Abstract
Neurons transform the information arising from up to thousands of synaptic inputs into specific spiking patterns. Nonlinear dendritic integration is a crucial step in this process and is thought to increase the computational ability of neurons. However, studying how complex spatiotemporal patterns of synaptic inputs drive neuronal spiking is technically challenging. Two-photon neurotransmitter uncaging allows researchers to activate sequences of single synapses with high spatiotemporal precision and thus systematically examine how single and multiple synaptic activation patterns may recruit dendritic nonlinearities. Here, we describe the theoretical and practical considerations of using two-photon uncaging to mimic synaptic activation and monitor the electrical and biochemical signaling in dendrites when evoked by various synaptic patterns.
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Abbreviations
- [Glut]:
-
Glutamate concentration
- 2PLSM:
-
Two-photon laser scanning microscopy
- ACSF:
-
Artificial cerebrospinal fluid
- AMPAR:
-
α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor
- AOD:
-
Acousto-optic deflector
- AOM:
-
Acousto-optic modulator
- CDNI-glutamate:
-
4-Carboxymethoxy-5,7-Dinitroindolinyl-Glutamate
- dI/O:
-
Dendritic input/output relationship
- DiO/DPA:
-
Two-component optical voltage sensor made of the neuronal tracer dye DiOC16(3) and dipicrylamine
- DNI-glutamate TFA:
-
4-Methoxy-5,7-dinitroindolinyl-L-glutamate trifluoroacetate
- GM:
-
The molecular two-photon cross-section is usually quoted in the units of Göppert-Mayer (GM), where 1 GM is 10−50 cm4 s photon−1
- I/O:
-
Input/output relationship
- MNI-Glutamate:
-
4-Methoxy-7-nitroindolinyl-caged-L-glutamate
- NMDAR:
-
N-Methyl-D-aspartate receptor
- PSF:
-
Point spread function
- PV:
-
Parvalbumin
- SLM:
-
Spatial light modulator
- u[Glut]:
-
Uncaging-evoked glutamate transient
- uEPSC:
-
Uncaging-evoked EPSC
- uEPSP:
-
Uncaging-evoked EPSP
- VGCC:
-
Voltage-gated calcium channel
- θ B :
-
Bragg angle, the angle between the incident laser beam into an AOM and the diffracted beam
References
Rall W (1967) Distinguishing theoretical synaptic potentials computed for different soma-dendritic distributions of synaptic input. J Neurophysiol 30:1138–1168
Magee JC, Johnston D (1995) Characterization of single voltage-gated Na+ and Ca2+ channels in apical dendrites of rat CA1 pyramidal neurons. J Physiol 487:67–90. https://doi.org/10.1113/jphysiol.1995.sp020862
Stuart GJ, Sakmann B (1994) Active propagation of somatic action potentials into neocortical pyramidal cell dendrites. Nature 367:69–72. https://doi.org/10.1038/367069a0
Williams SR, Mitchell SJ (2008) Direct measurement of somatic voltage clamp errors in central neurons. Nat Neurosci 11:790–798. https://doi.org/10.1038/nn.2137
Corrie JET (2005) Photoremovable protecting groups used for the caging of biomolecules. Dyn Stud Biol 1999:1–94
Lester HA, Nerbonne JM (1982) Physiological and pharmacological manipulations with light flashes. Annu Rev Biophys Bioeng 11:151–175. https://doi.org/10.1146/annurev.bb.11.060182.001055
Sarkisov DV, Wang SS-H (2007) Combining uncaging techniques with patch-clamp recording and optical physiology. In: Walz W (ed) Neuromethods, Patch-Clamp analysis, vol 38, 2nd edn. Humana Press, Totowa, NJ, pp 149–168. https://doi.org/10.1007/978-1-59745-492-6_5
Palma-Cerda F, Auger C, Crawford DJ et al (2012) New caged neurotransmitter analogs selective for glutamate receptor sub-types based on methoxynitroindoline and nitrophenylethoxycarbonyl caging groups. Neuropharmacology 63:624–634. https://doi.org/10.1016/j.neuropharm.2012.05.010
