Intracellular Whole-Cell Patch-Clamp Recordings of Cortical Neurons in Awake Head-Restrained Mice

  • Sylvain Crochet
Part of the Neuromethods book series (NM, volume 67)


Patch‐Clamp in Awake Mice? Membrane potential dynamics resulting from the integration of thousands of synaptic inputs and intrinsic membrane properties underlie the generation of action potential in neurons of the central nervous system. The investigation of membrane potential dynamics is, therefore, of major importance to the understanding of brain function. This level of neuronal activity can only be assessed by measuring differences of potential between the inside and the outside of a neuron, i.e., intracellular recording. In mammals, this approach has been so far mainly restricted to reduced preparations in vitro and more recently to the intact brain in anesthetized animals. Such preparations do not reproduce the complexity and the diversity of the brain activities observed in behaving animals and are, therefore, of limited interest to the understanding of complex brain processing and cognitive functions. Recently, we have developed an approach that enables intracellular recordings of cortical neurons in awake behaving mice. The mechanical stability of the brain being the main technical issue, it has been successfully overcome by (1) using “blind” whole-cell patch-clamp technique conferring higher stability in the initial phase of the recording, (2) implanting mice with light metal posts that enable painless and stable fixation of the head, (3) habituating the animal to avoid large and brisk body movements during the recording session, and (4) reducing the size of the craniotomy to minimal to prevent large brain pulsations and edema. This technique has been successfully applied to the investigation of cortical sensory processing during active sensing in the mouse whisker system and has been expanded to simultaneous double intracellular recordings or combined with other recording techniques, such as local field potentials or two-photon microscopy.

