Ketamine ameliorates activity-based anorexia of adolescent female mice through changes in the prevalence of NR2B-containing NMDA receptors at excitatory synapses that are in opposite directions for of pyramidal neurons versus GABA interneurons in medial prefrontal cortex

(250 word limit) A previous study showed that a single sub-anesthetic dose of ketamine (30 mg/kg-KET, IP) has an immediate and long-lasting (>20 days) effect of reducing maladaptive behaviors associated with activity-based anorexia (ABA) among adolescent female mice. This study sought to determine whether synaptic plasticity involving NR2B-containing NMDA receptors (NR2B) at excitatory synapses in the prelimbic region of medial prefrontal cortex (mPFC) contributes to this ameliorative effect. To this end, quantitative electron microscopic analyses of NR2B-subunit immunoreactivity at excitatory synapses of pyramidal neurons (PN) and GABAergic interneurons (GABA-IN) were conducted upon layer 1 of mPFC of the above-described mice that received a single efficacious 30 mg/kg-KET (N=8) versus an inefficacious 3 mg/kg-KET (N=8) dose during the food-restricted day of the first ABA induction (ABA1). Brain tissue was collected after these animals underwent recovery from ABA1, then of recovery from a second ABA induction (ABA2), 22 days after the ketamine injection. For all three parameters used to quantify ABA resilience (increased food consumption, reduced wheel running, body weight gain), 30 mg/kg-KET evoked synaptic plasticity in opposite directions for PN and GABA-IN, with changes at excitatory synapses on GABA-IN dominating the adaptive behaviors more than on PN. The synaptic changes were in directions consistent with changes in the excitatory outflow from mPFC that weaken food consumption-suppression, strengthen wheel running suppression and enhance food consumption. We hypothesize that 30 mg/kg-KET promotes these long-lasting changes in the excitatory outflow from mPFC after acutely blocking the hunger and wheel-access activated synaptic circuits underlying maladaptive behaviors during ABA.