Fino E, Araya R, Peterka DS et al (2009) RuBi-Glutamate: two-photon and visible-light photoactivation of neurons and dendritic spines. Front Neural Circuits 3:1–9. https://doi.org/10.3389/neuro.04.002.2009
Zipfel WR, Williams RM, Webb WW (2003) Nonlinear magic: multiphoton microscopy in the biosciences. Nat Biotechnol 21:1369–1377. https://doi.org/10.1038/nbt899
Trigo FF, Corrie JET, Ogden D (2009) Laser photolysis of caged compounds at 405 nm: photochemical advantages, localisation, phototoxicity and methods for calibration. J Neurosci Meth 180:9–21. https://doi.org/10.1016/j.jneumeth.2009.01.032
Carter AG, Sabatini BL (2004) State-dependent calcium signaling in dendritic spines of striatal medium spiny neurons. Neuron 44:483–493. https://doi.org/10.1016/j.neuron.2004.10.013
Losonczy A, Magee JC (2006) Integrative properties of radial oblique dendrites in hippocampal CA1 pyramidal neurons. Neuron 50:291–307. https://doi.org/10.1016/j.neuron.2006.03.016
Branco T, Häusser M (2011) Synaptic integration gradients in single cortical pyramidal cell dendrites. Neuron 69:885–892. https://doi.org/10.1016/j.neuron.2011.02.006
Tran-Van-Minh A, Abrahamsson T, Cathala L, DiGregorio DA (2016) Differential dendritic integration of synaptic potentials and calcium in cerebellar interneurons. Neuron 91:837–850. https://doi.org/10.1016/j.neuron.2016.07.029
Salierno M, Marceca E, Peterka DS et al (2010) A fast ruthenium polypyridine cage complex photoreleases glutamate with visible or IR light in one and two photon regimes. J Inorg Biochem 104:418–422. https://doi.org/10.1016/j.jinorgbio.2009.12.004
Ellis-Davies GCR, Matsuzaki M, Paukert M et al (2007) 4-Carboxymethoxy-5,7-Dinitroindolinyl-Glu: an improved caged glutamate for expeditious ultraviolet and two-photon photolysis in brain slices. J Neurosci 27:6601–6604. https://doi.org/10.1523/JNEUROSCI.1519-07.2007
Chiovini B, Turi GF, Katona G et al (2014) Dendritic spikes induce ripples in parvalbumin interneurons during hippocampal sharp waves. Neuron 82:908–924. https://doi.org/10.1016/j.neuron.2014.04.004
Gasparini S, Magee JC (2006) State-dependent dendritic computation in hippocampal CA1 pyramidal neurons. J Neurosci 26:2088–2100. https://doi.org/10.1523/JNEUROSCI.4428-05.2006
Fernandez-Alfonso T, Nadella KMNS, Iacaruso MF et al (2014) Monitoring synaptic and neuronal activity in 3D with synthetic and genetic indicators using a compact acousto-optic lens two-photon microscope. J Neurosci Meth 222:69–81. https://doi.org/10.1016/j.jneumeth.2013.10.021
Katona G, Szalay G, Maák P et al (2012) Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes. Nat Methods 9:201–208. https://doi.org/10.1038/nmeth.1851
Lutz C, Otis TS, DeSars V et al (2008) Holographic photolysis of caged neurotransmitters. Nat Methods 5:821–827. https://doi.org/10.1038/nmeth.1241
Hernandez O, Papagiakoumou E, Tanese D et al (2016) Three-dimensional spatiotemporal focusing of holographic patterns. Nat Commun 7:12716. https://doi.org/10.1038/ncomms12716
Anselmi F, Ventalon C, Begue A et al (2011) Three-dimensional imaging and photostimulation by remote-focusing and holographic light patterning. Proc Natl Acad Sci USA 108:19504–19509. https://doi.org/10.1073/pnas.1109111108
Bovetti S, Fellin T (2015) Optical dissection of brain circuits with patterned illumination through the phase modulation of light. J Neurosci Meth 241:66–77. https://doi.org/10.1016/j.jneumeth.2014.12.002
Smirnov MS, Evans PR, Garrett TR et al (2017) Automated remote focusing, drift correction, and photostimulation to evaluate structural plasticity in dendritic spines. PLoS One 12:1–14. https://doi.org/10.1371/journal.pone.0170586
Matsuzaki M, Ellis-Davies GC, Nemoto T et al (2001) Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons. Nat Neurosci 4:1086–1092. https://doi.org/10.1038/nn736
