Article

A distinct entorhinal cortex to hippocampal CA1 direct circuit for olfactory associative learning

  • Nature Neuroscience volume 20, pages 559570 (2017)
  • doi:10.1038/nn.4517
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Abstract

Lateral and medial parts of entorhinal cortex (EC) convey nonspatial 'what' and spatial 'where' information, respectively, into hippocampal CA1, via both the indirect EC layer 2→ hippocampal dentate gyrus→CA3→CA1 and the direct EC layer 3→CA1 paths. However, it remains elusive how the direct path transfers distinct information and contributes to hippocampal learning functions. Here we report that lateral EC projection neurons selectively form direct excitatory synapses onto a subpopulation of morphologically complex, calbindin-expressing pyramidal cells (PCs) in the dorsal CA1 (dCA1), while medial EC neurons uniformly innervate all dCA1 PCs. Optogenetically inactivating the distinct lateral EC–dCA1 connections or the postsynaptic dCA1 calbindin-expressing PC activity slows olfactory associative learning. Moreover, optetrode recordings reveal that dCA1 calbindin-expressing PCs develop more selective spiking responses to odor cues during learning. Thus, our results identify a direct lateral EC→dCA1 circuit that is required for olfactory associative learning.

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Acknowledgements

We thank R. Hou (ION) for her help with the behavioral test; B. Wang and Y.Shu (BNU) for their help with single-cell RT-PCR; J.Z. Huang (CSHL) for providing the Calb2-IRES-Cre mice; Z. Qiu (ION) for providing AAV-hSyn-ChR2-mCherry vectors; and J.J. Knierim (JHU), L. Zhang and H. Tao (USC), Y.X. Lin (MIT) and B. Li (CSHL) for their critical comments on the manuscript. This work was supported by grants from the State Key Research Program of China (2011CBA00404 to X.Z.), the Basic Research Project of Shanghai Science and Technology Commission (No. 15JC1400102 to L.L.) and the Natural Science Foundation of China (NSFC 81327802 to S.Z. and Y. Liu).

Author information

Author notes

    • Yiding Li
    •  & Jiamin Xu

    These authors contributed equally to this work.

Affiliations

  1. Institute of Neuroscience, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.

    • Yiding Li
    • , Jia Zhu
    •  & Chengyu Li
  2. State Key Laboratory of Cognitive Neuroscience & Learning and IDG/McGovern Institute for Brain Research, Beijing Normal University, Beijing, China.

    • Yiding Li
    • , Nan Liu
    • , Malte J Rasch
    • , Xiang Gu
    • , Xiang Li
    •  & Xiaohui Zhang
  3. University of Chinese Academy of Sciences, Shanghai, China.

    • Yiding Li
    •  & Jia Zhu
  4. Key Laboratory of Brain Functional Genomics-Ministry of Education, School of Life Science, East China Normal University, Shanghai, China.

    • Jiamin Xu
    •  & Longnian Lin
  5. Britton Chance Center for Biomedical Photonics and Department of Biomedical Engineering, Wuhan National Laboratory for Optoelectronics–Huazhong University of Science and Technology, Wuhan, China.

    • Yafeng Liu
    •  & Shaoqun Zeng
  6. State Key Laboratory of Virology, CAS Center for Excellence in Brain Science and Intelligence Technology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.

    • Wenbo Zeng
    • , Haifei Jiang
    •  & Minhua Luo
  7. Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education and State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing, China.

    • Ning Huang
    • , Junlin Teng
    •  & Jianguo Chen

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Contributions

X.Z. conceived and supervised the project, and X.Z. and Y. Li designed the experiments. Y. Li performed the virus injection, in vitro electrophysiology, immunostaining, single-cell RT-PCR, cellular imaging, morphological reconstruction and behavioral tests. J.X. performed the in vivo optetrode recording, spike sorting and place field analysis. N.L. and M.J.R. performed the TREE analysis, cluster analysis and computational simulation. W.Z., H.J. and M.L. designed and prepared the HSV virus. N.H., J.T. and J.C. designed and conducted the EM experiment. Y. Liu and S.Z. designed and built the AOD-based rapid laser stimulation system. J.Z. and C.L. participated in the behavioral experiment. X.G. and X.L. prepared and genotyped all transgenic mice. Y. Li, L.L. and X.Z. analyzed the data. X.Z. and Y. Li wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Xiaohui Zhang.

Integrated supplementary information

Supplementary figures

  1. 1.

    Analyses of dCA1 PC clusters by the Jump method, and principle component analysis (PCA) on different morphological parameters.

  2. 2.

    Characterization of intrinsic action potential firing patterns and excitability of dCA1 sPCs and cPCs.

  3. 3.

    Differential propagation of dendritic excitation to the soma in sPCs and cPCs revealed by TREES and NEURON simulations.

  4. 4.

    Histological characterization of viral expression of ChR2 in the LEC, MEC and their axon projections along the anterior–posterior axis of CA1.

  5. 5.

    Characterization of optogenetic activation of EC–dCA1 monosynaptic transmission by AOD-based laser stimulation system.

  6. 6.

    Direct long-range GABAergic transmission from LEC and MEC to dCA1 pyramidal neurons.

  7. 7.

    Comparison of transmission strength of CA3 SC excitatory synapses on dCA1 sPCs and cPCs.

  8. 8.

    Characterization of Calb2-IRES-Cre::Ai9 and Calb1-2A-dgCre::Ai9 mice.

  9. 9.

    HSV-1-G3 anterograde transportation from dorsal or ventral LEC and MEC to hippocampal CA1.

  10. 10.

    Postbehavior histological characterization of NpHR-EYFP expression in LEC or MEC and the locations of implanted optical fibers or optetrodes in dCA1.

  11. 11.

    Similar impairments of reversed learning by inactivation of LEC–dCA1 transmissions or of postsynaptic dCA1 Calb+ cPCs.

  12. 12.

    Optogenetic tagging of dCA1 Calb+ cPC units with Arch in vivo in comparison with that of tagging with ChR2.

  13. 13.

    Varied spiking responses and selectivities in response to odors A and B during odor sampling in trials.

  14. 14.

    Comparison of neuronal spiking activities in Correct and Error trials in the behavioral task.

  15. 15.

    Comparison of spontaneous firing rates during different brain states between the identified Calb+ and Calb dCA1 PCs.

Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–15 and Supplementary Tables 1–16

  2. 2.

    Supplementary Methods Checklist