Olfactory bulbectomy induces rapid and stable changes in basal and stress-induced locomotor activity,heart rate and body temperature responses in the home cage

Background: Olfactory bulbectomy (OBX) in rats causes several behavioral and neurochemical changes. However, the extent and onset of physiological and behavioral changes induced after bulbectomy have been little examined. Methods: Male Sprague–Dawley rats received telemetric implants. Before and immediately after OBX surgery, basal and stress-induced heart rate, body temperature, and locomotor activity were measured in the home cage in sham (n=9) and OBX animals (n=11). Stress was induced using novel cage stress or witness stress. Results: Bulbectomized animals differed physiologically and behaviorally from shams. Nocturnally, OBX animals were significantly more active compared with shams, had a higher core body temperature and displayed a decreased heart rate variability. During the light period, OBX animals had a significantly lower basal heart rate and a reduced heart rate variability. These effects became apparent 2–3 days after OBX surgery, and were stable over time. After witness stress, OBX animals showed smaller autonomic (body temperature and heart rate) responses compared with shams, but showed no difference in locomotor responses. In contrast, novel cage stress led to increased locomotor responses in OBX rats compared with sham rats, while no differences were found in autonomic responses. Conclusion: Removal of the olfactory bulbs results in rapid, stable and persistent changes in basal locomotor activity, body temperature, heart rate and heart rate variability. Although the sleep–wake cycle of these parameters is not altered, increases in circadian amplitude are apparent within 3 days after surgery. This indicates that physiological changes in the OBX rat are the immediate result of olfactory bulb removal. Further, stress responsivity in OBX rats depends on stressor intensity. Bulbectomized rats display smaller temperature and heart rate responses to less intense witness stress compared with sham rats. Increased locomotor responses to more intense novel cage stress are present in the home cage as well as the open field. The present study shows that olfactory bulbectomy has rapid and persistent influence on basal and stress-induced physiological parameters.     

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.     

Olfactory system involvement in the recovery of feeding and drinking after lateral hypothalamic damage in the rat.

In rats, large lesions of the far lateral hypothalamus produce a familiar syndrome of behavioral deficits which include the absence of feeding and drinking. If tube fed and offered palatable food LH lesioned rats gradually recover and eventually become able to maintain a stable body weight on dry food and water. In this series of experiments we show that although bilateral removal of the olfactory bulbs does not affect a rat's ability to feed and drink to maintain a normal body weight (Experiment 1), bulbectomy performed about one month prior to lateral hypothalamic lesioning can seriously jeopardize the recovery of feeding and drinking after lateral hypothalamic damage (Experiment 2). Additionally (Experiment 3), in some recovered lateral hypothalamically lesioned rats bulbectomy is followed by a period of persistent adipsia and anorexia from which recovery nevertheless occurs. Thus, although the olfactory bulbs may make a contribution to the maintenance of drinking and feeding in the recovered LH rat, this contribution is apparently not an indispensible one.     

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.     

Olfactory bulbectomy depresses ultrasound production and scent marking by female hamsters

Female hamsters were observed for rates of ultrasound production, vaginal marking and flank marking before and after bilateral olfactory bulbectomy, unilateral bulbectomy or sham operations. Unilaterally bulbectomized and sham-operated subjects maintained preoperative levels of each behavior. In contrast, females with bilateral olfactory bulbectomies showed consistent decreases in rates of vocalization and scent marking. These results emphasize the dependence on an intact olfactory system of several courtship or agonistic behaviors that affect hamster reproduction by functioning as noncontact communications. Together with the results of previous studies suggesting that lordosis by female hamsters is unaffected by olfactory bulbectomy, these data also suggest differences in the brain and sensory mechanisms that control distinct types of reproductive behavior.     

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.     

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.     

