Abstract
Information is delivered to neurons through thousands of synapses, most of them located on the branches of treelike structures we call dendrites. These dendrites are quite remarkable since on them lies the responsibility of collecting most of the inputs to a neuron and channeling this information to the soma and axon hillock where a decision is made to fire or not an action potential, to participate or not in the active circuit.
Dendrites are therefore the place where structure interacts with information in a neuron. Since dendrites are not sphere-like structures, which would render the neurons iso-potential units, but look like a tree, their morphology and the distribution of synapses on their branches reflects the nature of this interaction and therefore are some of the most crucial components defining the computations performed within each neuron through development to adulthood. This makes dendritic structure and connectivity two of the most important defining features of cell types.
Electron microscopy (EM) is perfectly poised to investigate connectivity as well as the fine morphology of the dendrites since it is widely recognized as the gold standard to identify synapses and can image at the resolution to precisely quantify their morphology and if required their intracellular organelles and components.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ahmed B, Anderson J, Douglas R et al (1994) Polyneuronal innervation of spiny stellate neurons in cat visual cortex. J Comp Neurol 341:39–49
Ahmed B, Anderson J, Martin K, Nelson J (1997) Map of the synapses onto layer 4 basket cells of the primary visual cortex of the cat. J Comp Neurol 380:230–242
Albertson DG, Thomson JN (1976) The pharynx of Caenorhabditis elegans. Philos Trans R Soc Lond B Biol Sci 275:299–325
Anderson J, Douglas R, Martin K, Nelson J (1994) Map of the synapses formed with the dendrites of spiny stellate neurons of cat visual cortex. J Comp Neurol 341:25–38
Atasoy D, Betley JN, Li W-P et al (2014) A genetically specified connectomics approach applied to long-range feeding regulatory circuits. Nat Neurosci 17:1830–1839. doi:10.1038/nn.3854
Banitt Y, Martin K, Segev I (2007) A biologically realistic model of contrast invariant orientation tuning by thalamocortical synaptic depression. J Neurosci 27:10230–10239. doi:10.1523/JNEUROSCI.1640-07.2007
Beaulieu C (1993) Numerical data on neocortical neurons in adult rat, with special reference to the GABA population. Brain Res 609:284–292
Beaulieu C, Kisvarday Z, Somogyi P et al (1992) Quantitative distribution of GABA-immunopositive and-immunonegative neurons and synapses in the monkey striate cortex (area 17). Cereb Cortex 2:295–309
Binzegger T, Douglas R, Martin K (2004) A quantitative map of the circuit of cat primary visual cortex. J Neurosci 24:8441–8453
Blackstad TW (1965) Mapping of experimental axon degeneration by electron microscopy of Golgi preparations. Z Zellforsch Mikrosk Anat 67:819–834
Blackstad TW (1975) Electron microscopy of experimental axonal degeneration in photochemically modified Golgi preparations: a procedure for precise mapping of nervous connections. Brain Res 95:191–210
Bleckert A, Parker ED, Kang Y et al (2013) Spatial relationships between GABAergic and glutamatergic synapses on the dendrites of distinct types of mouse retinal ganglion cells across development. PLoS One 8:e69612. doi:10.1371/journal.pone.0069612
Bock DD, Lee W-CA, Kerlin AM et al (2011) Network anatomy and in vivo physiology of visual cortical neurons. Nature 471:177–182. doi:10.1038/nature09802
Braitenberg V, Schüz A (1991) Anatomy of the cortex, 2nd edn. Springer, Berlin
Briggman KL, Bock DD (2012) Volume electron microscopy for neuronal circuit reconstruction. Curr Opin Neurobiol 22:154–161. doi:10.1016/j.conb.2011.10.022
Briggman KL, Helmstaedter M, Denk W (2011) Wiring specificity in the direction-selectivity circuit of the retina. Nature 471:183–188. doi:10.1038/nature09818
Bumbarger DJ, Riebesell M, Rödelsperger C, Sommer RJ (2013) System-wide rewiring underlies behavioral differences in predatory and bacterial-feeding nematodes. Cell 152:109–119. doi:10.1016/j.cell.2012.12.013
Chen Y et al (2012) Signaling in dendritic spines and spine microdomains. Curr Opin Neurobiol 22:389
