Abstract
The retrosplenial cortex (RSC) integrates sensory and mnemonic information to support spatial orientation and navigation, yet how visuospatial processing differs across its subregions remains unclear. Here, we combined cellular imaging in navigating mice with brain-wide anatomical input tracing to characterize how multimodal sensory and positional signals are integrated along the anterior–posterior axis of dorsal RSC. We identified consistent differences between anterior and posterior subregions in both functional response properties and long-range connectivity. Anterior RSC neurons displayed sharper and more reliable position tuning during tactile-cued navigation and preferential sensitivity to fast, low–spatial-frequency visual motion. In contrast, posterior RSC neurons showed broader position selectivity, stronger responses to slow, high–spatial-frequency visual patterns, and enhanced tuning in visually immersive virtual environments. Consistent with these differences, anterior RSC received denser projections from motor, somatosensory, and parietal areas, whereas posterior RSC received stronger input from primary and posteromedial visual cortices. Together, these findings identify an anterior–posterior functional gradient in RSC, with subregions differing in how they integrate sensory and positional signals during navigation.
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Data availability
All two-photon imaging datasets (Suite2p-processed) and retrograde-labeling data (CSV format) have been deposited in Zenodo and are publicly available (https://doi.org/10.5281/zenodo.17639283). Any additional data supporting the findings of this study are available from the corresponding author upon request.
Code availability
All custom code used in this study is openly available on GitHub (https://github.com/ytsimon2004/rscvp) and archived in Zenodo (https://doi.org/10.5281/zenodo.18234118). The public repository includes detailed documentation and Google Colab notebooks to facilitate streamlined data download and full reproduction of the analyses. Further information required to reproduce or reanalyze the results is available from the corresponding author upon request.
References
Auger, S. D., Mullally, S. L. & Maguire, E. A. Retrosplenial cortex codes for permanent landmarks. PLoS One 7, e43620 (2012).
Vann, S. D., Aggleton, J. P. & Maguire, E. A. What does the retrosplenial cortex do? Nat. Rev. Neurosci. 10, 792–802 (2009).
Alexander, A. S., Place, R., Starrett, M. J., Chrastil, E. R. & Nitz, D. A. Rethinking retrosplenial cortex: perspectives and predictions. Neuron 111, 150–175 (2023).
Saleem, A. B. & Busse, L. Interactions between rodent visual and spatial systems during navigation. Nat. Rev. Neurosci. 24, 487–501 (2023).
Epstein, R. A., Patai, E. Z., Julian, J. B. & Spiers, H. J. The cognitive map in humans: spatial navigation and beyond. Nat. Neurosci. 20, 1504–1513 (2017).
Brennan, E. K. et al. Thalamus and claustrum control parallel layer 1 circuits in retrosplenial cortex. eLife 10, e62207 (2021).
Yamawaki, N., Radulovic, J. & Shepherd, G. M. G. A corticocortical circuit directly links retrosplenial cortex to M2 in the mouse. J. Neurosci. 36, 9365–9374 (2016).
Morimoto, M. M., Uchishiba, E. & Saleem, A. B. Organization of feedback projections to mouse primary visual cortex. iScience 24, 102450 (2021).
Haugland, K. G., Sugar, J. & Witter, M. P. Development and topographical organization of projections from the hippocampus and parahippocampus to the retrosplenial cortex. Eur. J. Neurosci. 50, 1799–1819 (2019).
Miller, A. P., Vedder, L. C., Law, M. L. & Smith, D. M. Cues, context, and long-term memory: the role of the retrosplenial cortex in spatial cognition. Front. Hum. Neurosci. 8, 586 (2014).
Vedder, L. C., Miller, A. M. P., Harrison, M. B. & Smith, D. M. Retrosplenial cortical neurons encode navigational cues, trajectories and reward locations during goal directed navigation. Cereb. Cortex 27, 3713–3723 (2017).
Sun, W. et al. Context value updating and multidimensional neuronal encoding in the retrosplenial cortex. Nat. Commun. 12, 6045 (2021).
