Commun 10:1–14
Kawabata I, Kashiwagi Y, Obashi K et al (2012) LIS1-dependent retrograde translocation of excitatory synapses in developing interneuron dendrites. Nat Commun 3:722
Ke MT, Nakai Y, Fujimoto S et al (2016) Super-resolution mapping of neuronal circuitry with an index-optimized clearing agent. Cell Rep 14:2718–2732
Kim E, Sheng M (2004) PDZ domain proteins of synapses. Nat Rev Neurosci 5:771–781
Klar TA, Jakobs S, Dyba M et al (2000) Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission. Proc Natl Acad Sci USA 97:8206–8210
Knott GW, Holtmaat A, Wilbrecht L et al (2006) Spine growth precedes synapse formation in the adult neocortex in vivo. Nat Neurosci 9:1117–1124
Korobova F, Svitkina T (2010) Molecular architecture of synaptic actin cytoskeleton in hippocampal neurons reveals a mechanism of dendritic spine morphogenesis. Mol Biol Cell 21:165–176
Kuriu T, Inoue A, Bito H et al (2006) Differential control of post-synaptic density scaffolds via actin-dependent and -independent mechanisms. J Neurosci 26:7693–7706
Leterrier C, Dubey P, Roy S (2017) The nano-architecture of the axonal cytoskeleton. Nat Rev Neurosci 18:713–726
Li D, Shao L, Chen BC et al (2015) Extended-resolution structured illumination imaging of endocytic and cytoskeletal dynamics. Science 349:aab3500
Li J, Sekine-Aizawa Y, Ebrahimi S et al (2019) Tumor suppressor protein CYLD regulates morphogenesis of dendrites and spines. Eur J Neurosci 50:2722–2739
Li Z, Zhang Q, Chou S-W et al (2020) Fast widefield imaging of neuronal structure and function with optical sectioning in vivo. Sci Adv 6:eaaz3870
Lukinavičius G, Reymond L, D’Este E et al (2014) Fluorogenic probes for live-cell imaging of the cytoskeleton. Nat Methods 11:731–733
MacGillavry HD, Song Y, Raghavachari S, Blanpied TA (2013) Nanoscale scaffolding domains within the post-synaptic density concentrate synaptic ampa receptors. Neuron 78:615–622
Makino H, Malinow R (2009) AMPA receptor incorporation into synapses during LTP: the role of lateral movement and exocytosis. Neuron 64:381–390
Mandracchia B, Hua X, Guo C et al (2020) Fast and accurate sCMOS noise correction for fluorescence microscopy. Nat Commun 11:1–12
Manley S, Gillette JM, Patterson GH et al (2008) High-density mapping of single-molecule trajectories with photoactivated localization microscopy. Nat Methods 5:155–157
Mao C, Lee M, Jhan J et al (2020) Feature-rich covalent stains for super-resolution and cleared tissue fluorescence microscopy. Sci Adv 6:eaba4542
Masch JM, Steffens H, Fischer J et al (2018) Robust nanoscopy of a synaptic protein in living mice by organic-fluorophore labeling. Proc Natl Acad Sci USA 115:E8047–E8056
Matsuzaki M, Ellis-Davies GCR, Nemoto T et al (2001) Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons. Nat Neurosci 4:1086–1092
Matsuzaki M, Honkura N, Ellis-Davies GCR, Kasai H (2004) Structural basis of long-term potentiation in single dendritic spines. Nature 429:761–766
Meyer D, Bonhoeffer T, Scheuss V (2014) Balance and stability of synaptic structures during synaptic plasticity. Neuron 82:430–443
Nägerl UV, Willig KI, Hein B et al (2008) Live-cell imaging of dendritic spines by STED microscopy. Proc Natl Acad Sci 105:18982–18987
Nair D, Hosy E, Petersen JD et al (2013) Super-resolution imaging reveals that AMPA receptors inside synapses are dynamically organized in nanodomains regulated by PSD95. J Neurosci 33:13204–13224
Nixon-Abell J, Obara CJ, Weigel AV et al (2016) Increased spatiotemporal resolution reveals highly dynamic dense tubular matrices in the peripheral ER. Science 354:3928
Obashi K, Okabe S (2013) Regulation of mitochondrial dynamics and distribution by synapse position and neuronal activity in the axon. Eur J Neurosci 38:2350–2363
Obashi K, Matsuda A, Inoue Y, Okabe S (2019) Precise remporal regulation of molecular diffusion within dendritic spines by actin polymers during structural plasticity. Cell Rep 27:1503-1515.e8
Okabe S (2007) Molecular anatomy of the post-synaptic density. Mol Cell Neurosci 34:503–518
Okabe S (2013) Fluorescence imaging of synapse formation and remodeling. Microscopy 62:51–62
Okabe S (2020a) Recent advances in computational methods for measurement of dendritic spines imaged by light microscopy. Microscopy 69:196–213
