Lordosis behavior in intact male rats: effects of hormonal treatment and/or manipulation of the olfactory system

The aim of this study was to investigate the olfactory mechanisms regulating the display of lordosis behavior in intact Wistar male rats bred in our colony. Gonadally intact males show a low capacity to respond by lordosis to male mounts and were insensitive to manipulations of the olfactory system (exposure to the odor of male urine or accessory bulb removal (AOBR)) which have been previously shown to facilitate the display of lordosis behavior in orchidectomized animals primed with ovarian hormones. Treatment with either estradiol benzoate (EB) or EB and progesterone (P) consecutively did not render these gonadally intact animals sensitive to the effects of AOBR. By contrast exposure to male urine was capable of facilitating the display of lordosis behavior in intact male rats given EB + P consecutively. These results are discussed in the light of previous findings showing that (1) two inhibitory structures, the accessory olfactory bulb and the septal and preoptic areas, are involved in the control of lordosis behavior in the male rat; (2) the effects of olfactory cues on the display of lordosis behavior are dependent on the action of both EB and P in orchidectomized animals.     

Accessory olfactory bulb lesions and lordosis behavior in the male rat feminized with ovarian hormones

Orchidectomized rats were given estrogen and progesterone and tested for feminine behavior in the presence of a mounting male after accessory olfactory bulb removal (AOBR). Complete AOBR caused a rise in the number of estrogen-progesterone-treated male rats responding by lordosis behavior to male mounts as compared to controls and sham-operated animals. By contrast, LQ scores did not appear to differ in these three groups of animals. The results are discussed in terms of involvement of the main and the accessory olfactory systems in the regulation of feminine behavior in the male rat.     

Changes in estrogen receptors in the mediobasal hypothalamus mediate the facilitory effects exerted by the male's olfactory cues and progesterone on feminine behavior in the male rat

The purpose of this study was to determine whether facilitory effects exerted by olfactory cues on lordosis behavior in the male rat involved changes in estradiol receptors at the hypothalamic level. Male rats were orchidectomized as adults. They were given either 25 micrograms estradiol benzoate (EB) alone or 25 micrograms EB and 100 micrograms progesterone (P) sequentially and exposed or not to the odor of male urine. Some of them were tested for lordosis behavior at 8 h after P. The other ones were killed 4 h after P and used for estradiol (E2) and P receptor assay in mediobasal hypothalamus (MBH). Olfactory cues were shown to increase the number of E2 receptors in both the animals given EB or EB + P. Progesterone as such appeared to be capable of increasing the number and the rate of occupancy of E2 receptors. A population of constitutive and estrogen-inducible P receptors was detected in the MBH. Since only the animals given EB + P were shown to be sensible to the facilitory effects of male urine on lordosis behavior, it may be assumed that E2 and P on one hand and olfactory cues on the other exert cumulative effects at the level of the MBH and that both a high level and a high rate of occupancy of E2 receptors are necessary for the olfactory cues to facilitate the display of lordosis behavior in the male rat.     

Female lordosis pattern in the male rat induced by estrogen and progesterone: effect of interruption of the dorsal inputs to the preoptic area and hypothalamus.

Lordosis behavior was very rare in castrated male rats which had been pretreated with 50 mug estradiol benzoate (EB) for successive 2 days and 1 mg progesterone (P) 6-8 hr prior to testing on the third day. Only one out of 8 rats displayed lordosis in response to mounts by the sexually matured males. However, the occurrence of lordosis behavior was markedly increased in similarly treated castrated males in which the dorsal afferents to the preoptic area (POA) and hypothalamus were removed by the surgical cut. Twelve out of 19 rats of the group showed lordosis response. The incidence was less frequent in rats receiving sham deafferentation. These results suggest that the dorsal inputs to the POA and hypothalamus may exert a tonic inhibitory influence on the lordosis mediating system in the male rat.     

gamma-Aminobutyric acid activity in the olfactory bulb of the rat during the sexual cycle and response to olfactory stimuli.

