Effects of chronic activity wheel running and imipramine on masculine copulatory behavior after olfactory bulbectomy

We examined the effects of chronic activity wheel running and imipramine administration on appetitive behavior after olfactory bulbectomy (OBX). Male Long-Evans rats were randomly assigned to the following conditions using a 2 x 2 x 2 design: (1) bilateral OBX or sham surgery, (2) voluntary activity wheel running or sedentary home cage, and (3) daily imipramine or saline injections. After 21 days of treatment, animals underwent behavioral testing for copulatory activity and sucrose preference. Bulbectomized animals exhibited decrements in copulatory performance and reductions in sucrose intake compared to sham animals. Within the bulbectomized groups, imipramine-treated rats either did not copulate or had reduced ejaculation frequencies. However, activity wheel running attenuated the copulatory deficits induced by OBX. The findings encourage studies of physical activity and male sexual dysfunction among depressed men being treated by pharmacotherapy.     

Changes in blood pressure and heart rate following bilateral olfactory bulbectomy in rats

Changes in blood pressure and heart rate following bilateral olfactory bulb ablation were investigated in unanesthetized and unrestrained rats with chronic arterial cannula implants. After bilateral olfactory bulbectomy, the rat exhibited a marked increase of emotional responses to given stimuli and a high incidence of muricide. Blood pressure and heart rate at 5 and 10 days after olfactory bulbectomy were significantly lower than in intact rats (rats before bulbectomy) and/or sham operated rats. Marked decreases in blood pressure and heart rate were seen especially in the period when the animals showed exploratory behavior after being transferred to the experimental cage from the home cage. These results suggest that olfactory bulbectomy has a great influence not only on emotionality but also on the central regulatory system of blood pressure in the rat.     

Effect of bilateral olfactory bulbectomy on discrimination avoidance conditioning in rats

Studies were made with the two-way shuttle box method on the acquisition of discrimination avoidance learning by olfactory bulbectomized rats in relation to changes in emotional behavior. Bulbectomized rats showed a marked increase in locomotor activity, with accompanying augmentation of the reactivity and the appearance of muricidal behavior. Initially, the bulbectomized rats showed elevated conditioned avoidance responses to both the CS+ and the CS-. In later stages, there was a continued slow increase in responses to the CS+ accompanied by a decrease in responses to the CS-, until responses to both stimuli were only slightly elevated above the levels shown by control rats. This result suggests that olfactory bulbectomy does not affect discrimination ability itself, but the impairment of discrimination during the initial stages is resulted from hyperemotionality induced by olfactory bulbectomy.     

Role of aversively motivated behavior in the olfactory bulbectomy syndrome.

The aim of the present studies was to determine the extent to which changes in defensive behaviors could account for some of the behavioral effects of bilateral olfactory bulbectomy (OBX) in rats. Four tests of aversively-motivated behavior were conducted in bulbectomized and sham-operated rats: activity in a dimly lit or brightly lit open field, passive avoidance, foot shock-induced freezing, and defensive withdrawal. OBX reduced the duration of immobility in the open field. Bulbectomized rats exhibited less freezing in response to foot shock than sham-operated rats. In the defensive-withdrawal test, bulbectomized rats made more transitions into and spent less time inside the covered enclosure than sham-operated rats. The experiments thus reveal two novel paradigms for assessing the behavioral effects of OBX. The results also suggest that deficits in aversively-motivated behavior, specifically defensive freezing, may comprehensively explain the putative "hyperactivity" and "passive-avoidance learning deficits" widely associated with the OBX behavioral syndrome.     

Olfactory bulbectomy in mice induces alterations in exploratory behavior

The olfactory bulbectomy syndrome is thought to represent a rodent model for psychomotor agitated depression. While this model has been extensively characterized in rats, fewer studies have been conducted with mice. Therefore, the present study aimed at extending the characterization of the OBX-induced behavioral syndrome in mice, using tests like open field, novel object exploration, novel cage and T-maze learning. OBX mice exhibited hyperactivity in a brightly illuminated open field, and also in a novel home cage as well as in the T-maze. Furthermore, OBX mice demonstrated increased exploratory behavior in the novel object test and in the T-maze. The complex alterations described here with respect to locomotion and exploration are robust and can be achieved by relatively simple test procedures. The extended behavioral characterization of the murine OBX model may contribute in particular to the increasing need to test transgenic mice for the presence of depression-like behaviors.     

