Howard S. Stock

Differential release of corticotropin-releasing hormone (CRH) in the amygdala during different types of stressors

The purpose of this study was to determine if differences exist between the effects of acute treadmill running and restraint stress on corticotropin-releasing hormone (CRH) release within the amygdala of rats. Extracellular CRH immunoreactivity (CRH-IR) was measured in microdialysate collected from the central nucleus of the amygdala (CeA) during exposure to an inactivated treadmill (TC), during 1 h treadmill running to exhaustion (RUN), and 1 h restraint (RES). Extracellular CRH-IR increased from control levels during the first 20-min period for TC, RUN, and RES, with the largest increase during RES. During the second 20-min period, only RES maintained levels higher than control values. CRH release was higher than control during the third 20-min period of RES and RUN. A second experiment consisted of four groups of either cage controls (CC), TC, RUN, or RES. Immediately following the 60-min treatment, brains were removed and trunk blood collected for analysis of tissue CRH-IR and plasma corticosterone. While amygdala tissue CRH-IR was not different in the CC, TC and RUN rats, these groups had significantly lower levels than the RES animals. Hypothalamic tissue CRH-IR was not different between the CC and TC rats, but the levels were significantly higher in the RES and RUN rats than in the two control groups. Plasma corticosterone levels were elevated only in RES and RUN rats. Results from tissue analysis indicate that increased tissue CRH-IR in the amygdala and hypothalamus can be elicited by RES, while only the hypothalamus shows an increase following RUN. Further, extracellular CRH release in the CeA is increased throughout the period of RES, when rats are placed on the treadmill, and when the animals are approaching physical exhaustion. No increase is observed during the running period between placement on the treadmill and intense exertion. Overall, the data suggest that amygdala CRH release is regulated differently during treadmill running and restraint.

A lack of tolerance to the anxiolytic effects of diazepam on the plus-maze: comparison of male and female rats

Abstract Rationale: The demonstration of tolerance to the anxiolytic effects of benzodiazepines remains inconsistent. Objectives: The present study tested the hypothesis that intact and gonadectomized male and female rats might exhibit differential tolerance to the anxiolytic effects of diazepam (DZ). Methods: Following acute (3 days) or chronic (3 weeks) DZ exposure, all animals were tested on the elevated plus-maze and immediately sacrificed for analysis of corticosterone, adrenocorticotropin hormone, estrogen and progesterone levels in serum. In experiment 2, following acute or chronic DZ exposure, animals were treated with a DZ challenge dose on the test day. Results: In experiment 1, both acute and chronic DZ treatment similarly enhanced percentage open arm time and entries, regardless of the hormonal status of the animal. The results of experiment 2 showed that both acute and chronic DZ-treated animals exhibited a significantly higher percentage open arm time than control animals after the DZ challenge dose, and males and females did not differ in their responses to DZ exposure. Conclusions: The findings from these experiments suggest that tolerance to the anxiolytic effects of DZ did not develop in males or females, and that the hormonal status of the animal does not significantly alter the anxiolytic effects of DZ following either acute or chronic exposure. Following plus-maze exposure, females had significantly higher corticosterone levels than males and acute DZ treatment diminished this stress response.

Gender and gonadal hormone effects in the olfactory bulbectomy animal model of depression

Major depressive disorder (MDD) affects women to a greater extent then men; however, the few studies that have examined the role of gender in an animal model of depression have shown inconsistent results. The purpose of the present study was to determine if the gonadal hormone milieu of the animal modulated behavioral changes following olfactory bulbectomy (OBX), a well-documented animal model of depression. Body weight, sucrose preference levels and open-field activity levels were measured once a week for a period of 2 weeks in gonadally intact and gonadectomized male and female rats. Following these baseline measurements, animals underwent either OBX or sham surgery. Body weight, sucrose preference and activity levels were assessed for 4 weeks post-OBX surgery. OBX-gonadectomized animals exhibited higher activity levels than OBX gonadally intact and control animals. This effect of gonadectomy was more robust in males. OBX-females (both intact and gonadectomized) exhibited significantly lower sucrose preference levels than OBX-males (both intact and gonadectomized) and control animals. These results suggest that the gonadal hormone milieu of the animal plays a role in modulating sucrose preference and activity levels following OBX.

Changes in defensive behaviors following olfactory bulbectomy in male and female rats

The present study examined if olfactory bulbectomy (OBX) altered defensive behaviors on the elevated plus-maze and the open-field differently in male and female rats. Similar increases in defensive behaviors in male and female rats were observed in both tests following OBX. No significant correlations were detected between defensive behaviors and activity, supporting the hypothesis that some behavioral changes following OBX may be due to decreased defensive behaviors and not increased activity.

The conditioning of dyspneic suffocation fear. Effects of carbon dioxide concentration on behavioral freezing and analgesia.

Previous studies in our laboratory have shown that a single exposure to 100% carbon dioxide (CO2) can serve as an effective unconditioned stimulus (US) in a Pavlovian aversive-context conditioning paradigm in rats. Although the US exposure parameters employed in the initial studies were sufficient for producing a context-specific enhancement of behavioral freezing and analgesia, it had yet to be determined whether variations of these CO2 conditioning procedures would produce other conditioning effects. Thus, the purpose of the following experiment was to investigate the intensity of the US on the conditioned response (CR). The findings confirm that variations in CO2 concentrations produce changes in the CR that are consistent with principles of Pavlovian conditioning. The findings lend additional support to the tenability of a dyspneic suffocation fear theory of panic disorder, a theory that postulates that at least one type of panic attack could be a consequence of Pavlovian conditioning.