Activity-induced manganese-dependent functional MRI of the rat visual cortex following intranasal manganese chloride administration

Manganese-enhanced MRI (MEMRI) of the brain requires delivery of manganese into the target brain regions. It was previously shown that, following intranasal application, ongoing olfactory stimulation facilitates manganese transport along the olfactory nerve into the olfactory bulb, so bypassing the blood–brain barrier (BBB). We report on experiments to evaluate whether visual stimulation can permit manganese transport onwards from the olfactory bulb to the visual cortex. Rats in intact olfactory bulb group were reserved intact olfactory bulb, while those in olfactory bulbectomy group received bilateral bulbectomy. After intranasal MnCl₂ administration, olfactory and visual stimulations were performed on all the animals for a consecutive 20 h. The visual cortex was then examined using MEMRI. Enhanced imaging on T1WI was noted in the visual cortex of the intact olfactory bulb group. Image subtraction revealed that the signal intensity in the visual cortex of the intact olfactory bulb group was significantly higher than that of olfactory bulbectomy group. Volume of interest (VOI) analysis also showed that normalized intensities in the visual cortex of the intact olfactory bulb group were significantly higher as compared with those of the olfactory bulbectomy group. Inductively coupled plasma mass spectrometry (ICP-MS) confirmed that the manganese content in the visual cortex of the intact olfactory bulb group was increased in comparison with that of the olfactory bulbectomy group. These findings indicate that activity-induced manganese-dependent functional MRI (AIM fMRI) of the rat visual cortex can be performed following intranasal administration of manganese and demonstrate that manganese can migrate from the olfactory bulb to the visual cortex.     

Tail pinch induces sexual behavior in olfactory bulbectomized male rats

Adult male Long-Evans rats were subjected to bilateral olfactory bulbectomy, sham surgery or no treatment. Of 34 bulbectomized rats, 24 failed to ejaculate on either of 2 tests with a primed ovariectomized female. All control animals exhibited normal sexual behavior, and 10 bulbectomized animals ejaculated at least once during the 2 tests. Later histological examination revealed a relationship between size of lesion and extent of behavioral deficits. After a third test, 16 nonejaculatory animals were subjected to a tail pinch (TP) procedure, immediately followed by a fourth test. The remaining 8 nonejaculatory animals were tested similarly, but without tail pinch. Ten of the 16 tail pinch animals showed complete sexual behavior on the first test, while 2 additional animals began to copulate after a second TP procedure 4 days later. Only 1 of the 8 animals not receiving TP ejaculated on these tests. Thus, TP applied shortly before sexual behavior tests can induce copulation in some males whose behavior had been disrupted by olfactory bulbectomy.     

The area postrema and control of arterial blood pressure; Absence of hypertension after excision of the area postrema in rats

Summary Acute surgical excision of the area postrema (AP) in the rat failed to affect arterial blood pressure or heart rate.There was no effect on cardiovascular reflex responses during diving or on the heart rate responses to acute decreases or increases of blood pressure caused by bradykinin or angiotensin, respectively. Electrolytic lesions of the AP in acute experiments caused variable damage to the nucleus tractus solitarii (NTS). In these rats large variations in blood pressure occurred. Excision of the AP in a chronic experiment failed to change blood pressure, heart rate, water intake or plasma renin activity. In contrast, bilateral electrolytic lesions of the NTS at the level of the AP caused a severe acute hypertension and completely blocked cardiovascular reflex responses. Hypertension also existed in rats with NTS lesions studied for a longer period of time.These experiments failed to confirm the hypothesis that the AP exerts a tonic inhibitory control of basal blood pressure. Hypertension previously reported after ablation of the AP may be explained by damage to the NTS.     

Effects of olfactory bulb removal and flank shock on copulation in male rats

Sexually experienced male rats received bilateral olfactory bulbectomy (BOB, N=9), unilateral olfactory bulbectomy (UOB, N=7) or a sham operation (Sham, N=8). Copulation was severely disrupted in BOB males, whereas only minimal effects on copulation were observed in UOB males. Penile reflexes were not affected by either operation. Intermittent flank shock stimulated copulation in 5/6 BOB males. The BOB males that ejaculated had longer intromission latencies and longer postejaculatory intromission intervals than did Sham males receiving a similar schedule of shocks. Even so, all BOB males that ejaculated with flank shock also resumed copulation and ejaculated a second time without further shocks. Three BOB males that ejaculated with flank shock were tested the following week without shock, and none of these males copulated. These results strengthen previous conclusions that the olfactory bulbs are critical for the initiation and maintenance of sexual arousal in male rats.     

