Olfactory, partner preference and Fos expression in the vomeronasal projection pathway of sexually sluggish male rats

Sexually sluggish (S) male rats take a long time to ejaculate or sometimes they don't achieve ejaculation when tested on repeated occasions with receptive females. In order to further understand what factors might contribute to the inconsistent display of sexual behavior in these animals and determine if S and non-copulating males have a different neurobiological profile, the present study was design to characterize sex-related behaviors of S male rats. We tested their preference to physically interact between a sexually receptive female and a sexually active male. We also tested whether S males have a preference to investigate soiled bedding from females in estrous, anestrous or clean bedding. We also evaluated if the serum hormonal levels of testosterone (T) and estradiol (E) are different between copulating (C) and S male rats. Finally, we compared the neuronal activity of the vomeronasal projection pathway of C and S male rats exposed to estrous bedding. The results indicate that S male rats increased the percentage of animals displaying mounts and intromissions after repeated testing with receptive females but no increased was observed in the percentage of animals displaying ejaculations (a total of nine tests were performed). Both C and S males had a clear preference to interact with a sexually receptive female. Both groups showed a clear preference for bedding from estrous females as opposed to anestrous or clean bedding. However, the time investigating the estrous bedding was significantly lower for the S in comparison to C rats. No differences in the serum levels of T and E were found. The structures of the vomeronasal projection pathway were equally activated by estrous bedding in C and S male rats. The results indicate that S male rats do not have hormonal alterations or deficient processing of sexually relevant olfactory cues. Although S males showed a reduced preference for estrous bedding, no changes in preference for a receptive female were observed.     

Neuronal Fos activation in olfactory bulb and forebrain of male rats having erections in the presence of inaccessible estrous females.

Volatile odors from estrous female rats are necessary and sufficient to induce non-contact penile erections in male rats. It is not known whether these pheromones are detected by the accessory as opposed to the main olfactory system or whether they are processed by forebrain regions that receive olfactory inputs. Using nuclear Fos immunoreactivity as a marker of neuronal activation, we asked how the detection and processing of distal cues from inaccessible estrous females, which elicited non-contact penile erections, compared with the processing of sensory cues from soiled estrous bedding which did not elicit non-contact penile erections. In Experiment 1, groups of sexually experienced males were given one of five treatments. A control group was placed on clean bedding. A second group displayed non-contact penile erections when exposed to the smell, sight and sound of an estrous female restrained behind a permeable barrier. A third group was exposed to the same stimuli as the second (an estrous female) but failed to exhibit non-contact penile erections during the first hour of testing. A fourth group was placed on soiled estrous bedding, and a fifth group was allowed two ejaculations with an estrous female. All males were perfused with 4% paraformaldehyde 2 h after the onset of these respective treatments, and their brains were later processed for Fos immunoreactivity. Non-contact penile erections were observed in males that were exposed to distal cues from an estrous female but not in males exposed to soiled estrous bedding. Males that displayed non-contact penile erections or that were exposed to estrous bedding showed significantly more neuronal Fos immunoreactivity than clean-bedding controls in the nucleus accumbens core and shell, anterior and posterior medial amygdala, bed nucleus of the stria terminalis and the medial preoptic nucleus. Even greater neuronal Fos responses occurred in these regions in mated males. In Experiment 2 these same treatments were given to another cohort of sexually experienced males. Increased neuronal Fos immunoreactivity was observed in the granule and mitral cell layers of the accessory olfactory bulb of males that were either mated or exposed to estrous bedding, but not in males that displayed non-contact penile erections in response to distal cues from an estrous female. The volatile odors which presumably caused non-contact penile erections failed to stimulate significant neuronal Fos immunoreactivity in five main olfactory bulb sites examined. Even so, it seems likely that these pheromones are detected via the main olfactory system and are subsequently processed by the same projection circuit that responds to other pheromones present in estrous bedding that are incapable of eliciting non-contact penile erections.     