Okabe S (2020b) Regulation of actin dynamics in dendritic spines: Nanostructure, moelecular mobility, and signaling mechanisms. Mol Cell Neurosci. https://doi.org/10.1016/j.mcn.2020.103564
Okabe S, Kim HD, Miwa A et al (1999) Continual remodeling of post-synaptic density and its regulation by synaptic activity. Nat Neurosci 2:804–811
Okabe S, Urushido T, Konno D et al (2001) Rapid redistribution of the post-synaptic density protein PSD-Zip45 (Homer 1c) and Its differential regulation by NMDA receptors and calcium channels. J Neurosci 21:9561–9571
Patton BR, Burke D, Owald D et al (2016) Three-dimensional STED microscopy of aberrating tissue using dual adaptive optics. Opt Express 24:8862
Pfeiffer T, Poll S, Bancelin S et al (2018) Chronic 2P-STED imaging reveals high turnover of dendritic spines in the hippocampus in vivo. eLife. https://doi.org/10.7554/eLife.34700
Popov VI, Davies HA, Rogachevsky VV et al (2004) Remodelling of synaptic morphology but unchanged synaptic density during late phase long-term potentiation (LTP): a serial section electron micrograph study in the dentate gyrus in the anaesthetised rat. Neuroscience 128:251–262
Rego EH, Shao L, Macklin JJ et al (2012) Nonlinear structured-illumination microscopy with a photoswitchable protein reveals cellular structures at 50-nm resolution. Proc Natl Acad Sci USA 109:E135–E143
Richardson DS, Lichtman JW (2015) Clarifying tissue clearing. Cell 162:246–257
Rust MJ, Bates M, Zhuang X (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat Methods 3:793–795
Sahl SJ, Hell SW, Jakobs S (2017) Fluorescence nanoscopy in cell biology. Nat Rev Mol Cell Biol 18:685–701
Sakamoto H, Ariyoshi T, Kimpara N et al (2017) Synaptic weight set by Munc13-1 supramolecular assemblies. Nat Neurosci 21:41–49
Sawada K, Kawakami R, Shigemoto R, Nemoto T (2018) Super-resolution structural analysis of dendritic spines using three-dimensional structured illumination microscopy in cleared mouse brain slices. Eur J Neurosci 47:1033–1042
Schermelleh L, Heintzmann R, Leonhardt H (2010) A guide to super-resolution fluorescence microscopy. J Cell Biol 190:165–175
Schermelleh L, Ferrand A, Huser T et al (2019) Superresolution microscopy demystified. Nat Cell Biol 21:72–84
Schulz O, Pieper C, Clever M et al (2013) Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy. Proc Natl Acad Sci USA 110:21000–21005
Sheng M, Hoogenraad CC (2007) The post-synaptic architecture of excitatory synapses: a more quantitative view. Annu Rev Biochem 76:823–847
Shim SH, Xia C, Zhong G et al (2012) Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes. Proc Natl Acad Sci USA 109:13978–13983
Shin SM, Zhang N, Hansen J et al (2012) GKAP orchestrates activity-dependent post-synaptic protein remodeling and homeostatic scaling. Nat Neurosci 15:1655–1666
Siddig S, Aufmkolk S, Doose S et al (2020) Super-resolution imaging reveals the nanoscale organization of metabotropic glutamate receptors at pre-synaptic active zones. Sci Adv 6:eaay7193
Sidenstein SC, D’Este E, Böhm MJ et al (2016) Multicolour multilevel STED nanoscopy of actin/spectrin organization at synapses. Sci Rep 6:1–8
Sigal YMM, Speer CMM, Babcock HPP, Zhuang X (2015) Mapping synaptic input fields of neurons with super-resolution imaging. Cell 163:493–505
Smith KR, Kopeikina KJ, Fawcett-Patel JM et al (2014) Psychiatric risk factor ANK3/ankyrin-G nanodomains regulate the structure and function of glutamatergic synapses. Neuron 84:399–415
Sorra KE, Harris KM (2000) Overview on the structure, composition, function, development, and plasticity of hippocampal dendritic spines. Hippocampus 10:501–511
Spacek J, Harris KM (1997) Three-dimensional organization of smooth endoplasmic reticulum in hippocampal CA1 dendrites and dendritic spines of the immature and mature rat. J Neurosci 17:190–203
Spacek J, Harris KM (2004) Trans-endocytosis via spinules in adult rat hippocampus. J Neurosci 24:4233–4241
Spruston N (2008) Pyramidal neurons: dendritic structure and synaptic integration. Nat Rev Neurosci 9:206–221
Sugiyama Y, Kawabata I, Sobue K, Okabe S (2005) Determination of absolute protein numbers in single synapses by a GFP-based calibration technique. Nat Methods 2:677–684