Jonas P (2000) The time course of signaling at central glutamatergic synapses. News Physiol Sci 15:83–89. https://doi.org/10.1113/jphysiol.1992.sp019417
Crank J (1975) The mathematics of diffusion, 2nd edn. Oxford University Press, Oxford
Nielsen TA, DiGregorio DA, Silver RA (2004) Modulation of glutamate mobility reveals the mechanism underlying slow-rising AMPAR EPSCs and the diffusion coefficient in the synaptic cleft. Neuron 42:757–771. https://doi.org/10.1016/j.neuron.2004.04.003
DiGregorio DA, Rothman JS, Nielsen TA, Silver RA (2007) Desensitization properties of AMPA receptors at the cerebellar mossy fiber granule cell synapse. J Neurosci 27:8344–8357. https://doi.org/10.1523/JNEUROSCI.2399-07.2007
Kiskin NI, Chillingworth R, McCray JA et al (2002) The efficiency of two-photon photolysis of a “caged” fluorophore, o-1-(2-nitrophenyl)ethylpyranine, in relation to photodamage of synaptic terminals. Eur Biophys J 30:588–604. https://doi.org/10.1007/s00249-001-0187-x
Koester HJ, Baur D, Uhl R, Hell SW (1999) Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage. Biophys J 77:2226–2236. https://doi.org/10.1016/S0006-3495(99)77063-3
Axelrod D, Koppel DE, Schlessinger J et al (1976) Mobility measurement by analysis of fluorescence recovery kinetics. Biophys J 16:1055–1069
Kiskin NI, Ogden D (2002) Two-photon excitation and photolysis by pulsed laser illumination modelled by spatially non-uniform reactions with simultaneous diffusion. Eur Biophys J 30:571–587. https://doi.org/10.1007/s00249-001-0186-y
McBain CJ, Fisahn A (2001) Interneurons unbound. Nat Rev Neurosci 2:11–23. https://doi.org/10.1038/35049047
Nevian T, Larkum ME, Polsky A, Schiller J (2007) Properties of basal dendrites of layer 5 pyramidal neurons: a direct patch-clamp recording study. Nat Neurosci 10:206–214. https://doi.org/10.1038/nn1826
Magee JC, Cook EP (2000) Somatic EPSP amplitude is independent of synapse location in hippocampal pyramidal neurons. Nat Neurosci 3:895–903
Smith MA, Ellis-Davies GCR, Magee JC (2003) Mechanism of the distance-dependent scaling of Schaffer collateral synapses in rat CA1 pyramidal neurons. J Physiol 548:245–258. https://doi.org/10.1113/jphysiol.2002.036376
Geiger JRP, Lübke J, Roth A et al (1997) Submillisecond AMPA receptor-mediated signalling at a principal neuron-interneuron synapse. Neuron 18:1009–1023
Cane M, Maco B, Knott G, Holtmaat A (2014) The relationship between PSD-95 clustering and spine stability in vivo. J Neurosci 34:2075–2086. https://doi.org/10.1523/JNEUROSCI.3353-13.2014
Abrahamsson T, Cathala L, Matsui K et al (2012) Thin dendrites of cerebellar interneurons confer sublinear synaptic integration and a gradient of short-term plasticity. Neuron 73:1159–1172. https://doi.org/10.1016/j.neuron.2012.01.027
Fortin DA, Tillo SE, Yang G et al (2014) Live imaging of endogenous PSD-95 using ENABLED: a conditional strategy to fluorescently label endogenous proteins. J Neurosci 34:16698–16712. https://doi.org/10.1523/JNEUROSCI.3888-14.2014
Tønnesen J, Katona G, Rózsa B, Nägerl UV (2014) Spine neck plasticity regulates compartmentalization of synapses. Nat Neurosci 17:678–685. https://doi.org/10.1038/nn.3682
Popovic MA, Carnevale N, Rozsa B, Zecevic D (2015) Electrical behaviour of dendritic spines as revealed by voltage imaging. Nat Commun 6:8436. https://doi.org/10.1038/ncomms9436
Acker CD, Hoyos E, Loew LM (2016) EPSPs measured in proximal dendritic spines of cortical pyramidal neurons. eNeuro 3:1–13. https://doi.org/10.1523/ENEURO.0050-15.2016
Bradley J, Luo R, Otis TS, DiGregorio DA (2009) Submillisecond optical reporting of membrane potential in situ using a neuronal tracer dye. J Neurosci 29:9197–9209. https://doi.org/10.1523/JNEUROSCI.1240-09.2009
