Abstract
Understanding cortical circuits will require mapping the connections between specific populations of neurons1, as well as determining the dendritic locations where the synapses occur2. The dendrites of individual cortical neurons overlap with numerous types of local and long-range excitatory axons, but axodendritic overlap is not always a good predictor of actual connection strength3,4,5. Here we developed an efficient channelrhodopsin-2 (ChR2)-assisted method6,7,8 to map the spatial distribution of synaptic inputs, defined by presynaptic ChR2 expression, within the dendritic arborizations of recorded neurons. We expressed ChR2 in two thalamic nuclei, the whisker motor cortex and local excitatory neurons and mapped their synapses with pyramidal neurons in layers 3, 5A and 5B (L3, L5A and L5B) in the mouse barrel cortex. Within the dendritic arborizations of L3 cells, individual inputs impinged onto distinct single domains. These domains were arrayed in an orderly, monotonic pattern along the apical axis: axons from more central origins targeted progressively higher regions of the apical dendrites. In L5 arborizations, different inputs targeted separate basal and apical domains. Input to L3 and L5 dendrites in L1 was related to whisker movement and position, suggesting that these signals have a role in controlling the gain of their target neurons9. Our experiments reveal high specificity in the subcellular organization of excitatory circuits.
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References
Douglas, R. J. & Martin, K. A. Mapping the matrix: the ways of neocortex. Neuron 56, 226–238 (2007)
London, M. & Hausser, M. Dendritic computation. Annu. Rev. Neurosci. 28, 503–532 (2005)
White, E. L. Specificity of cortical synaptic connectivity: emphasis on perspectives gained from quantitative electron microscopy. J. Neurocytol. 31, 195–202 (2002)
Dantzker, J. L. & Callaway, E. M. Laminar sources of synaptic input to cortical inhibitory interneurons and pyramidal neurons. Nature Neurosci. 3, 701–707 (2000)
Shepherd, G. M. G., Stepanyants, A., Bureau, I., Chklovskii, D. B. & Svoboda, K. Geometric and functional organization of cortical circuits. Nature Neurosci. 8, 782–790 (2005)
Nagel, G. et al. Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc. Natl Acad. Sci. USA 100, 13940–13945 (2003)
Petreanu, L., Huber, D., Sobczyk, A. & Svoboda, K. Channelrhodopsin-2-assisted circuit mapping of long-range callosal projections. Nature Neurosci. 10, 663–668 (2007)
Zhang, F., Wang, L. P., Boyden, E. S. & Deisseroth, K. Channelrhodopsin-2 and optical control of excitable cells. Nature Methods 3, 785–792 (2006)
Larkum, M. E., Senn, W. & Luscher, H. R. Top-down dendritic input increases the gain of layer 5 pyramidal neurons. Cereb. Cortex 14, 1059–1070 (2004)
Shu, Y., Yu, Y., Yang, J. & McCormick, D. A. Selective control of cortical axonal spikes by a slowly inactivating K+ current. Proc. Natl Acad. Sci. USA 104, 11453–11458 (2007)
Williams, S. R. & Mitchell, S. J. Direct measurement of somatic voltage clamp errors in central neurons. Nature Neurosci. 11, 790–798 (2008)
Binzegger, T., Douglas, R. J. & Martin, K. A. A quantitative map of the circuit of cat primary visual cortex. J. Neurosci. 24, 8441–8453 (2004)
Gilbert, C. D. Microcircuitry of the visual cortex. Annu. Rev. Neurosci. 6, 217–247 (1983)
Feldmeyer, D., Lubke, J. & Sakmann, B. Efficacy and connectivity of intracolumnar pairs of layer 2/3 pyramidal cells in the barrel cortex of juvenile rats. J. Physiol. (Lond.) 575, 583–602 (2006)
Lu, S. M. & Lin, R. C. S. Thalamic afferents of the rat barrel cortex: a light- and electron-microscopic study using Phaseolus vulgaris leucoagglutinin as an anterograde tracer. Somatosens. Mot. Res. 10, 1–16 (1993)
Koralek, K. A., Jensen, K. F. & Killackey, H. P. Evidence for two complementary patterns of thalamic input to the rat somatosensory cortex. Brain Res. 463, 346–351 (1988)
Veinante, P. & Deschenes, M. Single-cell study of motor cortex projections to the barrel field in rats. J. Comp. Neurol. 464, 98–103 (2003)