Sydor AM, Czymmek KJ, Puchner EM, Mennella V (2015) Super-resolution microscopy: from single molecules to supramolecular assemblies. Trends Cell Biol 25:730–748
Takasaki KT, Ding JB, Sabatini BL (2013) Live-cell super-resolution imaging by pulsed STED two-photon excitation microscopy. Biophys J 104:770–777
Tang AH, Chen H, Li TP et al (2016) A trans-synaptic nanocolumn aligns neurotransmitter release to receptors. Nature 536:210–214
Tatavarty V, Kim E-J, Rodionov V, Yu J (2009) Investigating sub-spine actin dynamics in rat hippocampal neurons with super-resolution optical imaging. PLoS ONE 4:e7724
Testa I, Urban NT, Jakobs S et al (2012) Nanoscopy of living brain slices with low light levels. Neuron 75:992–1000
Tillberg PW, Chen F, Piatkevich KD et al (2016) Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteinsand antibodies. Nat Biotechnol 34:987–992
Tønnesen J, Nägerl UV (2016) Dendritic spines as tunable regulators of synaptic signals. Front Psychiatry 7:101
Tønnesen J, Katona G, Rózsa B, Nägerl UV (2014) Spine neck plasticity regulates compartmentalization of synapses. Nat Neurosci 17:678–685
Tønnesen J, Inavalli VVGK, Nägerl UV (2018) Super-resolution imaging of the extracellular space in living brain tissue. Cell 172:1108-1121.e15
Trotter JH, Hao J, Maxeiner S et al (2019) Synaptic neurexin-1 assembles into dynamically regulated active zone nano-clusters. J Cell Biol 218:2677–2698
Truckenbrodt S, Maidorn M, Crzan D et al (2018) X10 expansion microscopy enables 25-nm resolution on conventional microscopes. EMBO Rep. https://doi.org/10.15252/embr.201845836
Turcotte R, Liang Y, Tanimoto M et al (2019) Dynamic super-resolution structured illumination imaging in the living brain. Proc Natl Acad Sci USA 116:9586–9591
Urban NT, Willig KI, Hell SW, Nägerl UV (2011) STED nanoscopy of actin dynamics in synapses deep inside living brain slices. Biophys J 101:1277–1284
Vicidomini G, Bianchini P, Diaspro A (2018) STED super-resolved microscopy. Nat Methods 15:173–182
Weeks ACW, Ivanco TL, Leboutillier JC et al (1999) Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus: I. Intermed Maint Phase Synap 31:97–107
Wegner W, Ilgen P, Gregor C et al (2017) In vivo mouse and live cell STED microscopy of neuronal actin plasticity using far-red emitting fluorescent proteins. Sci Rep 7:1–10
Wegner W, Mott AC, Grant SGN et al (2018) In vivo STED microscopy visualizes PSD95 sub-structures and morphological changes over several hours in the mouse visual cortex. Sci Rep 8:219
Wilhelm BG, Mandad S, Truckenbrodt S et al (2014) Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins. Science 344:1023–1028
Willig KI, Steffens H, Gregor C et al (2014) Nanoscopy of filamentous actin in cortical dendrites of a living mouse. Biophys J 106:L01–L03
Wu Y, Shroff H (2018) Faster, sharper, and deeper: structured illumination microscopy for biological imaging. Nat Methods 15:1011–1019
Xu K, Zhong G, Zhuang X (2013) Actin, spectrin, and associated proteins form a periodic cytoskeletal structure in axons. Science 339:452–456
York AG, Parekh SH, Dalle Nogare D et al (2012) Resolution doubling in live, multicellular organisms via multifocal structured illumination microscopy. Nat Methods 9:749–754
York AG, Chandris P, Nogare DD et al (2013) Instant super-resolution imaging in live cells and embryos via analog image processing. Nat Methods 10:1122–1130
Yuste R (2013) Electrical compartmentalization in dendritic spines. Annu Rev Neurosci 36:429–449
Zaccard CR, Shapiro L, Martin-de-Saavedra MD et al (2020) Rapid 3D enhanced resolution microscopy reveals diversity in dendritic spinule dynamics, regulation, and function. Neuron 107:522–537
Acknowledgements
We thank Dr. Kazuki Obashi (National Institutes of Health, Bethesda, USA) for his valuable comments. This work was supported by JSPS KAKENHI (20K15892 to Y. K.).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests exist.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Kashiwagi, Y., Okabe, S. Imaging of spine synapses using super-resolution microscopy. Anat Sci Int 96, 343–358 (2021). https://doi.org/10.1007/s12565-021-00603-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12565-021-00603-0