Male copulatory behavior and female maternal behavior in neonatally bulbectomized rats

Olfactory bulbs were destroyed in six-day-old male and female rats, and anosmia was confirmed in adulthood. Bulbectomized males initiated apparently normal sexual activity at the same age as did control animals. As virgins, bulbectomized females did not injure foster young; when allowed to mate and deliver young, they displayed apparently normal maternal behavior. It was concluded that neonatal olfactory bulbectomy does not preclude the development of essentially normal copulatory behavior in the male rat and maternal behavior in the female rat.     

Effects of olfactory bulbectomy and domicile on stress-induced corticosterone release in the rat.

Following bilateral olfactory bulbectomy or sham surgery, rats were housed either in groups of 5 or individually. After 10 days the animals were sacrificed and plasma corticosterone levels were measured. Half the animals in each group received predictable stress in the form of footshock immediately prior to exsanguination. Two levels of corticosterone elevation were noted, an intermediate level (circa 40 μg/100 ml plasma), in bulbectomised animals without stress and in sham operated animals with stress. Extreme corticosterone elevation (circa 80 μg/100 ml plasma) occurred in bulbectomised rats with stress. The type of housing had no effect on corticosterone elevation. The results are discussed in terms of a non olfactory function for the olfactory bulb, and the role of corticosterone and ACTH in acquisition learning.     

Sexual impairment of inexperienced male rats following pre- and postpuberal olfactory bulbectomy.

Olfactory bulbectomy of sexually inexperienced male rats prevented the occurrence of sexual behavior in most of the operated animals. This effect was observed in rats bulbectomized before puberty as well as after puberty. Bulbectomized rats which had heterosexual experience prior to the operation did not deviate from intact rats. Males living in cohabition with intact males prior to the operation showed only minor deficits in their mating performances. It was concluded that the olfactory lobe while of only minor importance for maintenance of mating once sexual behavior has been initiated, plays an important role in initiation of sexual behavior of the male rat. Furthermore, since treatment with testosterone of prepuberally bulbectomized rats did not stimulate the animals to sexual activity it was concluded that the sexual impairment following bulbectomy of isolated males presumably is not due to an impaired production of gonadal secretions.     

The effects of olfactory bulbectomy and chronic psychosocial stress on serum glucocorticoids and sexual behavior in female rats.

The effects of olfactory bulb removal (OBX) and chronic psychosocial stress on serum glucocorticoids and sexual behavior were assessed in female rats primed with a subthreshold level of estradiol (E2). Ovariectomized females underwent either OBX or sham surgery. Half of the OBX and half of the sham animals were exposed to chronic psychosocial stress (crowding, strobe light, and intermittent noise) for one-half h per day for 27 days. On day 22, three blood samples were collected from each animal for serum corticosterone analysis; 1) before, 2) during, and 3) after acute stress (one-half h restraint). On day 28, females were exposed to either 4 or 24 h of E2. On day 29, all females received 500 micrograms progesterone (P) 4 h prior to sexual behavior testing. Sexual receptivity was measured using the lordosis quotient (LQ = [number of lordotic responses/10 mounts] x 100). Sexual proceptivity (dart and ear wiggling sequences) and rejection (number of nonlordotic responses to mount attempts) were measured throughout a 10-min test period. Results showed that chronic psychosocial stress dramatically increased sexual receptivity and proceptivity while decreasing sexual rejection in female rats primed with a subthreshold level of E2. Chronic psychosocial stress also elevated serum glucocorticoids and significantly exacerbated the glucocorticoid response to acute stress. OBX, while increasing sexual receptivity and decreasing sexual rejection, had no effect on serum glucocorticoids. These findings support the hypothesis that OBX potentiates female sexual behavior directly through an increase in neural sensitivity to E2. In contrast, the enhancing effect of chronic psychosocial stress on female sexual behavior may result from alterations of the hypothalamo-pituitary-adrenal system.     

