The role of different amygdala nuclei (neuroanatomical subdivisions) in processing Pavlovian conditioned fear has been studied extensively, but the function of the heterogeneous neuronal subtypes within these nuclei remains poorly understood. Here we use molecular genetic approaches to map the functional connectivity of a subpopulation of GABA-containing neurons, located in the lateral subdivision of the central amygdala (CEl), which express protein kinase C-δ (PKC-δ). Channelrhodopsin-2-assisted circuit mapping in amygdala slices and cell-specific viral tracing indicate that PKC-δ+ neurons inhibit output neurons in the medial central amygdala (CEm), and also make reciprocal inhibitory synapses with PKC-δ− neurons in CEl. Electrical silencing of PKC-δ+ neurons in vivo suggests that they correspond to physiologically identified units that are inhibited by the conditioned stimulus, called CEloff units. This correspondence, together with behavioural data, defines an inhibitory microcircuit in CEl that gates CEm output to control the level of conditioned freezing.
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We thank N. Heinz and X. Gong for generating BAC transgenic mice; C. Saper for providing the Cre-dependent hrGFP AAV construct; C. Xiao for training in slice electrophysiology and preliminary experiments; H. Lester for advice on the GluCl system; L. van Tright for performing in situ hybridizations; W. Lerchner for providing a CAG-driven GluClβ construct; A. Chang for help with behavioural scoring; M. Martinez for tail genotyping; G. Mosconi for laboratory management; and J. Alex, R. Bayon and R. Sauza for animal care. This work was supported by NIH grant 1 R01 MH085082-01A1 and by funds from the Caltech ‘Conscious Mouse’ project. W.H. was supported by a fellowship of the Human Frontier Science Program and P.S.K. by the Jane Coffin Childs Memorial Fund for Medical Research. S.C. and A.L. were supported by the Novartis Research Foundation. D.J.A. is an Investigator of the Howard Hughes Medical Institute.
The authors declare no competing financial interests.
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Haubensak, W., Kunwar, P., Cai, H. et al. Genetic dissection of an amygdala microcircuit that gates conditioned fear. Nature 468, 270–276 (2010). https://doi.org/10.1038/nature09553
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