Sex differences in relation to conditioned fear-induced enhancement of morphine analgesia

A number of studies have reported that both the immediate and proactive effects of exposure to a shock stressor are less pronounced in female than in male rats. A separate area of research has demonstrated that female rats are less sensitive to the analgesic effects of morphine than males. Experiments from our laboratory, as well as others, have found that exposure to a context associated with shock (i.e., conditioned fear context) at the time of morphine administration, enhances the analgesic effects of morphine. Since previous studies have exclusively employed male rats, the purpose of Experiment 1 was to determine if a sex difference exists to this context conditioned fear-induced enhancement of morphine-induced analgesia. The findings of Experiment 1 showed that females do not appear to exhibit conditioned fear-induced enhancement of morphine analgesia as compared to males. Experiment 2 demonstrated that females exhibited higher levels of conditioned fear-induced enhancement of morphine analgesia during diestrus I than estrous. Experiment 3 demonstrated that females exhibited lower levels of conditioned analgesia compared to males, while both groups exhibited similar freezing levels. The findings of the present experiments suggest that the sex difference observed in Experiment 1 may be due to differences in conditioned analgesia.

Modulation of hypoalgesia by morphine and number of shock trials: Covariation of a measure of context fear and hypoalgesia

In a recent series of studies, we observed that exposure to prolonged foot shock increased hypoalgesia induced by morphine. This increase was observed only when testing was conducted in the presence of shock-associated cues, suggesting that it resulted from context-conditioned fear. However, we do not know whether the extended stressor parameters employed in that study are necessary for an observance of the effect. Therefore, in the present study, we assessed the effect of the number of shock trials (either 0, 20, 100, or 200) on the hypoalgesia observed following morphine administration. In addition, we measured activity as an independent index of context-conditioned fear, because in prior studies there had been no independent behavioral assessment of the conditioning of fear to the context. Although others have shown a covariation of conditioned fear and context-induced hypoalgesia using shock parameters and test paradigms different from our own, we sought to assess whether the same covariation would hold for conditioned fear and the hypoalgesia observed following the administration of morphine. The results showed increased hypoalgesia in all groups exposed to foot shock, demonstrating that prolonged exposure to foot shock is not necessary for an observance of this effect. In addition, the results revealed a linear relationship between number of trials of shock and hypoalgesia, but a U-shaped relationship between trials and activity. The pattern of results is considered in light of Fanselows Perceptual-Defensive-Recuperative model.

Pavlovian aversive context conditioning using carbon dioxide as the unconditional stimulus.

Four experiments were conducted to examine the utility of carbon dioxide (CO2) as an aversive unconditioned stimulus (US) in a Pavlovian context conditioning paradigm. Experiment 1 demonstrated that rats exposed to CO2 in a distinctive context showed elevated levels of freezing relative to controls. Experiment 2 replicated this basic effect with a modified conditioning procedure and additionally demonstrated conditioned analgesia. Experiment 3 demonstrated a positive monotonic relationship between US duration and resistance to extinction of freezing behavior as well as conditioned analgesia. Experiment 4 demonstrated extinction and an extinction-related phenomenon, renewal. These studies clearly demonstrate the utility of CO2 as a Pavlovian US.

Lack of Sex Differences in Anxiety Behaviors During Precipitated Benzodiazepine Withdrawal in Rats

Nonprecipitated benzodiazepine (BZ) withdrawal has been reported to increase anxiety levels in rats. The present experiment determined if gender or hormonal status would modulate putative changes in anxiety-related behaviors during precipitated BZ withdrawal in rats. Intact and gonadectomized male and female rats were treated for 4 weeks with empty or diazepam (DZ)-filled silastic capsules. Animals were injected with the BZ antagonist flumazenil (RO15-1788; 5 mg/kg, i.p.) or vehicle, and immediately placed on the elevated plus-maze. Following the 10-min behavioral test, rats were decapitated and trunk blood was collected to measure corticosterone and gonadal hormone levels. During precipitated BZ withdrawal rats showed significantly decreased percent open-arm time; however, this finding was confounded by a significant decrease in activity levels (e.g., closed-arm entries and total-arm entries). Precipitated BZ withdrawal did not significantly attenuate percent open-arm entries, which factors out drug-induced changes in activity levels, compared to vehicle control animals. Overall, the results of this experiment suggest that precipitated BZ withdrawal does not significantly increase anxiety levels when compared to control animals.

Learned helplessness inducing foot shock can exacerbate morphine responsiveness

Exposure to inescapable tail shock or foot shock has been shown to produce effects on a number of learning tasks. Tail-shock exposure is also known to influence nociception and morphine reactivity. The present series of experiments investigated the effects of foot shock known to induce learned helplessness effects in our laboratory on the subsequent reactivity to morphine. A first set of experiments investigated the hypoalgesic response to a 4 mg/kg dose morphine over 4 consecutive days following exposure to foot shock. Experiment 1A did not reveal an effect of foot shock on morphine-induced hypoalgesia when testing was conducted in a novel context. In Experiment 1B, we observed an increased hypoalgesic response to morphine when testing was conducted in the shock context. The findings of Experiment 1B were replicated in Experiment 2 and extended to assess the contribution of conditioned fear hypoalgesia to these effects. The possible mechanisms responsible for these findings are discussed.