Colonnier M (1968) Synaptic patterns on different cell types in the different laminae of the cat visual cortex. An electron microscope study. Brain Res 9:268–287
Cragg B (1967) The density of synapses and neurones in the motor and visual areas of the cerebral cortex. J Anat 101:639–654
Cuntz H, Borst A, Segev I (2007) Optimization principles of dendritic structure. Theor Biol Med Model 4:21. doi:10.1186/1742-4682-4-21
Da Costa NM (2013) Diversity of thalamorecipient spine morphology in cat visual cortex and its implication for synaptic plasticity. J Comp Neurol 521:2058–2066. doi:10.1002/cne.23272
Da Costa NM, Martin KAC (2011) How thalamus connects to spiny stellate cells in the cat’s visual cortex. J Neurosci 31:2925–2937. doi:10.1523/JNEUROSCI.5961-10.2011
Da Costa NM, Martin KAC (2013) Sparse reconstruction of brain circuits: or, how to survive without a microscopic connectome. NeuroImage:1–10. doi:10.1016/j.neuroimage.2013.04.054
Davis T, Sterling P (1979) Microcircuitry of cat visual cortex: classification of neurons in layer IV of area 17, and identification of the patterns of lateral geniculate input. J Comp Neurol 188:599–627
Denk W, Horstmann H (2004) Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure. PLoS Biol 2:e329
Fiala JC, Harris KM (1999) Dendrite structure. In: Stuart G, Spruston N, Häusser M (eds) Dendrites. Oxford University Press, Oxford
Gong S, Doughty M, Harbaugh CR et al (2007) Targeting Cre recombinase to specific neuron populations with bacterial artificial chromosome constructs. J Neurosci 27:9817–9823. doi:10.1523/JNEUROSCI.2707-07.2007
Gray EG (1959a) Axo-somatic and axo-dendritic synapses of the cerebral cortex: an electron microscope study. J Anat 93:420–433
Gray EG (1959b) Electron microscopy of synaptic contacts on dendrite spines of the cerebral cortex. Nature 183:1592–1593
Harris KM, Stevens JK (1988) Dendritic spines of rat cerebellar Purkinje cells: serial electron microscopy with reference to their biophysical characteristics. J Neurosci 8:4455–4469
Harris KM, Stevens JK (1989) Dendritic spines of CA 1 pyramidal cells in the rat hippocampus: serial electron microscopy with reference to their biophysical characteristics. J Neurosci 9:2982–2997
Harris JA, Hirokawa KE, Sorensen SA et al (2014) Anatomical characterization of Cre driver mice for neural circuit mapping and manipulation. Front Neural Circ 8:76. doi:10.3389/fncir.2014.00076
Hayworth KJ, Morgan JL, Schalek R et al (2014) Imaging ATUM ultrathin section libraries with WaferMapper: a multi-scale approach to EM reconstruction of neural circuits. Front Neural Circ 8:68. doi:10.3389/fncir.2014.00068
Helmstaedter M, Briggman KL, Turaga SC et al (2013) Connectomic reconstruction of the inner plexiform layer in the mouse retina. Nature 500:168–174. doi:10.1038/nature12346
Jaffe DB, Carnevale NT (1999) Passive normalization of synaptic integration influenced by dendritic architecture. J Neurophysiol 82:3268–3285
Jarrell TA, Wang Y, Bloniarz AE et al (2012) The connectome of a decision-making neural network. Science 337:437–444. doi:10.1126/science.1221762
Jones EG (2007) Thalamic neurons, synaptic organization, and functional properties. In: The Thalamus, 2nd edn. Cambridge University Press, Cambridge, pp 192–204
Kisvárday ZF, Martin KA, Freund TF et al (1986) Synaptic targets of HRP-filled layer III pyramidal cells in the cat striate cortex. Exp Brain Res 64:541–552
Knott GW, Holtmaat A, Trachtenberg JT et al (2009) A protocol for preparing GFP-labeled neurons previously imaged in vivo and in slice preparations for light and electron microscopic analysis. Nat Protoc 4:1145–1156. doi:10.1038/nprot.2009.114
Koch C, Zador A (1993) The function of dendritic spines: devices subserving biochemical rather than electrical compartmentalization. J Neurosci 13:413–422
Kubota Y, Karube F, Nomura M et al (2011) Conserved properties of dendritic trees in four cortical interneuron subtypes. Sci Rep 1:89. doi:10.1038/srep00089
Lam SS, Martell JD, Kamer KJ et al (2015) Directed evolution of APEX2 for electron microscopy and proximity labeling. Nat Methods 12:51–54. doi:10.1038/nmeth.3179