Olfactory environment and early mating behavior in the cyclic female rat

The objectives of this study were to examine the effects of olfactory cues from the male and of olfactory bulb removal on early mating behavior in sexually inexperienced diestrous female rats primed with estrogen. Four-day cyclic rats isolated from the male were given either 2.5 micrograms or 10 micrograms estradiol benzoate (EB) and presented to stimulated males in the late afternoon of diestrus 2 between 18:00 and 19:00 for a 10 min sexual behavioral session. Dose-dependent effects of estrogen were observed since 10 micrograms EB significantly increased the proportion of females displaying early mating as compared with those given 2.5 micrograms EB. Olfactory bulb removal prior to estrogen treatment caused a rise in the number of females which mated early with respect to the non bulbectomized controls. Exposing the females to bedding soiled with male urine on diestrus 2 at 10:00 did not affect early mating behavior. By contrast the olfactory stimuli became efficient when 2.5 micrograms EB treated females were given 10 micrograms progesterone (P) by the time of exposure to male urine. The results were discussed with respect to the role played by the olfactory system in the control of lordosis behavior throughout estrous cycle in female rats. P was concluded to be involved in the perception of the olfactory signals from the male which facilitate early mating behavior in diestrous female rats.     

Effects of neonatal handling on the behavior and prolactin stress response in male and female rats at various ages and estrous cycle phases of females

Neonatal handling induces behavioral and hormonal changes, characterized by reduced fear in novel environments, and lesser elevation and faster return to basal levels of plasma corticosterone, prolactin and adrenaline, in response to stressors in adulthood. The present study aimed to analyze the effects of neonatal handling from Days 1 to 10 postnatal on prolactin response to ether stress in male and female rats at three life periods: neonatal, peripubertal and adulthood. Moreover, adult females were tested in two different phases of the estrous cycle, i.e., diestrus and estrus. In another set of experiments, the behavior of peripubertal and adult males and females in estrus and diestrus was analyzed in the elevated plus maze test. Pups were either handled for 1 min (handled group) or left undisturbed (nonhandled group) during the first 10 days after delivery. In adults, in the handled females in diestrus, stress induced a lesser increase in plasma prolactin compared with nonhandled ones, as in males. However, in estrus, handled females showed no difference in the prolactin response to stress. In the elevated plus maze, handled females in diestrus, but not in estrus, showed higher locomotor activity compared with nonhandled ones. Peripubertal male and female rats handled during the neonatal period showed no difference in behavior in the elevated plus maze compared with nonhandled animals. Early-life stimulation can induce long-lasting behavioral and stress-related hormonal changes, but they are not stable throughout life and phases of the estrous cycle.     

Volatile female odors activate the accessory olfactory system of male mice without physical contact

We previously reported that male mice are more attracted to volatile odors from intact female mice than from ovariectomized female mice. In the present study, we investigated male attraction to volatile odors from soiled bedding collected from the cages of estrous or ovariectomized female mice. There was no difference in the total time spent sniffing volatile odors from estrous and ovariectomized female mice, suggesting that female mice emit volatile odors which are not excreted into bedding. To test this possibility, we investigated c-Fos expression in the mitral cell layer and granule cell layer of the accessory olfactory bulb 60 min after exposure of male mice to volatile odors without physical contact. Volatile odors from an estrous female mouse significantly increased the total number of c-Fos positive cells in each of the rostral and caudal granule cell layer, but not in the mitral cell layer. After exposure to volatile odors from estrous bedding, the total number of c-Fos positive cells did not increase. Volatile odors from a male mouse did not increase the total number of c-Fos positive cells. Volatile odors from an ovariectomized female mouse increased c-Fos expression only in the caudal granule cell layer. These results suggest that female mice emit specific volatile odors which are not excreted into bedding, and that the volatile odors activate the accessory olfactory system of male mice without physical contact. To characterize the female-specific volatile odors, we conducted habituation-dishabituation tests. Whereas sham-operated male mice discriminated between volatile odors of estrous and ovariectomized female mice, vomeronasal organ-removed male mice did not. These results suggest that male mice discriminated whether or not female mice were ovariectomized, by volatile odors via the accessory olfactory system, and that the female-specific volatile odors are involved in reproduction.     

