4.7 Article

Single-cell reconstruction reveals input patterns and pathways into corticotropin-releasing factor neurons in the central amygdala in mice

Journal

COMMUNICATIONS BIOLOGY
Volume 5, Issue 1, Pages -

Publisher

NATURE PORTFOLIO
DOI: 10.1038/s42003-022-03260-9

Keywords

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Funding

  1. National Natural Science Foundation of China [32030046]
  2. Strategic Priority Research Program of the Chinese Academy of Science [XDB32020200]
  3. National Key R&D Program of China [2016YFC1305900]

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This study used a rabies-virus system to map the single-cell input onto corticotropin-releasing factor (CRF) neurons in the central amygdala (CeA). The inputs were found to be located in 44 nested subregions, with most of them being top-down convergent inputs from the cortex, especially in layer 4 of the somatosensory cortex, and relaying information from the thalamus. The bottom-up divergent inputs had a higher expression of glutamate decarboxylase. The study also revealed the en passant structures of single input neurons through in-situ reconstruction. These findings provide valuable insights into the circuit mechanisms of CeA-CRF neurons.
Corticotropin-releasing factor (CRF) neurons are one of the most densely distributed cell types in the central amygdala (CeA), and are involved in a wide range of behaviors including anxiety and learning. However, the fundamental input circuits and patterns of CeA-CRF neurons are still unclear. Here, we generate a monosynaptic-input map onto CeA-CRF neurons at single-cell resolution via a retrograde rabies-virus system. We find all inputs are located in 44 nested subregions that directly innervate CeA-CRF neurons; most of them are top-down convergent inputs expressing Ca2+/calmodulin-dependent protein kinase II, and are centralized in cortex, especially in the layer 4 of the somatosensory cortex, which may directly relay information from the thalamus. While the bottom-up divergent inputs have the highest proportion of glutamate decarboxylase expression. Finally, en passant structures of single input neuron are revealed by in-situ reconstruction in a modified 3D-reference atlas, represented by a Periaqueductal gray-Subparafascicular nucleus-Subthalamic nucleus-Globus pallidus-Caudoputamen-CeA pathway. Taken together, our findings provide morphological and connectivity properties of inputs onto CeA-CRF neurons, which may provide insights for future studies interrogating circuit mechanisms of CeA-CRF neurons in mediating various functions. Viral retrograde tracing identifies input regions and patterns into the corticotropin releasing factor-expressing neurons in central amygdala, providing an important resource to disentangle the role of these cells in fear and anxiety.

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