Dopamine overflow is increased in olfactory bulbectomized rats: an in vivo microdialysis study

Olfactory bulbectomy (OBX) in rats produces behavioral, physiological, and neurochemical changes that resemble symptoms of depression in humans. The procedure thus serves as a rodent model of affective disorder. Many of the behavioral effects of OBX resemble psychomotor agitation. The possible role of dysregulation of ventral striatal dopamine (DA) systems in this phenomenon was investigated. Basal levels of DA, norepinephrine (NE), homovanillic acid, dihydroxyphenylacetic acid, and 5-hydroxyindoleacetic acid were examined in the striatum of OBX and sham-operated controls using in vivo microdialysis. OBX rats exhibited significantly higher basal DA levels (192%) and lower NE levels (12%) than sham-operated controls. Locomotor activity in response to novelty and footshock stress was elevated in OBX rats. The finding of higher DA levels in striatum may explain this "agitation-like" behavior, a commonly observed phenomenon in the OBX model.     

Prenatal stress alters cardiovascular responses in adult rats

Environmental factors in early life are clearly established risk factors for cardiovascular disease in later life. Most studies have focused on nutritional programming and analysed basal cardiovascular parameters rather than responses. In the present study we have investigated whether prenatal stress has long-term effects on cardiovascular responses in adult offspring. Female pregnant Sprague-Dawley rats were subjected to stress three times daily from day 15 to day 21 of gestation. Litters from stressed and control females were cross-fostered at birth to control for mothering effects. When the offspring were 6 months old, blood pressure was measured in the conscious rats through implanted catheters at rest, during restraint stress and during recovery. Basal haemodynamic parameters were similar in the different groups but the pattern of cardiovascular responses during stress and recovery differed markedly between prenatally stressed (PS) and control animals. PS rats had higher and longer-lasting systolic arterial pressure elevations to restraint stress than control animals. They also showed elevated systolic and diastolic blood pressure values during the recovery phase. PS rats demonstrated a greater increase in blood pressure variability compared with control animals during exposure to restraint stress, and showed more prolonged heart rate responses to acute stress and delayed recovery than controls. There was no effect of prenatal stress on baroreflex regulation of heart rate. PS females showed a greater increase in systolic arterial pressure and blood pressure variability and delayed heart rate recovery following return to the home cage then did PS males. These findings demonstrate for the first time that prenatal stress can induce long-term, sex-related changes in the sensitivity of the cardiovascular system to subsequent stress.     

Changes in amygdaloid afterdischarges and kindling effect following olfactory bulbectomy in the rat

Changes in the thresholds for afterdischarges and the formation of kindling effect in the medial amygdala following olfactory bulbectomy were investigated in the rat with chronic electrode implants. The threshold for afterdischarges in the amygdala of the olfactory bulbectomized rat (OB rat) was significantly decreased on day 4 after olfactory bulbectomy, however, no significant difference was found between OB and sham operated rats on days 7, 14 and 21 since the threshold in the sham group was also decreased at these periods after the surgery. The formation of kindling effect was remarkably accelerated in the OB rats. In this case, the number of days required to reach the stage 1 (Racine's classification) was significantly shortened. These results suggest that the activity of the medial amygdaloid nucleus is increased following olfactory bulbectomy.     

Unilateral but not bilateral olfactory bulbectomy inhibits body weight gain in hamsters.

The relation of the olfactory bulbs and photoperiod to the regulation of body weight was studied in male golden hamsters. Animals underwent sham operation, bilateral olfactory bulbectomy, or unilateral bulbectomy. They were left on long photoperiod for 5 weeks and then were transferred to short photoperiod for 11 weeks. The unilaterally olfactory bulbectomized hamsters gained less weight on long or short photoperiod than the sham operated group, while the bilaterally bulbectomized hamsters gained at least as much weight as the sham group. Thus, we report the novel finding that unilateral but not bilateral olfactory bulbectomy reduces body weight gain in male golden hamsters.     