Lubke, J., Roth, A., Feldmeyer, D. & Sakmann, B. Morphometric analysis of the columnar innervation domain of neurons connecting layer 4 and layer 2/3 of juvenile rat barrel cortex. Cereb. Cortex 13, 1051–1063 (2003)
Hoogland, P. V., Welker, E. & Van der Loos, H. Organization of the projections from barrel cortex to thalamus in mice studied with Phaseolus vulgaris-leucoagglutinin and HRP. Exp. Brain Res. 68, 73–87 (1987)
Mainen, Z. F. & Sejnowski, T. J. Influence of dendritic structure on firing pattern in model neocortical neurons. Nature 382, 363–366 (1996)
Bureau, I., von Saint Paul, F. & Svoboda, K. Interdigitated paralemniscal and lemniscal pathways in the mouse barrel cortex. PLoS Biol. 4, e382 (2006)
Thomson, A. M. & Bannister, A. P. Interlaminar connections in the neocortex. Cereb. Cortex 13, 5–14 (2003)
Yu, C., Derdikman, D., Haidarliu, S. & Ahissar, E. Parallel thalamic pathways for whisking and touch signals in the rat. PLoS Biol. 4, e124 (2006)
Berg, R. W. & Kleinfeld, D. Vibrissa movement elicited by rhythmic electrical microstimulation to motor cortex in the aroused rat mimics exploratory whisking. J. Neurophysiol. 90, 2950–2963 (2003)
Harvey, C. D. & Svoboda, K. Locally dynamic synaptic learning rules in pyramidal neuron dendrites. Nature 450, 1195–1200 (2007)
Polsky, A., Mel, B. W. & Schiller, J. Computational subunits in thin dendrites of pyramidal cells. Nature Neurosci. 7, 621–627 (2004)
Losonczy, A. & Magee, J. C. Integrative properties of radial oblique dendrites in hippocampal CA1 pyramidal neurons. Neuron 50, 291–307 (2006)
Saito, T. & Nakatsuji, N. Efficient gene transfer into the embryonic mouse brain using in vivo electroporation. Dev. Biol. 240, 237–246 (2001)
Atasoy, D., Aponte, Y., Su, H. H. & Sternson, S. M. A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping. J. Neurosci. 28, 7025–7030 (2008)
Liao, G. Y. & Xu, B. Cre recombinase-mediated gene deletion in layer 4 of murine sensory cortical areas. Genesis 46, 289–293 (2008)
Shepherd, G. M., Pologruto, T. A. & Svoboda, K. Circuit analysis of experience-dependent plasticity in the developing rat barrel cortex. Neuron 38, 277–289 (2003)
Williams, S. R. & Stuart, G. J. Site independence of EPSP time course is mediated by dendritic I(h) in neocortical pyramidal neurons. J. Neurophysiol. 83, 3177–3182 (2000)
Nevian, T., Larkum, M. E., Polsky, A. & Schiller, J. Properties of basal dendrites of layer 5 pyramidal neurons: a direct patch-clamp recording study. Nature Neurosci. 10, 206–214 (2007)
Stepanyants, A., Hof, P. R. & Chklovskii, D. B. Geometry and structural plasticity of synaptic connectivity. Neuron 34, 275–288 (2002)
Acknowledgements
We thank A. Karpova for help with viral constructs, G. Oliver and B. Xu for the Six3Cre mouse line, D. Chklovskii, G. Shepherd and Q. Wen for comments on the manuscript, Y. Yu for the model of the dendrotoxin-sensitive potassium channel and T. O’Connor for software development.
Author Contributions L.P. and K.S. designed the experiments. L.P. performed the experiments with help from T.M. (viral injections in M1 and related recordings). S.S. provided critical reagents. L.P. and K.S. analysed the data and wrote the paper.
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This file contains This file contains Supplementary Methods, a Supplementary Discussion, Supplementary References, Supplementary Table 1 and Supplementary Figures S1-S9 with Legends Methods, a Supplementary Discussion, Supplementary References, Supplementary Table 1 and Supplementary Figures S1-S9 with Legends (PDF 3903 kb)
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Petreanu, L., Mao, T., Sternson, S. et al. The subcellular organization of neocortical excitatory connections. Nature 457, 1142–1145 (2009). https://doi.org/10.1038/nature07709
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DOI: https://doi.org/10.1038/nature07709
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