Inhibitory regulation of calcium transients in prefrontal dendritic spines is compromised by a nonsense Shank3 mutation.

The SHANK3 gene encodes a postsynaptic scaffold protein in excitatory synapses, and its disruption is implicated in neurodevelopmental disorders such as Phelan-McDermid syndrome, autism spectrum disorder, and schizophrenia. Most studies of SHANK3 in the neocortex and hippocampus have focused on disturbances in pyramidal neurons. However, GABAergic interneurons likewise receive excitatory inputs and presumably would also be a target of constitutive SHANK3 perturbations. In this study, we characterize the prefrontal cortical microcircuit in awake mice using subcellular-resolution two-photon microscopy. We focused on a nonsense R1117X mutation, which leads to truncated SHANK3 and has been linked previously to cortical dysfunction. We find that R1117X mutants have abnormally elevated calcium transients in apical dendritic spines. The synaptic calcium dysregulation is due to a loss of dendritic inhibition via decreased NMDAR currents and reduced firing of dendrite-targeting somatostatin-expressing (SST) GABAergic interneurons. Notably, upregulation of the NMDAR subunit GluN2B in SST interneurons corrects the excessive synaptic calcium signals and ameliorates learning deficits in R1117X mutants. These findings reveal dendrite-targeting interneurons, and more broadly the inhibitory control of dendritic spines, as a key microcircuit mechanism compromised by the SHANK3 dysfunction.

Sex differences in cognitive flexibility are driven by the estrous cycle and stress-dependent.

Stress is associated with psychiatric disorders such as post-traumatic stress disorder, major depressive disorder, anxiety disorders, and panic disorders. Women are more likely to be diagnosed with these stress-related psychiatric disorders than men. A key phenotype in stress-related psychiatric disorders is impairment in cognitive flexibility, which is the ability to develop new strategies to respond to different patterns in the environment. Because gonadal hormones can contribute to sex differences in response to stress, it is important to consider where females are in their cycle when exposed to stress and cognitive flexibility testing. Moreover, identifying neural correlates involved in cognitive flexibility could not only build our understanding of the biological mechanisms behind this crucial skill but also leads to more targeted treatments for psychiatric disorders. Although previous studies have separately examined sex differences in cognitive flexibility, stress effects on cognitive flexibility, and the effect of gonadal hormones on cognitive flexibility, many of the findings were inconsistent, and the role of the estrous cycle in stress-induced impacts on cognitive flexibility is still unknown. This study explored potential sex differences in cognitive flexibility using an operant strategy shifting-paradigm after either control conditions or restraint stress in freely cycling female and male rats (with estrous cycle tracking in the female rats). In addition, we examined potential neural correlates for any sex differences observed. In short, we found that stress impaired certain aspects of cognitive flexibility and that there were sex differences in cognitive flexibility that were driven by the estrous cycle. Specifically, stress increased latency to first press and trials to criterion in particular tasks. The female rats demonstrated more omissions and perseverative errors than the male rats; the sex differences were mostly driven by proestrus female rats. Interestingly, the number of orexinergic neurons was higher in proestrus female rats than in the male rats under control conditions. Moreover, orexin neural count was positively correlated with number of perseverative errors made in cognitive flexibility testing. In sum, there are sex differences in cognitive flexibility that are driven by the estrous cycle and are stress-dependent, and orexin neurons may underlie some of the sex differences observed.

Ketamine disinhibits dendrites and enhances calcium signals in prefrontal dendritic spines.

A subanesthetic dose of ketamine causes acute psychotomimetic symptoms and sustained antidepressant effects. In prefrontal cortex, the prevailing disinhibition hypothesis posits that N-methyl-d-aspartate receptor (NMDAR) antagonists such as ketamine act preferentially on GABAergic neurons. However, cortical interneurons are heterogeneous. In particular, somatostatin-expressing (SST) interneurons selectively inhibit dendrites and regulate synaptic inputs, yet their response to systemic NMDAR antagonism is unknown. Here, we report that ketamine acutely suppresses the activity of SST interneurons in the medial prefrontal cortex of the awake mouse. The deficient dendritic inhibition leads to greater synaptically evoked calcium transients in the apical dendritic spines of pyramidal neurons. By manipulating NMDAR signaling via GluN2B knockdown, we show that ketamine's actions on the dendritic inhibitory mechanism has ramifications for frontal cortex-dependent behaviors and cortico-cortical connectivity. Collectively, these results demonstrate dendritic disinhibition and elevated calcium levels in dendritic spines as important local-circuit alterations driven by the administration of subanesthetic ketamine.