Lefort S, Tomm C, Sarria JF, Petersen C (2009) The excitatory neuronal network of the C2 barrel column in mouse primary somatosensory cortex. Neuron 61:301
Lichtman JW, Pfister H, Shavit N (2014) The big data challenges of connectomics. Nat Neurosci 17:1448–1454. doi:10.1038/nn.3837
London M, Häusser M (2005) Dendritic computation. Annu Rev Neurosci 28:503–532. doi:10.1146/annurev.neuro.28.061604.135703
Maco B, Holtmaat A, Cantoni M et al (2013) Correlative in vivo 2 photon and focused ion beam scanning electron microscopy of cortical neurons. PLoS One 8:e57405. doi:10.1371/journal.pone.0057405
Mainen ZF, Sejnowski TJ (1996) Influence of dendritic structure on firing pattern in model neocortical neurons. Nature 382:363–366. doi:10.1038/382363a0
Martell JD, Deerinck TJ, Sancak Y et al (2012) Engineered ascorbate peroxidase as a genetically encoded reporter for electron microscopy. Nat Biotechnol 30:1143–1148. doi:10.1038/nbt.2375
Micheva KD, Smith SJ (2007) Array tomography: a new tool for imaging the molecular architecture and ultrastructure of neural circuits. Neuron 55:25–36. doi:10.1016/j.neuron.2007.06.014
Micheva KD, Busse B, Weiler NC et al (2010) Single-synapse analysis of a diverse synapse population: proteomic imaging methods and markers. Neuron 68:639–653. doi:10.1016/j.neuron.2010.09.024
Miki T, Fukui Y, Itoh M et al (1997) Estimation of the numerical densities of neurons and synapses in cerebral cortex. Brain Res Brain Res Protoc 2:9–16
O’Kusky J, Colonnier M (1982) A laminar analysis of the number of neurons, glia, and synapses in the adult cortex (area 17) of adult macaque monkeys. J Comp Neurol 210:278–290. doi:10.1002/cne.902100307
Paez-Segala MG, Sun MG, Shtengel G et al (2015) Fixation-resistant photoactivatable fluorescent proteins for CLEM. Nat Methods. doi:10.1038/nmeth.3225
Peters A, Feldman M (1977) The projection of the lateral geniculate nucleus to area 17 of the rat cerebral cortex. IV. Terminations upon spiny dendrites. J Neurocytol 6:669–689
Peters A, Harriman KM (1990) Different kinds of axon terminals forming symmetric synapses with the cell bodies and initial axon segments of layer II/III pyramidal cells. I. Morphometric analysis. J Neurocytol 19:154–174
Peters A, Harriman KM (1992) Different kinds of axon terminals forming symmetric synapses with the cell bodies and initial axon segments of layer II/III pyramidal cells. III. Origins and frequency of occurrence of the terminals. J Neurocytol 21:679–692
Peters A, Kaiserman-Abramof I (1970) The small pyramidal neuron of the rat cerebral cortex. The perikaryon, dendrites and spines. Am J Anat 127:321–355
Peters A, Saint Marie RL (1984) Smooth and sparsely spinous nonpyramidal cells forming local axonal plexuses. In: Peters A, Jones EG (eds) Cerebral cortex, vol 1. Cellular components of the cerebral cortex. Plenum, New York, pp 419
Peters A, Feldman M, Saldanha J (1976) The projection of the lateral geniculate nucleus to area 17 of the rat cerebral cortex. II. Terminations upon neuronal perikarya and dendritic shafts. J Neurocytol 5:85–107
Peters A, Sethares C, Harriman KM (1990) Different kinds of axon terminals forming symmetric synapses with the cell bodies and initial axon segments of layer II/III pyramidal cells. II. Synaptic junctions. J Neurocytol 19:584–600
Peters A, Palay S, Webster F (1991) The fine structure of the nervous system: neurons and their supporting cells, 3rd edn. Oxford University Press, New York
Plaza SM, Scheffer LK, Chklovskii DB (2014) Toward large-scale connectome reconstructions. Curr Opin Neurobiol 25:201–210. doi:10.1016/j.conb.2014.01.019
Rall W (1959) Branching dendritic trees and motoneuron membrane resistivity. Exp Neurol 1:491–527. doi:10.1016/0014-4886(59)90046-9
Rall W (1964) Theoretical significance of dendritic trees for neuronal input-output relations. In Reiss RF (ed) Neural theory and modeling. Stanford University Press, Stanford, pp 73–97
Saint Marie R, Peters A (1985) The morphology and synaptic connections of spiny stellate neurons in monkey visual cortex (area 17): a Golgi-electron microscopic study. J Comp Neurol 233:213–235
Schoonover CE, Tapia J-C, Schilling VC et al (2014) Comparative strength and dendritic organization of thalamocortical and corticocortical synapses onto excitatory layer 4 neurons. J Neurosci 34:6746–6758. doi:10.1523/JNEUROSCI.0305-14.2014