Olfactory bulbectomy induces neuronal rearrangement in the entorhinal cortex in the rat

In humans, depression has been associated with disturbances in olfactory circuitry. Symptoms of depression can be mimicked in animals after olfactory bulbectomy (OBX). Animal models of depression-like behavior produce similar neuronal rearrangements in various brain regions as seen in patients affected by depression. We have recently observed that OBX produces neuronal hypotrophy in the piriform cortex (PirC) and CA1 hippocampus as well as decreased adult cell proliferation in the dentate gyrus (DG) of hippocampus. Thus we further evaluated the effects of OBX in neuronal arborization and spine density in brain regions involved in the control of circadian circle, emotion and memory processing such as the prefrontal cortex (PFC), nucleus accumbens (NAcc), infralimbic cortex (ILC), orbitolateral cortex (OLC) and entorhinal cortex (EC). Our present results show that along with severe behavioral deficits observed in these animals, OBX considerably decreased dendritic branching and the total dendritic length in the EC, a major interface of the hippocampus and neocortical regions. The remaining cortices and NAcc were not affected by OBX. Thus, we propose that the lack of input from the olfactory bulbs resulted in serial neuronal rearrangements in the PirC, EC, and hippocampus leading, at least partially, to behavioral deficits in emotion and memory processes.     

Corticosterone, choline acetyltransferase and noradrenaline levels in olfactory bulbectomized rats in relation to changes in passive avoidance acquisition and open field activity

Consequences of olfactory bulbectomy in two behavioural situations, passive avoidance acquisition and activity in a brightly lit open field, were measured in the same animals for which data on four biochemical measures were also obtained. The biochemical measurements were on plasma corticosterone levels, noradrenaline (NA) levels in the midbrain and amygdala + pyriform cortex and the choline acetyltransferase (CAT) levels in the olfactory tubercle. Experimental variation in age groups of rats (7 weeks and 3 months) and in post-bulbectomy periods (1, 2 and 4 weeks) was made. The deficit in passive avoidance as a consequence of olfactory bulbectomy was evident in all groups of young animals and in older animals one and two weeks post-bulbectomy but not in older animals four weeks after bulbectomy. An increase in open field activity was similarly observed in all groups except in the older animals four weeks after bulbectomy. In contrast to reports by other investigators the basal plasma corticosterone levels were not increased in bulbectomized animals nor did we observe any diminution of NA levels in the amygdala (+ pyriform cortex). CAT levels were slightly increased in older animals two weeks after bulbectomy. The absence of a change in the plasma corticosterone level after bulbectomy is discussed in relation to the notion that the olfactory bulbectomized rat is in some way relevant as a test model for predicting efficacy of potential antidepressant drugs.     

Central serotonin depletion modulates the behavioural, endocrine and physiological responses to repeated social stress and subsequent c-fos expression in the brains of male rats.

Intraspecific confrontation has been used to study effect of depleting central serotonin on the adaptation of male rats to repeated social stress (social defeat). Four groups of adult male rats were used (serotonin depletion/sham: stressed; serotonin depletion/sham: non-stressed). Central serotonin was reduced (by 59-97%) by a single infusion of the neurotoxin 5,7-dihydroxtryptamine (150 microg) into the cerebral ventricles; levels of dopamine and noradrenaline were unaltered (rats received appropriate uptake blockers prior to neurotoxic infusions). Sham-operated animals received solute only. Rats were then either exposed daily for 10 days to a second larger aggressive male in the latter's home cage, or simply transferred to an empty cage (control procedure). Rats with reduced serotonin failed to show the increased freezing behaviour during the pre-defeat phase of the social interaction test characteristic of sham animals. There was no change in the residents' behaviour. Core temperature increased during aggressive interaction in sham rats, and this did not adapt with repeated stress. By contrast, stress-induced hyperthermia was accentuated in serotonin-reduced rats as the number of defeat sessions increased. Basal core temperature was unaffected by serotonin depletion. Heart rate increased during social defeat, but this did not adapt with repeated stress; serotonin depletion had no effect on this cardiovascular response. Basal corticosterone was increased in serotonin-depleted rats, but the progressive reduction in stress response over days was not altered. C-fos expression in the brain was not altered in control (non-stressed) rats by serotonin reduction in the areas examined, but there was increased expression after repeated social stress in the medial amygdala of 5-HT depleted rats. These experiments show that reduction of serotonin alters responses to repeated social stress in male rats, and suggests a role for serotonin in the adaptive process.     