Fink AE, Bender KJ, Trussell LO et al (2012) Two-photon compatibility and single-voxel, single-trial detection of subthreshold neuronal activity by a two-component optical voltage sensor. PLoS One 7. https://doi.org/10.1371/journal.pone.0041434
Palmer LM, Stuart GJ (2009) Membrane potential changes in dendritic spines during action potentials and synaptic input. J Neurosci 29:6897–6903. https://doi.org/10.1523/JNEUROSCI.5847-08.2009
Krueppel R, Remy S, Beck H (2011) Dendritic integration in hippocampal dentate granule cells. Neuron 71:512–528. https://doi.org/10.1016/j.neuron.2011.05.043
Poirazi P, Brannon T, Mel BW (2003) Pyramidal neuron as two-layer neural network. Neuron 37:989–999. https://doi.org/10.1016/S0896-6273(03)00149-1
Hu H, Martina M, Jonas P (2010) Dendritic mechanisms underlying rapid synaptic activation of fast-spiking hippocampal interneurons. Science 327:52–58. https://doi.org/10.1126/science.1177876
Branco T, Clark BA, Häusser M (2010) Dendritic discrimination of temporal input sequences in cortical neurons. Science 329:1671–1675. https://doi.org/10.1126/science.1189664
Agmon-Snir H, Carr CE, Rinzel J (1998) The role of dendrites in auditory coincidence detection. Nature 393:268–272. https://doi.org/10.1038/30505
Mel BW, Ruderman DL, Archie KA (1998) Translation-invariant orientation tuning in visual “complex” cells could derive from intradendritic computations. J Neurosci 18:4325–4334. https://doi.org/10.1098/rspb.1986.0060
Losonczy A, Makara JK, Magee JC (2008) Compartmentalized dendritic plasticity and input feature storage in neurons. Nature 452:436–441. https://doi.org/10.1038/nature06725
Xu N, Harnett MT, Williams SR et al (2012) Nonlinear dendritic integration of sensory and motor input during an active sensing task. Nature 492:247–251. https://doi.org/10.1038/nature11601
Schmidt-Hieber C, Toleikyte G, Aitchison L et al (2017) Active dendritic integration as a mechanism for robust and precise grid cell firing. Nat Neurosci 20:1114–1121. https://doi.org/10.1038/nn.4582
Wilson DE, Whitney DE, Scholl B, Fitzpatrick D (2016) Orientation selectivity and the functional clustering of synaptic inputs in primary visual cortex. Nat Neurosci 19:1003–1009. https://doi.org/10.1038/nn.4323
Scholl B, Wilson DE, Fitzpatrick D (2017) Local order within global disorder: synaptic architecture of visual space. Neuron 96:1127–1138. https://doi.org/10.1016/j.neuron.2017.10.017
Soler-Llavina GJ, Sabatini BL (2006) Synapse-specific plasticity and compartmentalized signaling in cerebellar stellate cells. Nat Neurosci 9:798–806. https://doi.org/10.1038/nn1698
Weber JP, Andrásfalvy BK, Polito M et al (2016) Location-dependent synaptic plasticity rules by dendritic spine cooperativity. Nat Commun 7:11380. https://doi.org/10.1038/ncomms11380
Silver RA (2010) Neuronal arithmetic. Nat Rev Neurosci 11:474–489. https://doi.org/10.1038/nrn2864
Tran-Van-Minh A, Cazé RD, Abrahamsson T et al (2015) Contribution of sublinear and supralinear dendritic integration to neuronal computations. Front Cell Neurosci 9:1–15. https://doi.org/10.3389/fncel.2015.00067
Higley MJ, Soler-Llavina GJ, Sabatini BL (2009) Cholinergic modulation of multivesicular release regulates striatal synaptic potency and integration. Nat Neurosci 12:1121–1128. https://doi.org/10.1038/nn.2368
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Tran-Van-Minh, A., Rebola, N., Hoehne, A., DiGregorio, D.A. (2019). Two-Photon Neurotransmitter Uncaging for the Study of Dendritic Integration. In: Hartveit, E. (eds) Multiphoton Microscopy. Neuromethods, vol 148. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9702-2_3
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DOI: https://doi.org/10.1007/978-1-4939-9702-2_3
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