Glutamic acid decarboxylase activity in the main and accessory olfactory bulbs throughout the sexual cycle of the rat was studied. The effect of male pheromonal secretion on enzyme activity during proestrus and estrus day was also tested. The enzyme activity showed circadian rhythm during the estrous cycle. This rhythm was disrupted during diestrus-2 afternoon in the main bulb and came back during proestrus afternoon. A different pattern of enzyme activity was present in the accessory bulb, since the circadian rhythm was altered during proestrus morning, returning during estrus afternoon. Male odor exposition did not change enzyme profile activity during proestrus day and during estrus morning in the main bulb. In contrast, in the accessory bulb the olfactory stimuli induced opposite changes to that found in rats from the vivarium during proestrus. Comparison of enzyme activity in olfactory stimuli-deprived rats with that of pheromone-stimulated rats during proestrus showed that male odor exposure specifically affects accessory bulb enzyme activity. It is concluded that the changes of the olfactory bulb GABAergic system during proestrus and estrus day, or that evoked by odor stimuli, demonstrate the discriminative response of this system between the accessory olfactory bulb and the main olfactory bulb.     

The reversible inhibition of steroid-induced sexual behavior by intracranial cycloheximide

Cycloheximide(Cyclo), an inhibitor of protein synthesis by a direct action on protein synthesis at the ribosomal level, was used to reversibly inhibit estrogen-induced sexual receptivity. Cyclo (100 μg per rat) was infused into the preoptic area(POA) of ovariectomized rats at varying times before, simultaneously with, and after 3 μg of subcutaneous estradiol benzoate (EB). All animals received 0.5 mg progesterone (P) 36 hr after EB, and were tested for sexual receptivity 4–6 hr after P. The females were placed with stud males and a lordosis quotient was computed for each female (lordosis quotient = number of lordosis responses/20 mounts by the male × 100). Females receiving Cyclo 6 hr before, simultaneously with, or 12 hr after EB showed significantly lower levels of sexual receptivity when compared to females receiving Cyclo 36 hr before and 18 and 24 hr after EB. When those animals that showed low levels of sexual behavior after Cyclo infusion were reprimed with EB and P 7 days later and presented with a male they showed high levels of sexual receptivity. Thus, the effect of Cyclo was reversible. Only Cyclo infusions into the POA (bilateral) and third ventricle were effective in suppressing sexual behavior. Caudate nucleus, lateral ventricle, and unilateral POA infusions were without effect.The data presented are in agreement with earlier work that utilized actinomycin D to inhibit steroid-induced sexual behavior. Cyclo was found to be less toxic than actinomycin D. All of the available evidence is consistent with the hypothesis that estrogen stimulates RNA and/or protein synthesis in its facilitation of sexual behavior in the female rat.     

Neural and behavioral changes in rats following continuous exposure to an odor

Summary Continuous exposure of young rats to the almond-like odor of acetophenone or cyclohexanone for up to 4 months, resulted in distinct but similar patterns of degenerating mitral cells in their olfactory bulbs. Rats favored their exposure odor in olfactory preference tests (Fig. 2) and their acuity for it was not altered (Fig. 3). However, they appeared to exhibit a deficit in detecting a similar but novel odor. The results suggest that the remaining normal mitral cells in the bulbs of these animals are those stimulated by the exposure odor. Cells which show signs of degeneration (Fig. 4) may receive little or no input from the periphery. Controls exposed to a similar but non-odorous environment showed evidence of non-selective mitral cell degeneration. In addition they had a lower acuity for acetophenone and cyclohexanone than animals reared in a normal rat colony (Fig. 3). Anatomical and behavioral data from odor exposed and control groups, suggest that partial regeneration of altered mitral cells may have occurred during a 5 month period following exposure. Overall the results provide further evidence for a topographical projection of the olfactory receptor epithelium onto the olfactory bulb and spatial coding of different odors in the bulb.     