The autonomic stress-induced hyperthermia response is not enhanced by several anxiogenic drugs

While anxiety models are often based on locomotor activity responses, the stress-induced hyperthermia (SIH) paradigm uses the autonomic stress response by measuring body temperature. The effects of putative anxiogenic compounds in the SIH paradigm are inconclusive in mice and have not been examined in rats. Furthermore, it has been suggested that drug-induced effects on body temperature could be dependent on locomotor activity levels. Therefore, the effects of three anxiogenic substances, yohimbine (an α₂ receptor antagonist), mCPP (a 5HT_2C receptor agonist) and FG-7142 (a GABA_A receptor inverse agonist acting at the benzodiazepine site) on the stress-induced body temperature and locomotor activity response were studied in rats using novel cage stress. All anxiogenic compounds resulted in hypothermia. In contrast, FG-7142 and yohimbine increased locomotor activity levels, whereas mCPP reduced locomotor activity levels. The lack of an increased body temperature response of anxiogenic compounds indicates that the anxiogenic capacity of a drug does not necessarily yield increased autonomic stress responsivity. Moreover, the present study shows that a drug-induced decreased body temperature can be accompanied by increased locomotor activity, suggesting that both parameters represent independent parameters of the stress response.     

Experimental and husbandry procedures as potential modifiers of the results of phenotyping tests

To maximize the sensitivity of detecting affects of genetic variants in mice, variables have been minimized through the use of inbred mouse lines, by eliminating infectious organisms and controlling environmental variables. However, the impact of standard animal husbandry and experimental procedures on the validity of experimental data is under appreciated. In this study we monitored the impact of these procedures by using parameters that reflect stress and physiological responses to it. Short-term measures included telemetered heart rate and systolic arterial pressure, core body temperature and blood glucose, while longer-term parameters were assessed such as body weight. Male and female C57BL6/NTac mice were subjected to a range of stressors with different perceived severities ranging from repeated blood glucose and core temperature measurement procedures, intra-peritoneal injection and overnight fasting to cage transport and cage changing.Our studies reveal that common husbandry and experimental procedures significantly influence mouse physiology and behaviour. Systolic arterial pressure, heart rate, locomotor activity, core temperature and blood glucose were elevated in response to a range of experimental procedures. Differences between sexes were evident, female mice displayed more sustained cardiovascular responses and locomotor activity than male mice. These results have important implications for the design and implementation of multiple component experiments where the lasting effects of stress from previous tests may modify the outcomes of subsequent ones.     

Impaired temperature regulation in rats after anosmia induced peripherally or centrally

Bilateral olfactory bulbectomy or destruction of the olfactory epithelium of rats resulted in elevated body temperature in room temperature, and lowered water/food ratio in 30 degrees ambient temperature. The results suggest the involvement of the olfactory system in the thermoregulation.     

A study of VitalView for behavioural and physiological monitoring in laboratory rats

We describe the use of a commercially available telemetry and data acquisition system to record heart rate, body temperature and activity of freely behaving rats with transmitters that operate without batteries (transponders). The system uses PDT 4000HR E-Mitters (Mini Mitter, OR, USA) to acquire animal temperature, heart rate and motor activity data. E-Mitters obtain power from a radiofrequency field produced by an ER-4000 energizer/receiver so that transponders can collect data on heart rate, body temperature and gross motor activity. ER-4000 energizers/receivers are designed to be placed below the animals' cage. Data output from receivers is managed by a Windows PC-based data acquisition system, VitalView. In this study, we report that a good correlation exists between VitalView and Powerlab for the determination of heart rate and between intra-abdominal (telemetric) and colonic body temperature (rectal digital thermometer) in rats. Assessment of this system by using agents that have well-documented effects on heart rate, body temperature and locomotor activity have also been determined. An additional feature of VitalView is the incorporation of behavioural inputs (feeding monitors to monitor duration and frequency of feeding and a lickometer to monitor drinking bouts) into the data acquisition system designed primarily to acquire data from the implanted transponders. Circadian rhythms for all parameters were established in rats with E-Mitters implanted. VitalView may be used for the determination of multiple parameters in freely behaving animals using transponders, which operate without batteries. This capability is unique in its field and represents a recent advance in biotelemetric monitoring of laboratory animals.