Previous Ketamine Produces an Enduring Blockade of Neurochemical and Behavioral Effects of Uncontrollable Stress.

Recent interest in the antidepressant and anti-stress effects of subanesthetic doses of ketamine, an NMDA receptor antagonist, has identified mechanisms whereby ketamine reverses the effect of stress, but little is known regarding the prophylactic effect ketamine might have on future stressors. Here we investigate the prophylactic effect of ketamine against neurochemical and behavioral changes that follow inescapable, uncontrollable tail shocks (ISs) in Sprague Dawley rats. IS induces increased anxiety, which is dependent on activation of serotonergic (5-HT) dorsal raphe nucleus (DRN) neurons that project to the basolateral amygdala (BLA). Ketamine (10 mg/kg, i.p.) administered 2 h, 1 week, or 2 weeks before IS prevented the increased extracellular levels of 5-HT in the BLA typically produced by IS. In addition, ketamine administered at these time points blocked the decreased juvenile social investigation produced by IS. Microinjection of ketamine into the prelimbic (PL) region of the medial prefrontal cortex duplicated the effects of systemic ketamine, and, conversely, systemic ketamine effects were prevented by pharmacological inhibition of the PL. Although IS does not activate DRN-projecting neurons from the PL, IS did so after ketamine, suggesting that the prophylactic effect of ketamine is a result of altered functioning of this projection. SIGNIFICANCE STATEMENT: The reported data show that systemic ketamine, given up to 2 weeks before a stressor, blunts behavioral and neurochemical effects of the stressor. The study also advances understanding of the mechanisms involved and suggests that ketamine acts at the prelimbic cortex to sensitize neurons that project to and inhibit the DRN.

The role of prior stressor controllability and the dorsal raphé nucleus in sucrose preference and social exploration.

Research investigating how control over stressors affects behavior often utilizes freezing and shuttle escape learning as the behavioral endpoints. These endpoints have been argued to reflect anxious or depressed states, but these descriptions are problematic. The present study sought to determine the impact of stressor controllability and the dorsal raphé nucleus (DRN) on sucrose preference and juvenile social exploration, putative measures of anhedonia and anxiety that are commonly used in studies of stress per se. In Experiment 1 rats were exposed to escapable stress (ES) or yoked-inescapable stress (IS) tailshocks. In Experiment 2 ES or IS was given 7 days before all rats received IS. In Experiment 3 the DRN was inactivated during IS by microinjection of 8-OH-DPAT. Sucrose preference and social exploration were tested for several days after stress. A fourth experiment confirmed that juvenile social exploration is sensitive to traditional beta-carboline and benzodiazepine manipulations. Both ES and IS reduced sucrose preference, but only IS reduced social exploration. Prior treatment with ES prevented the effect of IS on social exploration but did not prevent the effect of IS on sucrose preference and inactivation of the DRN prevented the effect of IS on social exploration but did not change sucrose preference. The present results indicate that social exploration but not sucrose preference is sensitive to prior stressor controllability, and that DRN activation mediates the effect of IS on social exploration. We argue that DRN-5-HT activation mediates a state of generalized anxiety produced by uncontrollable stress and that juvenile social exploration is a useful behavioral endpoint in stressor controllability studies.

The effects of an anabolic androgenic steroid and low serotonin on social and non-social behaviors in male rats.

The behavioral and neurochemical impact of low serotonin (5-HT) was examined in gonadally intact male rats exposed to an anabolic androgenic steroid (AAS) during puberty. Low 5-HT was induced beginning on postnatal day 26 using parachlorophylalanine (PCPA). Injections of the AAS, testosterone (TP), began on day 40. The rats were tested in both non-social (locomotor activity and nose poke for food) and social (low-threat and high-threat) contexts. PCPA and TP+PCPA significantly decreased locomotor activity. PCPA alone significantly increased nose poke latency compared to controls. Freezing in the PCPA group was significantly elevated compared to TP and TP+PCPA groups, but not compared to controls. AAS did not affect non-social behaviors. Thus, low serotonin may increase freezing in a non-social context. Following provocation, PCPA and TP+PCPA significantly increased aggression toward smaller non-threatening opponents, suggesting that males with low 5-HT are more aggressive in a low-threat context when provoked. In the resident-pair intruder test, TP significantly increased aggression whereas PCPA did not, suggesting that in a high-threat context, aggression is primarily mediated by AAS. TP+PCPA males were also significantly more aggressive in the high-threat context suggesting that exposure to AAS may override freezing behavior induced by low serotonin. Both PCPA and TP+PCPA significantly and substantially depleted 5-HT and 5-HIAA in all brain regions examined. AAS significantly decreased 5-HIAA levels in the hypothalamus and increased 5-HT levels in the frontal cortex. Following withdrawal from TP+PCPA, most behavioral and neurochemical measures returned to control levels. These data suggest that low serotonin may be a contributing factor in the increased aggression displayed by adolescents who abuse AAS.