Hattori, R. & Komiyama, T. Context-dependent persistency as a coding mechanism for robust and widely distributed value coding. Neuron 110, 502–515 (2022).
Alexander, A. S. & Nitz, D. A. Retrosplenial cortex maps the conjunction of internal and external spaces. Nat. Neurosci. 18, 1143–1151 (2015).
Fischer, L. F., Soto-Albors, R. M., Buck, F. & Harnett, M. T. Representation of visual landmarks in retrosplenial cortex. eLife 9, e51458 (2020).
Miller, A. M. P., Mau, W. & Smith, D. M. Retrosplenial cortical representations of space and future goal locations develop with learning. Curr. Biol. 29, 2083–2090 (2019).
Jacob, P. Y. et al. An independent, landmark-dominated head-direction signal in dysgranular retrosplenial cortex. Nat. Neurosci. 20, 173–175 (2017).
Sit, K. K. & Goard, M. J. Coregistration of heading to visual cues in retrosplenial cortex. Nat. Commun. 14, 1992 (2023).
Fallahnezhad, M. et al. Cerebellar control of a unitary head direction sense. Proc. Natl. Acad. Sci. USA 120, e2214539120 (2023).
van Wijngaarden, J. B. G., Babl, S. S. & Ito, H. T. Entorhinal-retrosplenial circuits for allocentric-egocentric transformation of boundary coding. Elife 9, 1–25 (2020).
Alexander, A. S. et al. Egocentric boundary vector tuning of the retrosplenial cortex. Sci. Adv. 6, eaaz2322 (2020).
Cheng, N. et al. Egocentric processing of items in spines, dendrites, and somas in the retrosplenial cortex. Neuron 112, 646–660 (2024).
Vann, S. D. & Aggleton, J. P. Selective dysgranular retrosplenial cortex lesions in rats disrupt allocentric performance of the radial-arm maze task. Behavioral Neuroscience 119, 1682–1686 (2005).
Hindley, E. L., Nelson, A. J. D., Aggleton, J. P. & Vann, S. D. The rat retrosplenial cortex is required when visual cues are used flexibly to determine location. Behav. Brain Res. 263, 98–107 (2014).
Murakami, T., Yoshida, T., Matsui, T. & Ohki, K. Wide-field Ca2+ imaging reveals visually evoked activity in the retrosplenial area. Front. Mol. Neurosci. 8, 20 (2015).
Sugar, J., Witter, M. P., van Strien, N. M. & Cappaert, N. L. M. The retrosplenial cortex: Intrinsic connectivity and connections with the (para)hippocampal region in the rat. An interactive connectome. Front. Neuroinformatics 5, 7 (2011).
Mao, D., Kandler, S., McNaughton, B. L. & Bonin, V. Sparse orthogonal population representation of spatial context in the retrosplenial cortex. Nat. Commun. 8, 243 (2017).
Powell, A. et al. Stable encoding of visual cues in the mouse retrosplenial cortex. Cereb. Cortex 30, 4424–4437 (2020).
Van Groen, T. & Wyss, J. M. Connections of the retrosplenial granular b cortex in the rat. J. Comp. Neurol. 463, 249–263 (2003).
Li, Y. et al. Dissection of the long-range circuit of the mouse intermediate retrosplenial cortex. Commun. Biol. 8, 56 (2025).
Tsai, T. C., Yu, T. H., Hung, Y. C., Fong, L. I. & Sen, H. K. Distinct contribution of granular and agranular subdivisions of the retrosplenial cortex to remote contextual fear memory retrieval. J. Neurosci. 42, 877–893 (2022).
Yamawaki, N., Corcoran, K. A., Guedea, A. L., Shepherd, G. M. G. & Radulovic, J. Differential contributions of glutamatergic hippocampal→retrosplenial cortical projections to the formation and persistence of context memories. Cerebral Cortex 29, 2728–2736 (2019).
Trask, S. & Helmstetter, F. J. Unique roles for the anterior and posterior retrosplenial cortices in encoding and retrieval of memory for context. Cerebral Cortex 32, 3602–3610 (2022).