Photoperiod Mediated Changes in Olfactory Bulb Neurogenesis and Olfactory Behavior in Male White-Footed Mice (Peromyscus leucopus)

Brain plasticity, in relation to new adult mammalian neurons generated in the subgranular zone of the hippocampus, has been well described. However, the functional outcome of new adult olfactory neurons born in the subventricular zone of the lateral ventricles is not clearly defined, as manipulating neurogenesis through various methods has given inconsistent and conflicting results in lab mice. Several small rodent species, including Peromyscus leucopus, display seasonal (photoperiodic) brain plasticity in brain volume, hippocampal function, and hippocampus-dependent behaviors; plasticity in the olfactory system of photoperiodic rodents remains largely uninvestigated. We exposed adult male P. leucopus to long day lengths (LD) and short day lengths (SD) for 10 to 15 weeks and then examined olfactory bulb cell proliferation and survival using the thymidine analog BrdU, olfactory bulb granule cell morphology using Golgi-Cox staining, and behavioral investigation of same-sex conspecific urine. SD mice did not differ from LD counterparts in granular cell morphology of the dendrites or in dendritic spine density. Although there were no differences due to photoperiod in habituation to water odor, SD mice rapidly habituated to male urine, whereas LD mice did not. In addition, short day induced changes in olfactory behavior were associated with increased neurogenesis in the caudal plexiform and granule cell layers of the olfactory bulb, an area known to preferentially respond to water-soluble odorants. Taken together, these data demonstrate that photoperiod, without altering olfactory bulb neuronal morphology, alters olfactory bulb neurogenesis and olfactory behavior in Peromyscus leucopus.     

Induction of progestin receptors in the mediobasal hypothalamus of gonadally intact male rats primed with estrogen in relation to display of lordosis behavior

The purpose of this study was to determine whether the effects of estrogen on lordosis behavior in the male rat were related to the number of progesterone (P) receptors in the mediobasal hypothalamus (MBH) and/or dependent on blood P concentration. Two groups of gonadally intact male rats were given five successive doses of 1.0 or 2.5 micrograms estradiol benzoate (EB) and tested for lordosis behavior with a male stimulus at the end of the treatment. One month later they were again injected with EB and sacrificed under the same temporal schedule, but they were not tested for lordosis so as to prevent any emotionally stressful effects of intermale cohabitation. The males given 2.5 micrograms EB more frequently displayed lordosis responses to male mounts than those receiving 1 microgram EB, with a parallel increase in the number of MBH P receptors. The total number of MBH P receptors also appeared to be higher in the animals that displayed lordosis responses (lordosis group) than in those which did not (no lordosis group). In contrast, the display of lordosis behavior was negatively correlated with blood P concentration. Comparing MBH P receptors and blood P values in the EB treated and in nonhormonally treated gonadally intact animals which had been selected for either ability or inability to spontaneously display lordosis behavior, we observed that (1) EB was capable of increasing the number of MBH P receptors in the male rat; and (2) in the absence of EB treatment blood P values were higher in the animals showing lordosis than in those which did not. These data are discussed with respect to observations made in castrated male rats and in ovariectomized females.     

Protease-sensitive urinary pheromones induce region-specific Fos-expression in rat accessory olfactory bulb.

Vomeronasal organs of female Wistar rats were exposed with sprayed urine preparations of male Wistar rats prior to sacrifice. Exposure to crude urine and ultrafiltrated urine preparation (<5000 Da) induced significant Fos expression, which is correlated with cellular activity, in the mitral/tufted cell layer of the accessory olfactory bulb (AOB), while exposure to the remaining substances after ultrafiltration (>5000 Da) and control salt solution did not. Exposure to urine preparation treated with papain induced expression of Fos-immunoreactive cells in the rostral region of the AOB, but did not induce such expression in the caudal region. Exposure to urine preparation treated with pronase induced urine-specific Fos immunoreactivity neither in the rostral nor in the caudal region. These results suggest that at least two different peptides carrying pheromonal activities are contained in male Wistar rat urine.     

Mu-, delta-, and kappa-opioid receptor agonists selectively modulate sexual behaviors in the female rat: differential dependence on progesterone.