Two models of inescapable stress increase tph2 mRNA expression in the anxiety-related dorsomedial part of the dorsal raphe nucleus.

Expression of TPH2, the rate-limiting enzyme for brain serotonin synthesis, is elevated in the dorsal raphe nucleus (DR) of depressed suicide victims. One hypothesis is that this increase in TPH2 expression is stress-induced. Here, we used an established animal model to address whether exposure to an acute stressor, inescapable tail shock (IS), increases tph2 mRNA and Tph2 protein expression, and if IS sensitizes the DR to a subsequent, heterotypic stressor. In Experiment 1, we measured tph2 mRNA expression 4 h after IS or home cage (HC) control conditions in male rats, using in situ hybridization histochemistry. In Experiment 2, we measured Tph2 protein expression 12 h or 24 h after IS using western blot. In Experiment 3, we measured tph2 mRNA expression following IS on Day 1, and cold swim stress (10 min, 15 °C) on Day 2. Inescapable tail shock was sufficient to increase tph2 mRNA expression 4 h and 28 h later, selectively in the dorsomedial DR (caudal aspect of the dorsal DR, cDRD; an area just rostral to the caudal DR, DRC) and increased Tph2 protein expression in the DRD (rostral and caudal aspects of the dorsal DR combined) 24 h later. Cold swim increased tph2 mRNA expression in the dorsomedial DR (cDRD) 4 h later. These effects were associated with increased immobility during cold swim, elevated plasma corticosterone, and a proinflammatory plasma cytokine milieu (increased interleukin (IL)-6, decreased IL-10). Our data demonstrate that two models of inescapable stress, IS and cold swim, increase tph2 mRNA expression selectively in the anxiety-related dorsomedial DR (cDRD).

Short- and long-term consequences of stressor controllability in adolescent rats.

Adolescence is a developmental period in which brain structures involved with stress responses, such as the medial pre-frontal cortex (mPFC), mature. Therefore, exposure to a stressor at this time may have effects that endure the lifespan. The goal of the present study was to determine whether behavioral control over an adolescent stressor mitigates the behavioral and neurochemical consequences of the stressor as occurs in adult rats. Adolescent rats (post natal day 35) were exposed to either inescapable (IS) or escapable tailshocks (ES). As in adults we observed a "stressor controllability effect"; IS reduced social exploration and activated the serotonergic dorsal raphé nucleus while ES did not. Excitotoxic lesions of the medial prefrontal cortex prevented the stressor controllability effect. We also demonstrate that a controllable adolescent stress prevents the behavioral and neurochemical consequences of IS in adulthood. Thus, the controllability of a stressor during adolescence is an important psychological factor.

Active behavioral coping alters the behavioral but not the endocrine response to stress.

Exposure to traumatic stressors typically causes lasting changes in emotionality and behavior. However, coping strategies have been shown to prevent and alleviate many stress consequences and the biological mechanisms that underlie coping are of great interest. Whereas the laboratory stressor inescapable tail-shock induces anxiety-like behaviors, here we demonstrate that permitting a rat to chew on a wooden dowel during administration of tail-shock prevented the development of anxiety like behaviors in the open field and juvenile social exploration tests. Uncontrollable stressors increase corticosterone and decrease thyroid hormone, and we hypothesized that coping would blunt these changes. While tail-shock did produce these effects, active coping did not alter hormone levels. The dissociation between behavioral resilience and circulating hormones is discussed with regard to the utility of these molecules as biomarkers for psychiatric disease.