Abstract
Humans and animals use predictions to optimize their behavior, however, the underlying neuronal implementation remains elusive. We address this using omitted sounds on the macro- and microscale of the auditory cortex of female, normal hearing mice using high-speed imaging. Neuronal responses to the omission of expected sounds were time-locked to expected stimulus onset, localized to layer 1-4 of higher auditory area (Temporal Association Area, TeA), and continued to rise until the following stimulus. The omission responses differed from offset and deviant responses in their temporal shape, size and spatial localization. Omissions and sequence statistics correlated with behavioral changes by timed pupil dilation and rapid facial motions. While stimulus responses showed partial entrainment, omission responses maintained a distinct, unentrained shape. The localized omission response in TeA is consistent with a hierarchical organization of predictive processing. However, the continued rise suggests an integrated, absolute prediction error, instead of a direct representation of prediction or prediction error, which would terminate with the omission.
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Data availability
The neural and video data generated in this study, as well as all files necessary for the creation of all figures, have been deposited in the open-access Radboud Repository (doi: 10.34973/e6ph-q277) in a compressed (i.e., preprocessed) format. Source Data for each figure is organized under Repository/Source Data/FigureName/[..]. For more details on data structure, refer to the README.txt file in the base directory of the repository. The raw neural and video datasets were too large for online storage, but will be made available upon request to the corresponding author.
Code availability
All code used for stimulus generation, data acquisition, preprocessing, data analysis, and figure plotting is also made available via the same Radboud Repository as the Data, fixed from the time of publication (doi: 10.34973/e6ph-q277). Inside /Code/, we provide code for the creation of each figure separately under the subfolder /Figures/. Subfolders inside /Figures/ are structured per Figure, according to the format ‘Figure_X_ShortTitle’. For more details on code structure, refer to the README.txt file in the base directory of the repository.
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Acknowledgements
We would like to thank Jeroen Bos and the support staff of INSS for their assistance during the development of the experimental setup. We would like to thank Floris de Lange and Uta Noppeney for helpful discussions and feedback on earlier versions of the manuscript. We would like to thank Daniel Polley, Ross Williamson, Bruno Pichler, Yannick Goullam-Houssen, Brice Batthelier, and Rémi Proville for initial discussions on the experimental setup design and Roberta Müller for assistance with the illustrations of experimental setups. Further, we would like to thank Gesa Berretz, Karol Przewrocki, and Artoghrul Alishbayli for insightful discussions. Bernhard Englitz acknowledges funding from a VIDI grant (016.VIDI.189.052) and Zhongnan Cai from an internal grant at the Donders Center for Neuroscience. Bernhard Englitz also acknowledges valuable discussions at the Kavli Conference on Statistical Learning in the Brain at UCSB, supported by NSF Grant No. PHY-1748958 and the Gordon and Betty Moore Foundation Grant No. 2919.02 to the Kavli Institute for Theoretical Physics (KITP). Francesco P. Battaglia acknowledges funding from the European Research Council (ERC) Advanced Grant “REPLAY-DMN” (grant agreement no. 833964).
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Bernhard Englitz acquired funding, conceived and implemented the experiments, managed and supervised the project, and wrote the manuscript. Janek Peters developed, managed, and performed the surgeries and imaging experiments, wrote analyses and generalized analysis tools, co-developed and managed the code pipeline, supervised students, produced figures, and co-wrote the manuscript. Zhongnan Cai co-developed the analysis pipeline, in particular, all aspects of 2P data processing and visualization, and contributed significantly to manuscript writing. Maxime van Veghel developed pilot analyses for the omission control paradigms. Andreas Knoben implemented facial and pupil tracking and piloted analysis. Maikel Simon acquired imaging data. Samuel Arends assisted with facial motion tracking and pipeline improvements. Francesco P. Battaglia acquired funding for and managed the acquisition as well as the assembly of the 2p microscope. All authors read and approved the final manuscript.
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Peters, J., Cai, Z., van Veghel, M. et al. The representation of omitted sounds in the mouse auditory cortex. Nat Commun (2026). https://doi.org/10.1038/s41467-026-68847-w
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DOI: https://doi.org/10.1038/s41467-026-68847-w
