@article {48369, title = {Efficient method for whole-cell recording in freely moving rodents using ultraviolet-cured collar-based pipette stabilization.}, journal = {Cold Spring Harbor Protocols}, volume = {2017}, year = {2017}, month = {2017 Apr 03}, pages = {pdb.prot095810}, abstract = {

Whole-cell recording is a key technique for investigating synaptic and cellular mechanisms underlying various brain functions. However, because of its high sensitivity to mechanical disturbances, applying the whole-cell recording method to freely moving animals has been challenging. Here, we describe a technique for obtaining such recordings in freely moving, drug-free animals with a high success rate. This technique involves three major steps: obtaining a whole-cell recording from awake head-fixed animals, reliable and efficient stabilization of the pipette with respect to the animal{\textquoteright}s head using an ultraviolet (UV)-transparent collar and UV-cured adhesive, and rapid release of the animal from head fixation without loss of the recording. This technique has been successfully applied to obtain intracellular recordings from the hippocampus of freely moving rats and mice exploring a spatial environment, and should be generally applicable to other brain areas in animals engaged in a variety of natural behaviors.

}, issn = {1559-6095}, doi = {10.1101/pdb.prot095810}, author = {Lee, Doyun and Lee, Albert K} } @article {48368, title = {In vivo patch-clamp recording in awake head-fixed rodents.}, journal = {Cold Spring Harbor Protocols}, volume = {2017}, year = {2017}, month = {2017 Apr 03}, pages = {pdb.prot095802}, abstract = {

Whole-cell recording has been used to measure and manipulate a neuron{\textquoteright}s spiking and subthreshold membrane potential, allowing assessment of the cell{\textquoteright}s inputs and outputs as well as its intrinsic membrane properties. This technique has also been combined with pharmacology and optogenetics as well as morphological reconstruction to address critical questions concerning neuronal integration, plasticity, and connectivity. This protocol describes a technique for obtaining whole-cell recordings in awake head-fixed animals, allowing such questions to be investigated within the context of an intact network and natural behavioral states. First, animals are habituated to sit quietly with their heads fixed in place. Then, a whole-cell recording is obtained using an efficient, blind patching protocol. We have successfully applied this technique to rats and mice.

}, issn = {1559-6095}, doi = {10.1101/pdb.prot095802}, author = {Lee, Doyun and Lee, Albert K} } @article {48370, title = {Whole-cell recording in the awake brain.}, journal = {Cold Spring Harbor Protocols}, volume = {2017}, year = {2017}, month = {2017 Apr 03}, pages = {pdb.top087304}, abstract = {

Intracellular recording is an essential technique for investigating cellular mechanisms underlying complex brain functions. Despite the high sensitivity of the technique to mechanical disturbances, intracellular recording has been applied to awake, behaving, and even freely moving, animals. Here we summarize recent advances in these methods and their application to the measurement and manipulation of membrane potential dynamics for understanding neuronal computations in behaving animals.

}, issn = {1559-6095}, doi = {10.1101/pdb.top087304}, author = {Lee, Doyun and Lee, Albert K} } @article {45787, title = {Anesthetized- and awake-patched whole-cell recordings in freely moving rats using UV-cured collar-based electrode stabilization.}, journal = {Nature Protocols}, volume = {9}, year = {2014}, month = {2014 Nov 06}, pages = {2784-95}, abstract = {

Intracellular recording allows precise measurement and manipulation of individual neurons, but it requires stable mechanical contact between the electrode and the cell membrane, and thus it has remained challenging to perform in behaving animals. Whole-cell recordings in freely moving animals can be obtained by rigidly fixing ({\textquoteright}anchoring{\textquoteright}) the pipette electrode to the head; however, previous anchoring procedures were slow and often caused substantial pipette movement, resulting in loss of the recording or of recording quality. We describe a UV-transparent collar and UV-cured adhesive technique that rapidly (within 15 s) anchors pipettes in place with virtually no movement, thus substantially improving the reliability, yield and quality of freely moving whole-cell recordings. Recordings are first obtained from anesthetized or awake head-fixed rats. UV light cures the thin adhesive layers linking pipette to collar to head. Then, the animals are rapidly and smoothly released for recording during unrestrained behavior. The anesthetized-patched version can be completed in \~{}4-7 h (excluding histology) and the awake-patched version requires \~{}1-4 h per day for \~{}2 weeks. These advances should greatly facilitate studies of neuronal integration and plasticity in identified cells during natural behaviors.

}, issn = {1750-2799}, doi = {10.1038/nprot.2014.190}, author = {Lee, Doyun and Shtengel, Gleb and Osborne, Jason E and Lee, Albert K} } @article {5103, title = {Hippocampal place fields emerge upon single-cell manipulation of excitability during behavior.}, journal = {Science}, volume = {337}, year = {2012}, month = {2012 Aug 17}, pages = {849-53}, abstract = {

The origin of the spatial receptive fields of hippocampal place cells has not been established. A hippocampal CA1 pyramidal cell receives thousands of synaptic inputs, mostly from other spatially tuned neurons; however, how the postsynaptic neuron{\textquoteright}s cellular properties determine the response to these inputs during behavior is unknown. We discovered that, contrary to expectations from basic models of place cells and neuronal integration, a small, spatially uniform depolarization of the spatially untuned somatic membrane potential of a silent cell leads to the sudden and reversible emergence of a spatially tuned subthreshold response and place-field spiking. Such gating of inputs by postsynaptic neuronal excitability reveals a cellular mechanism for receptive field origin and may be critical for the formation of hippocampal memory representations.

}, issn = {1095-9203}, doi = {10.1126/science.1221489}, author = {Lee, Doyun and Lin, Bei-Jung and Lee, Albert K} }