Franco, L. M. & Goard, M. J. A distributed circuit for associating environmental context with motor choice in retrosplenial cortex. Sci. Adv. 7, eabf9815 (2021).
Royer, S. et al. Control of timing, rate and bursts of hippocampal place cells by dendritic and somatic inhibition. Nat. Neurosci. 15, 769–775 (2012).
Lande, A. S., Garvert, A. C., Ebbesen, N. C., Jordbræk, S. V. & Vervaeke, K. Representations of tactile object location in the retrosplenial cortex. Curr. Biol. 33, 4599–4610 (2023).
Zhuang, J. et al. An extended retinotopic map of mouse cortex. eLife 6, e18372 (2017).
Andermann, M. L., Kerlin, A. M., Roumis, D. K., Glickfeld, L. L. & Reid, R. C. Functional specialization of mouse higher visual cortical areas. Neuron 72, 1025–1039 (2011).
Han, X., Vermaercke, B. & Bonin, V. Diversity of spatiotemporal coding reveals specialized visual processing streams in the mouse cortex. Nat. Commun. 13, 3249 (2022).
de Vries, S. E. J. et al. A large-scale standardized physiological survey reveals functional organization of the mouse visual cortex. Nature Neurosci. 23, 138–151 (2020).
Marshel, J. H., Garrett, M. E., Nauhaus, I. & Callaway, E. M. Functional specialization of seven mouse visual cortical areas. Neuron 72, 1040–1054 (2011).
Yamawaki, N. et al. Long-range inhibitory intersection of a retrosplenial thalamocortical circuit by apical tuft-targeting CA1 neurons. Nat. Neurosci. 22, 618–626 (2019).
Wang, Q. & Burkhalter, A. Area map of mouse visual cortex. J. Compar. Neurol. 502, 339–357 (2007).
Wang, Q. et al. The allen mouse brain common coordinate framework: a 3D reference atlas. Cell 181, 936–953 (2020).
Li, Z. et al. Direction-selective laterodorsal thalamic nucleus encodes optic flow and turning in spatial navigation. Neuron 113, 3424–3440 (2025).
Gianatti, M. et al. Multiple long-range projections convey position information to the agranular retrosplenial cortex. Cell Rep. 42, 113109 (2023).
Mao, D. et al. Hippocampus-dependent emergence of spatial sequence coding in retrosplenial cortex. Proc. Natl. Acad. Sci. USA 115, 8015–8018 (2018).
Lever, C., Burton, S., Jeewajee, A., O’Keefe, J. & Burgess, N. Boundary vector cells in the subiculum of the hippocampal formation. J. Neurosci. 29, 9771–9777 (2009).
Sun, Y., Nitz, D. A., Xu, X. & Giocomo, L. M. Subicular neurons encode concave and convex geometries. Nature 627, 821–829 (2024).
Nitzan, N. et al. Propagation of hippocampal ripples to the neocortex by way of a subiculum-retrosplenial pathway. Nat. Commun. 11, 1947 (2020).
Fanselow, M. S. & Dong, H. W. Are the dorsal and ventral hippocampus functionally distinct structures? Neuron 65, 7–19 (2010).
Sullivan, K. E. et al. Sharp cell-type-identity changes differentiate the retrosplenial cortex from the neocortex. Cell Rep. 42, 112206 (2023).
LaChance, P. A. & Taube, J. S. The anterior thalamus preferentially drives allocentric but not egocentric orientation tuning in postrhinal cortex. J. Neurosci. 44, e0861232024 (2024).
Roy, D. S. et al. Anterior thalamic circuits crucial for working memory. Proc. Natl. Acad. Sci. USA 119, e2118712119 (2022).
Jankowski, M. M. et al. The anterior thalamus provides a subcortical circuit supporting memory and spatial navigation. Front. Syst. Neurosci. 7, 45 (2013).
Dana, H. et al. Thy1-GCaMP6 transgenic mice for neuronal population imaging in vivo. PloS ONE 9, e108697 (2014).