Key words

Patch clamp Intracellular recording In vivo Cortex Behaving animal 


  1. 1.
    Cragg B (1967) The density of synapses and neurones in the motor and visual areas of the cerebral cortex. J Anat 101:639–654PubMedGoogle Scholar
  2. 2.
    DeFelipe J, Farinas I (1992) The pyramidal neuron of the cerebral cortex: morphological and chemical characteristics of the synaptic inputs. Prog Neurobiol 39:563–607PubMedCrossRefGoogle Scholar
  3. 3.
    Yuste R, Tank DW (1996) Dendritic integration in mammalian neurons, a century after Cajal. Neuron 16:701–716PubMedCrossRefGoogle Scholar
  4. 4.
    Williams SR, Stuart GJ (2003) Role of dendritic synapse location in the control of action potential output. Trends Neurosci 26:147–154PubMedCrossRefGoogle Scholar
  5. 5.
    Magee JC (2000) Dendritic integration of excitatory synaptic input. Nat Rev Neurosci 1:181–190PubMedCrossRefGoogle Scholar
  6. 6.
    Reyes A, Lujan R, Rozov A et al (1998) Target-cell-specific facilitation and depression in neocortical circuits. Nat Neurosci 1:279–285PubMedCrossRefGoogle Scholar
  7. 7.
    Hubel D (1959) Single unit activity in striate cortex of unrestrained cats. J Physiol 147:226–238PubMedGoogle Scholar
  8. 8.
    Hobson JA, McCarley RW, Wyzinski P (1975) Sleep cycle oscillation: reciprocal discharge by two brainstem neuronal groups. Science 189:55–58PubMedCrossRefGoogle Scholar
  9. 9.
    Buzsaki G, Czeh G (1981) Commissural and perforant path interactions in the rat hippocampus. Field potentials and unitary activity. Exp Brain Res 43:429–438PubMedCrossRefGoogle Scholar
  10. 10.
    Romo R, Salinas E (2001) Touch and go: decision-making mechanisms in somatosensation. Annu Rev Neurosci 24:107–137PubMedCrossRefGoogle Scholar
  11. 11.
    Wiest MC, Bentley N, Nicolelis MA (2005) Heterogeneous integration of bilateral whisker signals by neurons in primary somatosensory cortex of awake rats. J Neurophysiol 93:2966–2973PubMedCrossRefGoogle Scholar
  12. 12.
    Huetz C, Philibert B, Edeline JM (2009) A spike-timing code for discriminating conspecific vocalizations in the thalamocortical system of anesthetized and awake guinea pigs. J Neurosci 29:334–350PubMedCrossRefGoogle Scholar
  13. 13.
    Ecker AS, Berens P, Keliris GA et al (2010) Decorrelated neuronal firing in cortical microcircuits. Science 327:584–587PubMedCrossRefGoogle Scholar
  14. 14.
    von Heimendahl M, Itskov PM, Arabzadeh E et al (2007) Neuronal activity in rat barrel cortex underlying texture discrimination. PLoS Biol 5:e305CrossRefGoogle Scholar
  15. 15.
    Krupa DJ, Wiest MC, Shuler MG et al (2004) Layer-specific somatosensory cortical activation during active tactile discrimination. Science 304:1989–1992PubMedCrossRefGoogle Scholar
  16. 16.
    Sakai K, Crochet S (2001) Differentiation of presumed serotonergic dorsal raphe neurons in relation to behavior and wake-sleep states. Neuroscience 104:1141–1155PubMedCrossRefGoogle Scholar
  17. 17.
    Csicsvari J, Henze DA, Jamieson B et al (2003) Massively parallel recording of unit and local field potentials with silicon-based electrodes. J Neurophysiol 90:1314–1323PubMedCrossRefGoogle Scholar
  18. 18.
    Pastalkova E, Itskov V, Amarasingham A et al (2008) Internally generated cell assembly sequences in the rat hippocampus. Science 321:1322–1327PubMedCrossRefGoogle Scholar
  19. 19.
    Peyrache A, Khamassi M, Benchenane K et al (2009) Replay of rule-learning related neural patterns in the prefrontal cortex during sleep. Nat Neurosci 12:919–926PubMedCrossRefGoogle Scholar
  20. 20.
    Lin SC, Nicolelis MA (2008) Neuronal ensemble bursting in the basal forebrain encodes salience irrespective of valence. Neuron 59:138–149PubMedCrossRefGoogle Scholar
  21. 21.
    Timofeev I, Grenier F, Steriade M (2001) Disfacilitation and active inhibition in the neocortex during the natural sleep-wake cycle: an intracellular study. Proc Natl Acad Sci USA 98:1924–1929PubMedCrossRefGoogle Scholar
  22. 22.
    Mahon S, Vautrelle N, Pezard L et al (2006) Distinct patterns of striatal medium spiny neuron activity during the natural sleep-wake cycle. J Neurosci 26:12587–12595PubMedCrossRefGoogle Scholar
  23. 23.
    Okun M, Naim A, Lampl I (2010) The subthreshold relation between cortical local field potential and neuronal firing unveiled by intracellular recordings in awake rats. J Neurosci 30:4440–4448PubMedCrossRefGoogle Scholar
  24. 24.
    Petersen CC, Hahn TT, Mehta M et al (2003) Interaction of sensory responses with spontaneous depolarization in layer 2/3 barrel cortex. Proc Natl Acad Sci USA 100:13638–13643PubMedCrossRefGoogle Scholar
  25. 25.
    Gentet LJ, Avermann M, Matyas F et al (2010) Membrane potential dynamics of GABAergic neurons in the barrel cortex of behaving mice. Neuron 65:422–435PubMedCrossRefGoogle Scholar
  26. 26.
    Edeline JM, Dutrieux G, Manunta Y et al (2001) Diversity of receptive field changes in auditory cortex during natural sleep. Eur J Neurosci 14:1865–1880PubMedCrossRefGoogle Scholar
  27. 27.
    Castro-Alamancos MA (2004) Absence of rapid sensory adaptation in neocortex during information processing states. Neuron 41:455–464PubMedCrossRefGoogle Scholar
  28. 28.
    Ferezou I, Bolea S, Petersen CC (2006) Visualizing the cortical representation of whisker touch: voltage-sensitive dye imaging in freely moving mice. Neuron 50:617–629PubMedCrossRefGoogle Scholar
  29. 29.
    Petersen CC (2007) The functional organization of the barrel cortex. Neuron 56:339–355PubMedCrossRefGoogle Scholar
  30. 30.
    Brecht M (2007) Barrel cortex and whisker-mediated behaviors. Curr Opin Neurobiol 17:408–416PubMedCrossRefGoogle Scholar
  31. 31.
    Carvell GE, Simons DJ (1990) Biometric analyses of vibrissal tactile discrimination in the rat. J Neurosci 10:2638–2648PubMedGoogle Scholar
  32. 32.
    Prigg T, Goldreich D, Carvell GE et al (2002) Texture discrimination and unit recordings in the rat whisker/barrel system. Physiol Behav 77:671–675PubMedCrossRefGoogle Scholar
  33. 33.
    Knutsen PM, Pietr M, Ahissar E (2006) Haptic object localization in the vibrissal system: behavior and performance. J Neurosci 26:8451–8464PubMedCrossRefGoogle Scholar
  34. 34.
    O’Connor DH, Clack NG, Huber D et al (2010) Vibrissa-based object localization in head-fixed mice. J Neurosci 30:1947–1967PubMedCrossRefGoogle Scholar
  35. 35.
    Crochet S, Petersen CC (2006) Correlating whisker behavior with membrane potential in barrel cortex of awake mice. Nat Neurosci 9:608–610PubMedCrossRefGoogle Scholar
  36. 36.
    Poulet JF, Petersen CC (2008) Internal brain state regulates membrane potential synchrony in barrel cortex of behaving mice. Nature 454:881–885PubMedCrossRefGoogle Scholar
  37. 37.
    Harvey CD, Collman F, Dombeck DA et al (2009) Intracellular dynamics of hippocampal place cells during virtual navigation. Nature 461:941–946PubMedCrossRefGoogle Scholar
  38. 38.
    Lee AK, Manns ID, Sakmann B et al (2006) Whole-cell recordings in freely moving rats. Neuron 51:399–407PubMedCrossRefGoogle Scholar
  39. 39.
    Epsztein J, Lee AK, Chorev E et al (2010) Impact of spikelets on hippocampal CA1 pyramidal cell activity during spatial exploration. Science 327:474–477PubMedCrossRefGoogle Scholar
  40. 40.
    Souliere F, Urbain N, Gervasoni D et al (2000) Single-unit and polygraphic recordings associated with systemic or local pharmacology: a multi-purpose stereotaxic approach for the awake, anaesthetic-free, and head-restrained rat. J Neurosci Res 61: 88–100PubMedCrossRefGoogle Scholar
  41. 41.
    Takahashi K, Lin JS, Sakai K (2006) Neuronal activity of histaminergic tuberomammillary neurons during wake-sleep states in the mouse. J Neurosci 26:10292–10298PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Sylvain Crochet
    • 1
    • 2
  1. 1.INSERM/UCBL-U628, Integrated Physiology of Brain Arousal SystemsClaude Bernard University Lyon 1LyonFrance
  2. 2.Laboratory of Sensory Processing, Brain Mind InstituteÉcole Polytechnique Fédérale de Lausanne (EPFL)LausanneSwitzerland

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