Behaviour-dependent recruitment of long-range projection neurons in somatosensory cortex

Journal name:
Nature
Volume:
499,
Pages:
336–340
Date published:
DOI:
doi:10.1038/nature12236
Received
Accepted
Published online

In the mammalian neocortex, segregated processing streams are thought to be important for forming sensory representations of the environment1, 2, but how local information in primary sensory cortex is transmitted to other distant cortical areas during behaviour is unclear. Here we show task-dependent activation of distinct, largely non-overlapping long-range projection neurons in the whisker region of primary somatosensory cortex (S1) in awake, behaving mice. Using two-photon calcium imaging, we monitored neuronal activity in anatomically identified S1 neurons projecting to secondary somatosensory (S2) or primary motor (M1) cortex in mice using their whiskers to perform a texture-discrimination task or a task that required them to detect the presence of an object at a certain location. Whisking-related cells were found among S2-projecting (S2P) but not M1-projecting (M1P) neurons. A higher fraction of S2P than M1P neurons showed touch-related responses during texture discrimination, whereas a higher fraction of M1P than S2P neurons showed touch-related responses during the detection task. In both tasks, S2P and M1P neurons could discriminate similarly between trials producing different behavioural decisions. However, in trials producing the same decision, S2P neurons performed better at discriminating texture, whereas M1P neurons were better at discriminating location. Sensory stimulus features alone were not sufficient to elicit these differences, suggesting that selective transmission of S1 information to S2 and M1 is driven by behaviour.

At a glance

Figures

  1. In vivo calcium imaging of long-range projection neurons in S1[thinsp]during texture discrimination.
    Figure 1: In vivo calcium imaging of long-range projection neurons in S1during texture discrimination.

    a, Retrograde labelling of S2Pand M1Pneurons inS1 by AAV6-Cre and CTB-Alexa647 injection, respectively. AAV6-YC-Nano140 was injected into S1 for calcium imaging. b, Top, in vivo image of L2/3 neurons in S1 expressing YC-Nano140 (white). AAV6-Cre-infected S2P neurons express tdTomato (red). Middle and bottom (lower magnification), post-hoc identification of in vivo imaged neurons with CTB-Alexa647-labelled M1Pneurons (blue) and immunostained GABA-positive neurons (green). Scale bar, 20µm. c, Setup for two-photon (2P) imaging of S1 neurons during head-fixed texture discrimination. d, Trial structure for go/no-go texture discrimination task. CR, correct rejection; FA, false alarm. e, Calcium transients (black) from example cells and the neuropil (NP) across nine trials, and the average trace across all trials. Envelope of whisking amplitude (green) and periods of touch (orange area) are also shown. UNL, unlabelled. f, Cross-correlation analysis of calcium signals with whisking amplitude (green) and touch (orange) across different time lags for ‘whisking’ (cell 1) and ‘touch’ (cell 2) in e. Shaded trace indicates 95% confidence interval from bootstrap test. Grey area indicates lag window for classification. g, Average calcium trace across all whisking (green), touch (orange), and unclassified (grey) neurons shown with average whisking amplitude and touch vectors (dotted lines). h, Distribution of classified cells across subtypes. Error bars, s.d. from bootstrap test. A permutation test of shuffled labels are shown. Solid horizontal lines, means; dashed horizontal lines, 95% confidence intervals (n = 231 active neurons). *P<0.05.

  2. Single-neuron discrimination analysis of decision or texture in S1 projection neurons.
    Figure 2: Single-neuron discrimination analysis of decision or texture in S1 projection neurons.

    a, Single-trial responses of example S2P, M1P or UNL touch neurons to trial type or texture aligned to first touch (dotted line). b, Average calcium transient of neurons in a according to trial type or texture. Shaded areas, s.e.m. c, Fraction of trials in which individual cells correctly discriminated between decision (hit versus correct rejection) or between non-target textures from ROC analysis. Grey line indicates the 95th percentile of distribution from a permutation test of decision or texture labels. Neurons are ranked according to the fraction of trials that were correctly discriminated. Neurons above this line can discriminate above chance. d, Fraction of active cells discriminating trial type or texture above chance. e, Performance of neurons discriminating above chance. Circles indicate individual neurons shaded according to their behaviour classification. Grey lines indicate 95th percentile of distribution from a permutation test of decision or texture labels. Error bars, s.d. from a permutation test (d), s.e.m. (e). n = 231 active neurons; *P<0.05.