Wekselblatt, J. B., Flister, E. D., Piscopo, D. M. & Niell, C. M. Large-scale imaging of cortical dynamics during sensory perception and behavior. J. Neurophysiol. 115, 2852–2866 (2016).
Goldey, G. J. et al. Removable cranial windows for long-term imaging in awake mice. Nat. Protoc. 9, 2515–2538 (2014).
Peirce, J. W. PsychoPy-psychophysics software in Python. J. Neurosci. Methods 162, 8–13 (2007).
Conte, W. L., Kamishina, H. & Reep, R. L. Multiple neuroanatomical tract-tracing using fluorescent Alexa Fluor conjugates of cholera toxin subunit B in rats. Nat. Protoc. 4, 1157–1166 (2009).
Stringer, C. et al. Extracting large-scale neural activity with Suite2p. Preprint at https://doi.org/10.64898/2026.02.04.703741 (2026).
Friedrich, J., Zhou, P. & Paninski, L. Fast online deconvolution of calcium imaging data. PLoS Comput. Biol. 13, e1005423 (2017).
Kropff, E., Carmichael, J. E., Moser, M. B. & Moser, E. I. Speed cells in the medial entorhinal cortex. Nature 523, 419–424 (2015).
Davidson, T. J., Kloosterman, F. & Wilson, M. A. Hippocampal replay of extended experience. Neuron 63, 497–507 (2009).
Zhang, K., Ginzburg, I., McNaughton, B. L. & Sejnowski, T. J. Interpreting neuronal population activity by reconstruction: unified framework with application to hippocampal place cells. Physical Variables. J. Neurophysiol. 79, 1017–1044 (1998).
Danielson, N. B. et al. Sublayer-specific coding dynamics during spatial navigation and learning in hippocampal area CA1. Neuron 91, 652–665 (2016).
Mao, D., Molina, L. A., Bonin, V. & McNaughton, B. L. Vision and locomotion combine to drive path integration sequences in mouse retrosplenial cortex. Curr. Biol. 30, 1680–1688 (2020).
Skaggs, W. E., McNaughton, B. L., Gothard, K. M. & Markus, E. J. An information-theoretic approach to deciphering the hippocampal code. Adv. Neural Inf. Process. Syst. 5, 1030–1037 (1993).
Ohki, K., Chung, S., Ch, Y. H., Kara, P. & Clay Reid, R. Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex. Nature 433, 697–603 (2005).
Shamash, P., Carandini, M., Harris, K. D. & Steinmetz, N. A. A tool for analyzing electrode tracks from slice histology. Preprint at https://doi.org/10.1101/447995 (2018).
Claudi, F. et al. Visualizing anatomically registered data with brainrender. eLife 10, e65751 (2021).
Acknowledgements
We thank Ben Vermaercke, Adrien Philippon, and Ta-Shun Su for technical assistance. YT.W. was supported by a scholarship from the Taiwanese Ministry of Education (MOE)–KU Leuven. J.C. acknowledges support from FWO (Fellowship 1226320N). V.B. acknowledges support from FWO (grants G0C1220N, G083Y24N, and G056725N). V.B. and F.K. acknowledge support from FWO (grant G077321N). We are also grateful to the members of the Bonin and Kloosterman labs for valuable discussions and feedback.
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Conceptualization: YT.W., J.C., and V.B.; Methodology: YT.W. and V.B.; Software: YT.W.; Investigation: YT.W., V.B., and F.K.; Data Analysis: YT.W.; Visualization: YT.W. Writing—original draft: YT.W.; Writing—review & editing: YT.W., V.B., and F.K.; Funding acquisition: J.C., V.B., and F.K.; Supervision: V.B. and F.K.
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Wei, YT., Couto, J., Kloosterman, F. et al. Anterior and posterior retrosplenial cortex form distinct visuospatial circuits in the mouse. Nat Commun (2026). https://doi.org/10.1038/s41467-026-70762-z
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DOI: https://doi.org/10.1038/s41467-026-70762-z
