Flat-floored Air-lifted Platform: A New Method for Combining Behavior with Microscopy or Electrophysiology on Awake Freely Moving Rodents

It is widely acknowledged that the use of general anesthetics can undermine the relevance of electrophysiological or microscopical data obtained from a living animal’s brain. Moreover, the lengthy recovery from anesthesia limits the frequency of repeated recording/imaging episodes in longitudinal studies. Hence, new methods that would allow stable recordings from non-anesthetized behaving mice are expected to advance the fields of cellular and cognitive neurosciences. Existing solutions range from mere physical restraint to more sophisticated approaches, such as linear and spherical treadmills used in combination with computer-generated virtual reality. Here, a novel method is described where a head-fixed mouse can move around an air-lifted mobile homecage and explore its environment under stress-free conditions. This method allows researchers to perform behavioral tests (e.g., learning, habituation or novel object recognition) simultaneously with two-photon microscopic imaging and/or patch-clamp recordings, all combined in a single experiment. This video-article describes the use of the awake animal head fixation device (mobile homecage), demonstrates the procedures of animal habituation, and exemplifies a number of possible applications of the method.

[1]  David S. Greenberg,et al.  Population imaging of ongoing neuronal activity in the visual cortex of awake rats , 2008, Nature Neuroscience.

[2]  D. Tank,et al.  A Miniature Head-Mounted Two-Photon Microscope High-Resolution Brain Imaging in Freely Moving Animals , 2001, Neuron.

[3]  D. Tank,et al.  Intracellular dynamics of hippocampal place cells during virtual navigation , 2009, Nature.

[4]  K. Svoboda,et al.  Neural Activity in Barrel Cortex Underlying Vibrissa-Based Object Localization in Mice , 2010, Neuron.

[5]  Peter H. Barry,et al.  JPCalc, a software package for calculating liquid junction potential corrections in patch-clamp, intracellular, epithelial and bilayer measurements and for correcting junction potential measurements , 1994, Journal of Neuroscience Methods.

[6]  K. Svoboda,et al.  Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window , 2009, Nature Protocols.

[7]  John A. King,et al.  How vision and movement combine in the hippocampal place code , 2012, Proceedings of the National Academy of Sciences.

[8]  Cornelius Schwarz,et al.  Central signals rapidly switch tactile processing in rat barrel cortex during whisker movements. , 2006, Cerebral cortex.

[9]  J. Tiago Gonçalves,et al.  Internally Mediated Developmental Desynchronization of Neocortical Network Activity , 2009, The Journal of Neuroscience.

[10]  P. Golshani,et al.  Cellular mechanisms of brain-state-dependent gain modulation in visual cortex , 2013, Nature Neuroscience.

[11]  B. Sakmann,et al.  Whisker movements evoked by stimulation of single pyramidal cells in rat motor cortex , 2004, Nature.

[12]  David S. Greenberg,et al.  Visually evoked activity in cortical cells imaged in freely moving animals , 2009, Proceedings of the National Academy of Sciences.

[13]  B. Antkowiak,et al.  Neocortex is the major target of sedative concentrations of volatile anaesthetics: strong depression of firing rates and increase of GABAA receptor‐mediated inhibition , 2005, The European journal of neuroscience.

[14]  Nanxin Li,et al.  mTOR-Dependent Synapse Formation Underlies the Rapid Antidepressant Effects of NMDA Antagonists , 2010, Science.

[15]  O. Garaschuk,et al.  Targeted bulk-loading of fluorescent indicators for two-photon brain imaging in vivo , 2006, Nature Protocols.

[16]  J. G. Mcelligott,et al.  A method for restraining awake rats using head immobilization , 1993, Physiology & Behavior.

[17]  Maik C. Stüttgen,et al.  The Head-fixed Behaving Rat—Procedures and Pitfalls , 2010, Somatosensory & motor research.

[18]  Detlef H. Heck,et al.  A technique for stereotaxic recordings of neuronal activity in awake, head-restrained mice , 2009, Journal of Neuroscience Methods.

[19]  Christopher D. Harvey,et al.  Choice-specific sequences in parietal cortex during a virtual-navigation decision task , 2012, Nature.

[20]  Carlos Portera-Cailliau,et al.  A craniotomy surgery procedure for chronic brain imaging. , 2008, Journal of visualized experiments : JoVE.

[21]  D. Tank,et al.  Imaging Large-Scale Neural Activity with Cellular Resolution in Awake, Mobile Mice , 2007, Neuron.

[22]  C. Petersen,et al.  Correlating whisker behavior with membrane potential in barrel cortex of awake mice , 2006, Nature Neuroscience.

[23]  B. Sakmann,et al.  Spiking in primary somatosensory cortex during natural whisking in awake head-restrained rats is cell-type specific , 2009, Proceedings of the National Academy of Sciences.

[24]  D. Kleinfeld,et al.  Fluctuating and sensory-induced vasodynamics in rodent cortex extend arteriole capacity , 2011, Proceedings of the National Academy of Sciences.

[25]  Olav Solgaard,et al.  In vivo brain imaging using a portable 2.9 g two-photon microscope based on a microelectromechanical systems scanning mirror. , 2009, Optics letters.

[26]  M. Brecht,et al.  Behavioural report of single neuron stimulation in somatosensory cortex , 2008, Nature.

[27]  Yutaka Sakai,et al.  Reinforcing operandum: rapid and reliable learning of skilled forelimb movements by head-fixed rodents. , 2012, Journal of neurophysiology.

[28]  Roustem Khazipov,et al.  Isoflurane suppresses early cortical activity , 2013, Annals of clinical and translational neurology.

[29]  C. I. Zeeuw,et al.  Impact of conventional anesthesia on auditory brainstem responses in mice , 2004, Hearing Research.

[30]  M. Fee,et al.  Active Stabilization of Electrodes for Intracellular Recording in Awake Behaving Animals , 2000, Neuron.

[31]  D. Tank,et al.  Cellular Resolution Functional Imaging in Behaving Rats Using Voluntary Head Restraint , 2013, Neuron.

[32]  J. Poulet,et al.  Internal brain state regulates membrane potential synchrony in barrel cortex of behaving mice , 2008, Nature.

[33]  Tominaga Makoto.  European Journal of Physiologyプリューガーアーカイブの方向性 , 2016 .

[34]  Carlos Portera-Cailliau,et al.  Imaging neocortical neurons through a chronic cranial window. , 2012, Cold Spring Harbor protocols.

[35]  Lin Tian,et al.  Functional imaging of hippocampal place cells at cellular resolution during virtual navigation , 2010, Nature Neuroscience.

[36]  B. Sakmann,et al.  In vivo, low-resistance, whole-cell recordings from neurons in the anaesthetized and awake mammalian brain , 2002, Pflügers Archiv.

[37]  Maik C. Stüttgen,et al.  Two Psychophysical Channels of Whisker Deflection in Rats Align with Two Neuronal Classes of Primary Afferents , 2006, The Journal of Neuroscience.