Previous studies suggested that opioid receptor agonists infused into the lateral ventricles can inhibit (through mu receptors) or facilitate (through delta receptors) the lordosis behavior of ovariectomized (OVX) rats treated with estrogen and a low dose of progesterone. The present study investigated the behavioral and hormonal specificity of those effects using more selective opioid receptor agonists. Sexually experienced OVX rats were implanted stereotaxically with guide cannulae aimed at the right lateral ventricle. One group of rats was treated with estradiol benzoate (EB, 10 micrograms) 48 hr and progesterone (P, 250 micrograms) 4 hr before testing, whereas the other group was treated with EB alone. Rats were infused with different doses of the selective mu-receptor agonist DAMGO, the selective delta-receptor agonist DPDPE, or the selective kappa-receptor agonist U50-488. The females were placed with a sexually vigorous male in a bilevel chamber (Mendelson and Gorzalka, 1987) for three tests of sexual behavior, beginning 15, 30, and 60 min after each infusion. DAMGO reduced lordosis quotients and magnitudes significantly in rats treated with EB and P, but not in rats treated with EB alone. In contrast, DPDPE and U50-488H increased lordosis quotients and magnitudes significantly in both steroid-treatment groups. Surprisingly, measures of proceptivity, rejection responses, and level changes were not affected significantly by mu or kappa agonists, although proceptivity and rejection responses were affected by DPDPE treatment. These results suggest that the effects of lateral ventricular infusions of opioid receptor agonists on the sexual behavior of female rats are relatively specific to lordosis behavior. Moreover, the facilitation of lordosis behavior by delta- or kappa-receptor agonists is independent of progesterone treatment, whereas the inhibitory effect of mu-receptor agonists on lordosis behavior may require the presence of progesterone.     

Heterotypic sexual behavior in male rats after lesions in different amygdaloid nuclei

Previous studies showed that different amygdaloid nuclei are involved in the control of lordosis behavior in female rats. The objective of the present study was to determine whether these nuclei played a role in the control of lordosis behavior in the male rat. Lesions were placed into different amygdaloid nuclei in male rats castrated as adults and primed with ovarian hormones. Lesions in the corticomedial amygdaloid nucleus completely suppressed lordosis behavior as expressed by the number of animals displaying lordosis responses to male mounts. By contrast extended lesions placed into the lateral amygdaloid nucleus (LN) remained without effects. Lesions placed in the very posterior part of the LN produced "hypersexuality" with a rise in the number of animals displaying lordosis responses and high LQ values. Lesions in the anterior part of the LN induced a decrease in the number of animals showing lordosis responses. The amygdala was then concluded to represent a functionally heterogeneous structure with different regions exerting opposite effects on the display of lordosis behavior in the male rat.     

Estrogen and progesterone interactions influencing sexual and social behavior in the brown lemming, Lemmus trimucronatus.

The effects of estrogen and progesterone on the social and sexual behavior of brown lemmings, Lemmus trimucronatus, were investigated. The behavior of hormone-treated and untreated ovariectomized females and sexually vigorous males was observed in six consecutive daily 5-min dyadic encounters. Sexual receptivity, as measured by lordosis, and other social behaviors including nasonasal contact, boxing postures, allogrooming, perineal investigation, and male mounting increased following 48 hr of exposure to daily injections of 0.5 μg estradiol benzoate (EB). Lordosis in EB-primed females was not facilitated or inhibited by short-term (4 hr) exposure to 0.5 mg progesterone (P). Long-term (greater than 24 hr) exposure to P apparently inhibited lordosis and other social behaviors in EB-treated females, although males continued to attempt to mount these females. In EB-treated females a dramatic increase in threat-leaps, directed by the female toward the male, was observed within 4 hr of P injection. Threat-leaps declined when P was withdrawn. Threat-leaps were also observed in ovariectomized females after prolonged exposure to P only (0.5 mg/day). Vaginal perforation and cornification were first apparent 48 hr after EB injection. P-alone treated ovariectomized females also showed vaginal perforation but cornified cells were infrequent and these animals did not show lordosis.     

