Anesthetized- and awake-patched whole-cell recordings in freely moving rats using UV-cured collar-based electrode stabilization.
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 ('anchoring') 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.
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, Virginia, USA.
Doyun
D
Lee
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, Virginia, USA.
Gleb
G
Shtengel
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, Virginia, USA.
Jason E
JE
Osborne
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, Virginia, USA.
Albert K
AK
Lee
D000758:Anesthesia / Q000379:methods; D000818:Animals; D001522:Behavior, Animal; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D006624:Hippocampus / Q000166:cytology; D009043:Motor Activity; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D017208:Rats, Wistar; D015203:Reproducibility of Results; D014466:Ultraviolet Rays; D014851:Wakefulness