Schüz A, Palm G (1989) Density of neurons and synapses in the cerebral cortex of the mouse. J Comp Neurol 286:442–455. doi:10.1002/cne.902860404
Segev I, London M (2000) Untangling dendrites with quantitative models. Science 290:744–750
Segev I, Friedman A, White E, Gutnick M (1995a) Electrical consequences of spine dimensions in a model of a cortical spiny stellate cell completely reconstructed from serial thin sections. J Comput Neurosci 2:117–130
Segev I, Shepherd G, Rinzel J (1995b) The theoretical foundation of dendritic functions. Bradford Book, The MIT Press, Cambridge, Mass
Shu X, Lev-Ram V, Deerinck TJ et al (2011) A genetically encoded tag for correlated light and electron microscopy of intact cells, tissues, and organisms. PLoS Biol 9:e1001041. doi:10.1371/journal.pbio.1001041.g005
Somogyi P, Kisvárday ZF, Martin KA, Whitteridge D (1983) Synaptic connections of morphologically identified and physiologically characterized large basket cells in the striate cortex of cat. Neuroscience 10:261–294
Stell WK (1965) Correlation of retinal cytoarchitecture and ultrastructure in Golgi preparations. Anat Rec 153:389–397
Takemura S-Y, Bharioke A, Lu Z et al (2013) A visual motion detection circuit suggested by Drosophila connectomics. Nature 500:175–181. doi:10.1038/nature12450
Taniguchi H, He M, Wu P et al (2011) A resource of Cre driver lines for genetic targeting of GABAergic neurons in cerebral cortex. Neuron 71:995–1013. doi:10.1016/j.neuron.2011.07.026
Turner AM, Greenough WT (1985) Differential rearing effects on rat visual cortex synapses. I. Synaptic and neuronal density and synapses per neuron. Brain Res 329:195–203
Vaughan DW, Peters A (1973) A three dimensional study of layer I of the rat parietal cortex. J Comp Neurol 149:355–370. doi:10.1002/cne.901490305
Wang H-P, Spencer D, Fellous J-M, Sejnowski TJ (2010) Synchrony of thalamocortical inputs maximizes cortical reliability. Science 328:106–109. doi:10.1126/science.1183108
Warren MA, Bedi KS (1982) Synapse-to-neuron ratios in the visual cortex of adult rats undernourished from about birth until 100 days of age. J Comp Neurol 210:59–64. doi:10.1002/cne.902100107
White E, Rock M (1979) Distribution of thalamic input to different dendrites of a spiny stellate cell in mouse sensorimotor cortex. Neurosci Lett 15:115–119
White E, Rock M (1980) Three-dimensional aspects and synaptic relationships of a Golgi-impregnated spiny stellate cell reconstructed from serial thin sections. J Neurocytol 9:615–636
White JG, Southgate E, Thomson JN, Brenner S (1976) The structure of the ventral nerve cord of Caenorhabditis elegans. Philos Trans R Soc Lond B Biol Sci 275:327–348
White J, Southgate E, Thomson J, Brenner S (1986) The structure of the nervous system of the nematode Caenorhabditis elegans. Philos Trans R Soc Lond B Biol Sci 314:1–340
Wikipedia Morphology (linguistics). In: httpen.wikipedia.orgwikiMorphologylinguistics. Accessed 20 Jan 2015
Yuste R (2011) Dendritic spines and distributed circuits. Neuron 71:772–781. doi:10.1016/j.neuron.2011.07.024
Yuste R, Denk W (1995) Dendritic spines as basic functional units of neuronal integration. Nature 375:682–684. doi:10.1038/375682a0
Acknowledgments
I would like to thank Agnes Bodor, JoAnn Buchanan, and Marc Takeno for their help preparing the figures and to Agnes Bodor, Hannah Krakauer, Jill Scott, and Elizabeth Whitney for their comments on the manuscript. I also would like to thank Ali Cetin and Tanya Daigle-Ting for the generation and optimization of Apex expressing recombinant adeno-associated viral vectors for in vivo applications. I am grateful to the Allen Institute founders, P.G. Allen and J. Allen, for their vision, encouragement, and support.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Japan
About this chapter
Cite this chapter
da Costa, N.M. (2016). Ultrastructure Methods. In: Emoto, K., Wong, R., Huang, E., Hoogenraad, C. (eds) Dendrites. Springer, Tokyo. https://doi.org/10.1007/978-4-431-56050-0_6
Download citation
DOI: https://doi.org/10.1007/978-4-431-56050-0_6
Published:
Publisher Name: Springer, Tokyo
Print ISBN: 978-4-431-56048-7
Online ISBN: 978-4-431-56050-0
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)