Bulbectomy and sensitivity to estrogen: Anatomical and functional specificity

In order to clarify the influence of the olfactory system on female sexual behavior, ovariectomized rats were given sham operations (SHAM), total bilateral olfactory bulbectomy (TBULBX), partial bulbectomy (PBULBX), anterior olfactory nucleus lesions (AON) or accessory olfactory bulb lesions (AOB), and tested for lordosis behavior. Only TBULBX resulted in increased sensitivity to estradiol benzoate (EB) in that lordosis quotients (LQ) were increased and rejection behavior decreased following administration of 2, 4 or 8 μg EB/kg/day for 3 days. Only TBULBX group rats were anosmic on 2 postoperative tests. TBULBX group rats showed very mild hyperresponsiveness on an emotionality test. Effects of TBULBX on LQ are not due to general sensory hyperresponsiveness or EB-induced hyperresponsiveness since no differences in the quality of lordosis occurred, and no differences occurred in latency to paw-lift on hot plate tests with or without EB. Heightened EB sensitivity in the TBULBX group is not due to adrenal steroids since following adrenalectomy and 8 μg EB/kg treatment, TBULBX group LQ scores were still elevated relative to those of SHAM controls. The LQ scores of PBULBX group rats were intermediate to those of SHAM and TBULBX group rats. Bulbectomy-induced alterations in sensitivity to EB as measured by the LQ do not appear to be due to alterations in “arousal” mechanisms in general. While deficits in olfactory perception might exacerbate the effect, it is unlikely that anosmia per se is sufficient to induce major alterations in the degree of sexual receptivity following EB. The magnitude of behavioral effects of bulbectomy on EB sensitivity may be related, to some extent, to the amount of bulb tissue removed. It is possible that bulbectomy may enhance behavioral sensitivity to EB by disrupting biochemical responses to EB in limbic system structures which normally exert an inhibitory influence over sexual receptivity.     

Brain transplants enhance rather than reduce the impairment of spatial memory and olfaction in bulbectomized rats

The possibility to compensate the loss of olfactory and non-olfactory functions due to removal of the olfactory bulb by embryonal brain grafts was investigated in adult rats. Spatial working memory was examined in an 8-arm radial water maze task 6 weeks after bulbectomy. During 15 daily trials, performance gradually improved in bulbectomized controls (n = 10) and in rats with olfactory bulb transplants (n = 9), but did not attain that of intact controls (n = 10). No improvement was observed in the rats with substantia nigra grafts (n = 8). Eleven weeks after bulbectomy, the same rats were tested in the water tank navigation task. The performance improved during ten 12-trial sessions in bulbectomized rats less than in intact controls, but more than in the transplanted rats. The olfactory food retrieval test performed 14 weeks after bulbectomy revealed almost full recovery of smell in bulbectomized rats, but not in the transplanted animals. It is concluded that the spatial memory deficit is probably due to bulbectomy-induced interference with septohippocampal function which is not alleviated, but rather enhanced by transplantation. The results suggest that the effect of brain grafting is not always beneficial.     

Differential effects of medial, central and basolateral amygdaloid lesions on four models of experimentally-induced aggression in rats

To clarify whether various nuclei of the amygdaloid complex play different roles in aggressive behavior including muricide, 4 types of aggression were experimentally induced in rats. These include olfactory bulbectomy (OB rats), midbrain raphe lesions (Raphe rats), administration of delta 9-tetrahydrocannabinol (THC rats) and long-term isolation (Iso rats). Rats which exhibited muricide following these treatments were subjected to bilateral lesions of either the medial (AME), central (ACE) or basolateral (ABL) amygdaloid nuclei. Both muricide and hyperemotionality in the OB rat were markedly inhibited by AME lesions. Those of the Iso and THC rats were moderately inhibited. However, in the Raphe rat, aggressive behavior was not inhibited by AME lesions. Furthermore, ACE or ABL lesions caused no significant changes in all 4 models of aggression. These results suggest that the AME plays a facilitatory role in aggression of OB, Iso and THC rats, but aggression in Raphe rat is independent of amygdaloid activity.     

