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  1. These experiments were conducted as follows...
  2. SUBJECTS:
  3. Electrophysiological recordings were carried out in 6 adult zebra finches (Taeniopygia guttata) from the
  4. MIT zebra finch breeding facility. Birds ranged from 120-400 days post-hatch in age.
  5. Craniotomies were performed and headposts implanted on birds in order to prepare birds
  6. for head-fixation and electrophysiological recordings.
  7. HEAD FIXATION AND E-PHYS RECORDINGS:
  8. In advance of recording, birds were gradually habituated to the recording setup over the course of 3 days.
  9. Neuropixel probes (Phase 3a [Jun et al., 2017]) were used together with OpenEphys software
  10. (https://open-ephys.org/) for data acquisition, aloing with the PXIe system (National Instrument
  11. NI-PXIe-1071). Extracellular signals were amplified and filtered (0.3-10 kHz) to obtain action potential
  12. data. Craniotomies were made so that the center of the cranial aperture was centered above NCM (see
  13. surgery protocol), and the probe was lowered approximately 2000 𝜇m below the surface of the brain, until
  14. auditory responses were visible up to the 200th contact from the bottom of the probe (approximately
  15. 1500 𝜇m from the probe tip).
  16. Experiments took place on an optical table, surrounded by a home-made sound- reduction enclosure.
  17. EXPERIMENTAL STRUCTURE:
  18. This experiment involved 2 songs (consisting of 5 syllables each). The songs were played during two
  19. separate phases of the experiment. Each song was played with shuffled inter-bout intervals, consisting
  20. of 50, 100, 150, 200, 250, 300, 400, 500, 750, 1000 ms. Over the course of the experiment, each of
  21. these intervals was repeated 10 times.
  22. AUDITORY STIMULATION:
  23. Auditory stimuli were synthesized using custom MATLAB code and stimulus components from the
  24. Tchernikovskii lab zebra finch song archive [Tchernichovski et al., 2004],
  25. http://ofer.sci.ccny.cuny.edu/songs. Perturbations were introduced by inserting harmonic stacks
  26. (in the case of harmonic stack substitution), eliminating all sound (in the case of silence
  27. substitutions), or inserting spectrally matched noise (in the case of noise substitution). All
  28. substitutions were made using cosine-squared ramps to minimize edge artifacts. Spectrally-matched
  29. noise was generated by randomizing the phase of the standard, unperturbed birdsong. Sound amplitudes
  30. were matched so that substitutions (aside from silence) were as loud as the standard song piece that
  31. they replaced.
  32. Sounds were played through a PyleDriverPro speaker, with SPL of 75 dB at 20 cm. This intensity was
  33. matched to the sound level of an adult zebra finch singing at the same distance.