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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 3 distinct songs presented in 2 distinct epochs. The songs each consisted
  19. of 5 syllables, and were repeated in bouts of 5 repetitions each. During the first epoch, the bouts
  20. are all separated by a standard inter-bout interval (called 𝜏0). This was 100ms for the first song,
  21. 1000ms for the second song, and 10000ms for the third song. This standard inter-bout interval was
  22. presented for 25 bouts. During the second epoch, inter-bout intervals were randomly chosen to be
  23. between 0.5 and 1.5 times 𝜏0, uniformly distributed. The experiment was performed so that each both
  24. epochs were completed with the first song before the second song was played. Then, both epochs were
  25. completed with the second song before the third song was played.
  26. AUDITORY STIMULATION:
  27. Auditory stimuli were synthesized using custom MATLAB code and stimulus components from the
  28. Tchernikovskii lab zebra finch song archive [Tchernichovski et al., 2004],
  29. http://ofer.sci.ccny.cuny.edu/songs. Perturbations were introduced by inserting harmonic stacks
  30. (in the case of harmonic stack substitution), eliminating all sound (in the case of silence
  31. substitutions), or inserting spectrally matched noise (in the case of noise substitution). All
  32. substitutions were made using cosine-squared ramps to minimize edge artifacts. Spectrally-matched
  33. noise was generated by randomizing the phase of the standard, unperturbed birdsong. Sound amplitudes
  34. were matched so that substitutions (aside from silence) were as loud as the standard song piece that
  35. they replaced.
  36. Sounds were played through a PyleDriverPro speaker, with SPL of 75 dB at 20 cm. This intensity was
  37. matched to the sound level of an adult zebra finch singing at the same distance.