An affordable solution for investigating zebra finch intracranial electroencephalography (iEEG) signals.

The zebra finch is a well-studied animal model for investigating the neural mechanisms of vocal learning, and electrophysiology is the primary technique for understanding their song system. Most of the studies on zebra finches have focused on intracerebral recordings. However, these methods are only affordable for limited laboratories. Recently, different open-source hardware for acquiring electroencephalography (EEG) signals has been developed. It's unclear whether these solutions suit zebra finch studies as they have not been evaluated. Electrocorticography signals can provide a preliminary guide for more in-depth inquiries and also aid in understanding the global behavior of the bird's brain, as opposed to the more common localized approach. We present a detailed protocol for acquiring intracranial electroencephalography (iEEG) data from zebra finches using the OpenBCI Cyton board, an open-source device. We implemented stainless steel electrodes on the brain's surface and recorded the brain signals from two recording sites above two auditory-responsive nuclei. To validate our method, we ran two different experiments. In the first experiment, we recorded neural activity under various concentrations of isoflurane and extracted the suppression duration to measure anesthesia depth. In the second experiment, we head-fixed the birds and, under light anesthesia, presented them with various auditory stimuli to evaluate event-related potentials (ERP) and generate spectrograms. The results showed a significant increase in suppression duration with deeper anesthesia, and the ERP and spectrogram responses to auditory stimuli differed accordingly. These findings indicate that using our methodology, one can successfully collect iEEG signals from zebra finches. These findings pave the way for future studies to use iEEG to investigate bird cognition in a more affordable way.