  3. Activity of S1 projection neurons during object localization.
    Figure 3: Activity of S1 projection neurons during object localization.

    a, Positions of poles during the object-localization task. b, Calcium transients (black) of example cells and the neuropil (NP) across four trials, along with the average traces across all trials. Whisking amplitude (green) and periods of pole touch (orange area) are also shown. c, Average calcium trace across all whisking (green), touch (orange) and unclassified (grey) neurons shown with average whisking amplitude and touch vector (dotted lines). d, Distribution of classified cells across subtypes. A permutation test of shuffled labels is shown. Solid line, mean; dashed line, 95% confidence interval. e, Fraction of cells that discriminate decision or pole position above chance, from a ROC analysis. f, Performance of neurons discriminating above chance. Circles indicate individual neurons shaded according to behaviour classification. Grey line indicates the 95th percentile of distribution from a permutation test of decision or position labels. Error bars, s.d. from bootstrap test (d), s.d. from permutation test (e), s.e.m. (f). n = 180 active neurons; *P<0.05, **P<0.01.

  4. Sensory stimuli are not sufficient to produce task-related differences.
    Figure 4: Sensory stimuli are not sufficient to produce task-related differences.

    a, Whisker kinematic differences during texture discrimination versus object localization for high-acceleration-velocity ‘slip’ events, maximum absolute curvature change (max |Δκ|), mean touch angle (0° = orthogonal to anterior–posterior axis) over first second of touch. b, Cross correlation of calcium signals with whisker kinematic features for touch during texture discrimination (top panel) or object localization (bottom panel). c, Fraction of touch neurons identified by cross-correlation analysis from active neurons in naive animals. A permutation test of shuffled labels is shown. d, Cross correlation of calcium activity to sandpaper versus pole touch in naive animals. Inset, population bias index (see Methods) determined from the R value. e, Fraction of cells discriminating above chance for target versus non-target stimuli or between non-target stimuli, from a ROC analysis in naive animals. Error bars, s.e.m. (a), s.d. from a bootstrap test (c), s.d. from a permutation test (c). n = 1,574 trials (a), n = 98touch neurons (b); n = 207 active neurons (ce); *P<0.05, **P<0.02.

Videos

  1. Hit trial during texture discrimination
    Video 1: 'Hit trial' during texture discrimination
    Realtime video of hit trial from head-fixed mouse performing texture discrimination task during two-photon imaging. Video was acquired using an infrared CCD camera under infrared LED illumination (for high-speed whisker tracking) with additional illumination provided by two-photon excitation laser source.
  2. Correct rejection trial during texture discrimination
    Video 2: 'Correct rejection' trial during texture discrimination
    Realtime video of correct rejection trial from head-fixed mouse performing texture discrimination task during two-photon imaging. Video was acquired using an infrared CCD camera under infrared LED illumination (for high-speed whisker tracking) with additional illumination provided by two-photon excitation laser source.
  3. Neuronal activity and whisking behavior during texture discrimination
    Video 3: Neuronal activity and whisking behavior during texture discrimination
    Realtime playback of high-speed video of whisking and touch (top left panel) along with two-photon imaging of calcium activity in S1 (bottom panel) from a “correct rejection” trial. Top right panel shows measured whisker angle, whisking amplitude, and period of texture touch (orange region) along with calcium traces of active cells. Indicated cells 1-5 were behaviorally classified as touch cells with cells 1, 3-5 identified as S2P neurons and cell 2 identified as an UNL neuron.