Influence of isolation and training on fighting in mice with olfactory bulb lesions

Isolation-induced aggressive mice receiving total bilateral bulbectomy failed to fight after chronic training. Animals receiving sub-total olfactory bulbectomy were capable of being trained to attack but the latency to attack was increased. When mice were bulbectomized before being isolated they were incapable of attack regardless of training or completeness of the lesion. These results indicated that isolation influences aggressive behavior in bilaterally bulbectomized mice.     

Olfactory bulb removal affects partner preference development and estrus induction in female prairie voles.

In female prairie voles (Microtus ochrogaster) bilateral olfactory bulbectomy reduced affiliative behavior, as measured by social contact, and prevented the formation of partner preferences. Unilateral olfactory bulb removal did not significantly influence affiliative behavior, but did inhibit partner preferences. Bilateral, but not unilateral, bulbectomy significantly reduced the proportion of females exhibiting behavioral estrus following male exposure. In contrast to affiliative and sexual behavior, parental behavior was not significantly affected by either bilateral or unilateral olfactory bulbectomy. These results suggest that divergent sensory-neural pathways underlie social, sexual, and parental behaviors in this species.     

Effects of ovarian hormones on sexual receptivity, proceptivity, and motivation in olfactory bulbectomized female rats.

The purpose of the present experiment was to assess the effect of olfactory bulbectomy and ovarian hormones on female sexual motivation. Ovariectomized female rats underwent either bilateral bulbectomy or sham surgery. Females received one of four subthreshold hormone treatments: 0% estradiol (E2) plus 500 micrograms progesterone (P), 100% E2 alone, 10% E2 plus 500 micrograms P, or 100% E2 plus 500 micrograms P. Sexual motivation (as indicated by a female's preference for a sexually active male over a castrated male) and proceptivity (dart and ear wiggling sequences) were measured in a three compartment partner preference apparatus. Sexual receptivity (lordosis) was measured separately in a glass arena with a sexually active male. Results showed that olfactory bulb removal facilitates sexual receptivity and proceptivity in females exposed to 10% or 100% E2 in combination with 500 micrograms P. In contrast, sexual motivation was only demonstrated by olfactory bulbectomized females which received 100% E2 in combination with 500 micrograms P. These findings support the hypothesis that olfactory bulbectomy induces a behavioral hypersensitivity to estrogen, and suggest that sexual motivation is an estrogen-mediated response which requires a higher level of estrogen stimulation than sexual receptivity and proceptivity.     

Olfactory bulbectomy: influences on maternal behavior in primiparous and multiparous rats

Bilateral olfactory bulb ablation resulted in deficits in several components of maternal behavior in lactating, primiparous female rats. These females frequently cannibalized pups shortly after parturition. Furthermore, bilaterally bulbectomized, primiparous females spent less time nursing their pups and retrieved fewer pups than unilaterally bulbectomized or sham-operated females. In contrast to the situation in primiparous females, in multiparous females bilateral bulbectomy failed to influence any parameter of maternal behavior.     

Increased cardiac contractility in acute uremia: interrelationships with hypertension.

To study the effects of acute uremia on the inotropic state of the rat heart, we subjected rats to bilateral nephrectomy and studied their hearts in the open chest 24 h later. Uremic rats had significantly higher systolic blood pressure than sham-operated animals. Left ventricular systolic pressure and maximum dP/dt, both during ejection and isovolumic contrations, were higher for any given end-diastolic pressure in hearts of uremic rats than in sham-operated animals. This difference in performance charcteristics was not abolished by doses of propranolol that blocked the heart rate response to isoproterenol. The administration of phenoxybenzamine during the 24 h of uremia abolished the blood pressure rise in uremic rats, but the increased contractile state persisted. Treatment of sham-operated animals with methoxamine to produce the same course of blood pressure as observed in uremic rats was also associated with an increased inotropic state. These results indicate that in the rat, acute uremia is associated with an increased inotropic state that is not mediated by beta-adrenergic mechanisms. The systolic hypertension of acute uremia is not the major cause of the increased contractility, although systolic hypertension without uremia can mimic the performance characteristics found in hearts of uremic rats.     

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.