Chronic exposure of cat odor enhances aggression,urinary attractiveness and sex pheromones of mice

To test whether predator odor exposure negatively affects the behavior of prey, we exposed three groups of male house mice (Mus musculus) to the odors of cat (Felis catus) urine, rabbit (Oryctolagus cuniculus) urine and water (control), respectively, for consecutive 58 days and investigated how the treatments affected the response, aggressiveness, dominance, urinary attractiveness to females and pheromone composition of male mice. Compared to mice exposed to rabbit urine or water, those exposed to cat odor did not show any response habituation to the cat odor and became more aggressive, increased mark urine production and were more attractive to females when the latter were tested with their urine. Furthermore, gas chromatography coupled with mass spectrometry analysis revealed coincident elevations of the well-known male pheromones, E,E-α-farnesene, E-β-farnesene, R,R-dehydro-exo-brevicomin or S-2-sec-butyl-dihydrothiazole. In addition, rabbit urine exposure increased urinary attractiveness to females and pheromonal levels of the males in comparison with the mice exposed to water. This could be related to olfactory enrichment of heterospecific chemosignals, suggesting that predator odors were more beneficial. In light of these anti-intuitional findings in the chemical interaction between cats and mice, we conclude that predator odor affects prey more profoundly than previously believed and that its impact may not always be negative.     

Predator cat odors activate sexual arousal pathways in brains of Toxoplasma gondii infected rats

Cat odors induce rapid, innate and stereotyped defensive behaviors in rats at first exposure, a presumed response to the evolutionary pressures of predation. Bizarrely, rats infected with the brain parasite Toxoplasma gondii approach the cat odors they typically avoid. Since the protozoan Toxoplasma requires the cat to sexually reproduce, this change in host behavior is thought to be a remarkable example of a parasite manipulating a mammalian host for its own benefit. Toxoplasma does not influence host response to non-feline predator odor nor does it alter behavior on olfactory, social, fear or anxiety tests, arguing for specific manipulation in the processing of cat odor. We report that Toxoplasma infection alters neural activity in limbic brain areas necessary for innate defensive behavior in response to cat odor. Moreover, Toxoplasma increases activity in nearby limbic regions of sexual attraction when the rat is exposed to cat urine, compelling evidence that Toxoplasma overwhelms the innate fear response by causing, in its stead, a type of sexual attraction to the normally aversive cat odor.     

Sexual behaviour and its olfactory control in the desert woodrat (Neotoma lepida lepida)

In studies 1 and 2 nine pairs of desert woodrats (Neotoma lepida lepida) were observed for copulatory behaviour when the female was in a state of naturally occurring oestrus (study 1) and following ovariectomy and oestrogen and progesterone replacement (study 2). Males and females respond in a similar way under conditions of natural and hormone-induced oestrus. Males show a consummatory pattern involving multiple mounts and ejaculations, with ejaculations occurring after single intromissions. Females show the lordosis reflex accompanied by hop-and-dart and ear-wiggling responses. In addition, both sexes show appetitive precopulatory behaviours; the male emits an audible rasping vocalization as he trails and mounts the female, following a period of intense sniffing of the female's anogenital region. The female also frequently approaches and sniffs the male. In study 3, the role of female odours in the sexual behaviour of the male was examined in eight of the nine pairs used in studies 1 and 2. This was done by applying to the anogenital region of ovariectomized females a combination of urine and vaginal secretions taken from familiar and unfamiliar, and oestrogen-primed or non-oestrogen-primed females. The results show that odours from oestrogen-primed females are not sufficient to elicit male sexual behaviour, if the female is not sexually active. In study 4 the eight males were tested for their preferences for urine and vaginal secretion odours taken from females in different reproductive states and applied to cotton swabs. These males spent more time sniffing unfamiliar oestrous odours than unfamiliar non-oestrous odours and more time sniffing oestrous odours from a familiar female over those taken from an unfamiliar female.In study 5, 12 sexually active males were tested with oestrogen-primed females before and after either olfactory bulb removal or sham-surgery. Bulbectomized animals ceased copulating with females although females showed precopulatory approaches.Taken together, these studies suggest that normal sexual behaviour in the male woodrat requires that the female both possess the attractive odours (of oestrus) and that she engage in appetitive precopulatory behaviour.     