References

  1. Diamond, M. E., von Heimendahl, M., Knutsen, P. M., Kleinfeld, D. & Ahissar, E. 'Where' and 'what' in the whisker sensorimotor system. Nature Rev. Neurosci. 9, 601612 (2008)
  2. Felleman, D. J. & Van Essen, D. C. Distributed hierarchical processing in the primate cerebral cortex. Cereb. Cortex 1, 147 (1991)
  3. Aronoff, R. et al. Long-range connectivity of mouse primary somatosensory barrel cortex. Eur. J. Neurosci. 31, 22212233 (2010)
  4. Mao, T. et al. Long-range neuronal circuits underlying the interaction between sensory and motor cortex. Neuron 72, 111123 (2011)
  5. Sato, T. R. & Svoboda, K. The functional properties of barrel cortex neurons projecting to the primary motor cortex. J. Neurosci. 30, 42564260 (2010)
  6. Chakrabarti, S. & Alloway, K. D. Differential origin of projections from SI barrel cortex to the whisker representations in SII and MI. J. Comp. Neurol. 498, 624636 (2006)
  7. Madisen, L. et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nature Neurosci. 13, 133140 (2010)
  8. Horikawa, K. et al. Spontaneous network activity visualized by ultrasensitive Ca2+ indicators, yellow Cameleon-Nano. Nature Methods 7, 729732 (2010)
  9. Carvell, G. E. & Simons, D. J. Biometric analyses of vibrissal tactile discrimination in the rat. J. Neurosci. 10, 26382648 (1990)
  10. Guic-Robles, E., Jenkins, W. M. & Bravo, H. Vibrissal roughness discrimination is barrel cortex-dependent. Behav. Brain Res. 48, 145152 (1992)
  11. Jadhav, S. P., Wolfe, J. & Feldman, D. E. Sparse temporal coding of elementary tactile features during active whisker sensation. Nature Neurosci. 12, 792800 (2009)
  12. von Heimendahl, M., Itskov, P. M., Arabzadeh, E. & Diamond, M. E. Neuronal activity in rat barrel cortex underlying texture discrimination. PLoS Biol. 5, e305 (2007)
  13. Crochet, S. & Petersen, C. C. Correlating whisker behavior with membrane potential in barrel cortex of awake mice. Nature Neurosci. 9, 608610 (2006)
  14. Fee, M. S., Mitra, P. P. & Kleinfeld, D. Central versus peripheral determinants of patterned spike activity in rat vibrissa cortex during whisking. J. Neurophysiol. 78, 11441149 (1997)
  15. Curtis, J. C. & Kleinfeld, D. Phase-to-rate transformations encode touch in cortical neurons of a scanning sensorimotor system. Nature Neurosci. 12, 492501 (2009)
  16. O'Connor, D. H., Peron, S. P., Huber, D. & Svoboda, K. Neural activity in barrel cortex underlying vibrissa-based object localization in mice. Neuron 67, 10481061 (2010)
  17. de Kock, C. P., Bruno, R. M., Spors, H. & Sakmann, B. Layer- and cell-type-specific suprathreshold stimulus representation in rat primary somatosensory cortex. J. Physiol. (Lond.) 581, 139154 (2007)
  18. Petreanu, L. et al. Activity in motor-sensory projections reveals distributed coding in somatosensation. Nature 489, 299303 (2012)
  19. Xu, N. L. et al. Nonlinear dendritic integration of sensory and motor input during an active sensing task. Nature 492, 247251 (2012)
  20. Hill, D. N., Curtis, J. C., Moore, J. D. & Kleinfeld, D. Primary motor cortex reports efferent control of vibrissa motion on multiple timescales. Neuron 72, 344356 (2011)
  21. Green, D. M. & Swets, J. A. Signal Detection Theory and Psychophysics (Wiley, 1966)
  22. O'Connor, D. H. et al. Vibrissa-based object localization in head-fixed mice. J. Neurosci. 30, 19471967 (2010)
  23. Huber, D. et al. Multiple dynamic representations in the motor cortex during sensorimotor learning. Nature 484, 473478 (2012)
  24. Mehta, S. B., Whitmer, D., Figueroa, R., Williams, B. A. & Kleinfeld, D. Active spatial perception in the vibrissa scanning sensorimotor system. PLoS Biol. 5, e15 (2007)
  25. Melzer, P., Champney, G. C., Maguire, M. J. & Ebner, F. F. Rate code and temporal code for frequency of whisker stimulation in rat primary and secondary somatic sensory cortex. Exp. Brain Res. 172, 370386 (2006)
  26. Alloway, K. D. Information processing streams in rodent barrel cortex: the differential functions of barrel and septal circuits. Cereb. Cortex 18, 979989 (2008)
  27. Guic, E., Carrasco, X., Rodriguez, E., Robles, I. & Merzenich, M. M. Plasticity in primary somatosensory cortex resulting from environmentally enriched stimulation and sensory discrimination training. Biol. Res. 41, 425437 (2008)
  28. Wiest, M. C., Thomson, E., Pantoja, J. & Nicolelis, M. A. Changes in S1 neural responses during tactile discrimination learning. J. Neurophysiol. 104, 300312 (2010)
  29. Krupa, D. J., Wiest, M. C., Shuler, M. G., Laubach, M. & Nicolelis, M. A. Layer-specific somatosensory cortical activation during active tactile discrimination. Science 304, 19891992 (2004)
  30. Gilbert, C. D. & Sigman, M. Brain states: top-down influences in sensory processing. Neuron 54, 677696 (2007)
  31. Gradinaru, V. et al. Molecular and cellular approaches for diversifying and extending optogenetics. Cell 141, 154165 (2010)
  32. Lütcke, H. et al. Optical recording of neuronal activity with a genetically-encoded calcium indicator in anesthetized and freely moving mice. Front Neural Circuits 4, 9 (2010)
  33. Margolis, D. J. et al. Reorganization of cortical population activity imaged throughout long-term sensory deprivation. Nature Neurosci. 15, 15391546 (2012)
  34. Langer, D. et al. HelioScan: A software framework for controlling in vivo microscopy setups with high hardware flexibility, functional diversity and extendibility. J. Neurosci. Methods 215, 3852 (2013)
  35. Knutsen, P. M., Derdikman, D. & Ahissar, E. Tracking whisker and head movements in unrestrained behaving rodents. J. Neurophysiol. 93, 22942301 (2004)
  36. Conte, W. L., Kamishina, H. & Reep, R. L. Multiple neuroanatomical tract-tracing using fluorescent Alexa Fluor conjugates of cholera toxin subunit B in rats. Nature Protocols 4, 11571166 (2009)
  37. Dombeck, D. A., Khabbaz, A. N., Collman, F., Adelman, T. L. & Tank, D. W. Imaging large-scale neural activity with cellular resolution in awake, mobile mice. Neuron 56, 4357 (2007)
  38. de Kock, C. P. & Sakmann, B. Spiking in primary somatosensory cortex during natural whisking in awake head-restrained rats is cell-type specific. Proc. Natl Acad. Sci. USA 106, 1644616450 (2009)
  39. Birdwell, J. A. et al. Biomechanical models for radial distance determination by the rat vibrissal system. J. Neurophysiol. 98, 24392455 (2007)

