Layer-specific input to medial prefrontal cortex is linked to stress susceptibility.

Stress response is essential for adapting to an ever-changing environment. However, the mechanisms that render some individuals susceptible to stress are poorly understood. While chronic stress is known to induce dendritic atrophy and spine loss in medial prefrontal cortex (mPFC), its impact on synapses made by long-range projections terminating on the mPFC remains unknown. Here, we labeled synapses on male mouse mPFC dendrites formed by ventral hippocampus (VH), basolateral amygdala (BLA) and ventral tegmental area (VTA) long-range afferents using different-colored eGRASP constructs. We obtained multispectral 3D-images of the mPFC covering all cortical laminae, and automatically segmented the dendrites and synapses. In layer II/III, the relative abundances and spatial organizations of VH-mPFC and BLA-mPFC synapses changed similarly in stress resilient (SR) and stress susceptible (SS) mice when compared to stress naïve (SN) mice. In layers Vb and VI, on the other hand, the percentage of BLA-mPFC synapses increased and that of VH-mPFC decreased only in SS mice. Moreover, the distances of VH synapses to their corresponding closest BLA synapses decreased and the distances of BLA synapses to their corresponding closest VH synapses increased in the SS group. Consistently, the percentage of single dendritic segments receiving input from multiple brain regions increased in the SS group, suggesting that long-range synaptic inputs to deep layers of mPFC were disorganized in SS mice. Our findings demonstrate afferent- and lamina-specific differential reorganization of synapses between different stress phenotypes, suggesting specific roles for different long-range projections in mediating the stress response.