Functional activity of the rat olfactory bulb related to different durations of odor exposure

We have studied the effects of different odor exposure durationson the functional activity of the rat olfactory bulb by usingthe 2-deoxyglucose (2-DG) method. This technique brings outodor-specific patterns of labeling in the glomerular layer ofthe bulb. In a first set of experiments, rats designated ascontrols were submitted to two stimulation conditions with cyclohexanonefor 40 min following 2-DG injection: group C-5—alternatestimulation (5 min odor–5 mm pure air); group C-20—continuousstimulation (20 mm odor-20 min pure air). In a second set ofexperiments, rats designated as ‘adapted’ were exposedfor 3 h to cyclohexanone before the 2-DG injection; then, threeconditions of stimulation were tested with the same odor duringthe post-injection period; group Adap-5—alternate stimulation;group Adap-20—continuous stimulation; group Adap-0—stimulationwith pure air only. In the two control groups, the total numbersof labeled glomerular foci were similar; however, group C-20showed a significant increase in the number of ‘heavily’labeled foci. This result suggests that under conditions of20 min continuous odor exposure, receptor cells do not adaptrapidly and totally. Results from the adapted groups showedthat only the group Adap-20 presented significant changes infunctional bulbar activity. An important decrease in the totalnumber of labeled glomerular foci and an absence of ‘heavily’labeled foci were noted in all rats of this group; their patternsof 2-DG uptake were greatly reduced in both complexity and contrast.These results highlight the importance of olfactory receptoradaptation under conditions of long-duration odor exposure.Fast recovery of receptor sensitivity when the stimulation isinterrupted is also suggested.     

Mechanisms operating in the action of pheromones on the estrous cycle of the guinea-pig]

An increase in follicular growth was observed on the 6th and 8th days of the period of vaginal closure (PVC) in adult cyclic female guinea-pigs exposed to the odor of male urine from the beginning of this period. No statistically significant decrease in peripheral blood progesterone level occured on the same days of the PVC following exposure to the odor of male urine.     

Airborne manganese exposure differentially affects end points of oxidative stress in an age-and sex-dependent manner

Juvenile female and male (young) and 16-mo-old male (old) rats inhaled manganese in the form of manganese sulfate (MnSO₄) at 0, 0.01, 0.1, and 0.5 mg Mn/m³ or manganese phosphate at 0.1 mg Mn/m³ in exposures of 6h/d, 5d/wk for 13 wk. We assessed biochemical end points indicative of oxidative stress in five brain regions: cerebellum, hippocampus, hypothalamus, olfactory bulb, and striatum. Glutamine synthetase (GS) protein levels, metallothionein (MT) and GS mRNA levels, and total glutathione (GSH) levels were determined for all five regions. Although most brain regions in the three groups of animals were unaffected by manganese exposure in terms of GS protein levels, there was significantly increased protein (p<0.05) in the hippocampus and decreased protein in the hypothalamus of young male rats exposed to manganese phosphate as well as in the aged rats exposed to 0.1 mg/m³ MnSO₄. Conversely, GS protein was elevated in the olfactory bulb of females exposed to the high dose of MnSO₄. Statistically significant decreases (p<0.05) in MT and GS mRNA as a result, of manganese exposure were observed in the cerebellum, olfactory bulb, and hippocampus in the young male rats, in the hypothalamus in the young female rats, and in the hippocampus in the senescent males. Total GSH levels significantly (p<0.05) decreased in the olfactory bulb of manganese exposed young male rats and increased in the olfactory bulb of female rats exposed to manganese. Both the aged and young female rats had significantly decreased (p<0.05) GSH in the striatum resulting from manganese inhalation. The old male rats also had depleted GSH levels in the cerebellum and hypothalamus as a result, of the 0.1-mg/m³ manganese phosphate exposure. These results demonstrate that age and sex are variables that must be considered whenassessing the neurotoxicity of manganese.     

Spatially selective alteration of the mitral cell layer: a critical review of the literature

Evidence that chronic odor exposure alters mitral cell morphologyin young rats is reviewed. The functional origin of the mitralcell alteration is discussed in conjunction with the developmentof the olfactory bulb. It is concluded that odor exposure inyoung rats can alter the mitral cell layer in spatially selectivezones. Mitral cell alteration (primarily measured as penkaryaldiminution) appears to be a consequence of reduced stimulationand may be related to other activity-dependent ontogenetic effects.Chronic odor exposure provides an approach by which both spatialcoding of odor quality and functional development in the mammalianolfactory bulb can be examined.