Download references

Author information

Affiliations

  1. Brain Research Institute, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland

    • Jerry L. Chen,
    • Stefano Carta,
    • Joana Soldado-Magraner &
    • Fritjof Helmchen
  2. Neuroscience Center Zurich, University of Zurich/ETH Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland

    • Stefano Carta &
    • Fritjof Helmchen
  3. Institute of Neuroinformatics, University of Zurich/ETH Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland

    • Joana Soldado-Magraner
  4. Brain Mind Institute, Ecole Polytechnique Fédérale de Lausanne (EPFL), EPFL SV BMI LEN, Station 19, CH-1015 Lausanne, Switzerland

    • Bernard L. Schneider

Contributions

J.L.C. and F.H. designed the study. J.L.C. carried out experiments. J.L.C., S.C., J.S.M and F.H. performed data analysis. S.C. carried out experiments and data analysis characterizing YC-Nano140. B.L.S. contributed viral reagents. J.L.C. and F.H. wrote the paper.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to:

Author details

Supplementary information

Video

  1. Video 1: 'Hit trial' during texture discrimination (1,161 KB, Download)
    Realtime video of hit trial from head-fixed mouse performing texture discrimination task during two-photon imaging. Video was acquired using an infrared CCD camera under infrared LED illumination (for high-speed whisker tracking) with additional illumination provided by two-photon excitation laser source.
  2. Video 2: 'Correct rejection' trial during texture discrimination (1,256 KB, Download)
    Realtime video of correct rejection trial from head-fixed mouse performing texture discrimination task during two-photon imaging. Video was acquired using an infrared CCD camera under infrared LED illumination (for high-speed whisker tracking) with additional illumination provided by two-photon excitation laser source.
  3. Video 3: Neuronal activity and whisking behavior during texture discrimination (796 KB, Download)
    Realtime playback of high-speed video of whisking and touch (top left panel) along with two-photon imaging of calcium activity in S1 (bottom panel) from a “correct rejection” trial. Top right panel shows measured whisker angle, whisking amplitude, and period of texture touch (orange region) along with calcium traces of active cells. Indicated cells 1-5 were behaviorally classified as touch cells with cells 1, 3-5 identified as S2P neurons and cell 2 identified as an UNL neuron.

PDF files

  1. Supplementary Information (3.2 MB)

    This file contains Supplementary Figures 1-14, Supplementary Methods and Supplementary Table 1.

Additional data