Olfactory preference and Fos expression in the accessory olfactory system of male rats with bilateral lesions of the medial preoptic area/anterior hypothalamus

In the present study we evaluated if a medial preoptic area/anterior hypothalamus lesion affects the olfactory preference toward soiled bedding from receptive females in comparison to bedding from anestrous females or clean bedding. In the second part of the study we evaluated the accessory olfactory system response to estrous bedding with Fos immunoreactivity to determine if the preoptic lesions modify the processing of sexually relevant olfactory cues. Before medial preoptic area/anterior hypothalamus lesions, male rats spent more time investigating estrous bedding as opposed to anestrous or clean bedding. After the lesion, subjects showed no preference between estrous and anestrous bedding; that is, males spent the same amount of time investigating both types of bedding. These two odors were investigated more than clean bedding. Increments in Fos immunoreactivity neurons were seen in structures of the accessory olfactory system after exposure to soiled estrous bedding [granular layer of the accessory olfactory bulb, anterior-dorsal medial amygdala, posterior-dorsal medial amygdala, bed nucleus of the stria terminalis]. These results suggest that bilateral destruction of the medial preoptic area/anterior hypothalamus modify male olfactory preference in such a way that subjects spend the same time smelling and investigating bedding from estrous and anestrous females. This change in olfactory preference is not associated with alterations in the processing of sexually relevant olfactory cues by the accessory olfactory system.     

Urinary marking in male house mice: responses to novel environmental and social stimuli

The frequency of urinary marking by male house mice was investigated in two-chambered cages during exposure to either novel or familiar social or other environmental cues. In four experiments isolated males marked at high rates during their first exposure to the test cage (300–400 marks/hr), but showed significant habituation during subsequent testing (100–200 marks/hr on the fourth daily test). Marking frequency increased significantly when test-habituated males were placed in the presence of (in order of decreasing effectiveness): estrous females, ovariectomized females, castrate males, intact males, deermouse males, or male mouse urine. When all experiments were considered, however, there was little evidence that the various stimulus animals differentially affected marking rates. Male house mice also readily habituated to the presence of another male. Such male-habituated males then displayed a significant, but not dramatic, elevation of marking in the presence of an estrous female or a male deermouse, but they were totally unresponsive to the presence of a strange, test-habituated male. The results establish the importance of physical and/or biological novelty in eliciting urinary marking in male Mus.     

Serum corticosterone in fetal mice: sex differences, circadian changes, and effect of maternal stress.

The serum concentration of corticosterone was examined in control and stressed pregnant female mice (Mus domesticus) as well as male and female fetuses due to our interest in the behavioral effects of material stress on offspring in mice. Pregnant females were restrained under flood lights (2 sessions/day, 45 min/session) from Day 13-17 of pregnancy. On Day 17 of pregnancy a significant increase in maternal serum corticosterone was exhibited 1 h after the onset of a stress session, and serum corticosterone did not return to baseline until 16 h later. We also observed a significant increase in serum corticosterone in male fetuses during the first 4 h after maternal stress, while no significant change in serum concentration of corticosterone was observed in female fetuses throughout 24 h after maternal stress. Daily variation in serum concentration of corticosterone was also determined at 4-h intervals in pregnant mice and their fetuses from Day 16-18 of pregnancy. Pregnant females maintained on a 12 L:12 D cycle exhibited peak serum corticosterone concentrations at and just before the onset of the darkness. Daily fluctuations in serum concentrations of corticosterone in male and female fetuses reflected the pattern observed in the mothers. A sex difference in serum corticosterone in fetuses was observed at some, but not all times of the day, with the difference being greatest during the dark phase of the mother's light:dark cycle.     

Sexual and olfactory preference in noncopulating male rats

In some species including rats, mice, gerbils, and rams, apparently normal males fail to copulate when repeatedly tested with receptive females. These animals are called "noncopulators (NC)," and the cause of this behavioral deficit is unknown. It has been shown that NC rats do not have hormonal alterations or deficits in the mechanisms that control penile function. The present study was designed to examine (Experiment 1) whether NC male rats prefer receptive females to sexually active males. In addition, the olfactory preference for bedding soiled from estrous or for anestrous bedding was investigated. These tests were performed in NC and copulating (C) male rats when the subjects were intact, gonadectomized (GDX), or GDX and treated with high doses of testosterone propionate (TP). Our results demonstrate that NC rats do not display sexual behavior even after high TP treatment. While C male rats have a clear preference for receptive females as opposed to a sexually active male, NC rats do not. In all hormonal conditions, the preference shown by NC rats for estrous bedding was significantly reduced in comparison to that seen in C rats. TP treatment in NC rats did not modify either partner or odor preference. In Experiment 2, we evaluated if NC rats are feminized and if it could be easier to induce feminine-like behavior by hormone treatment with estradiol benzoate (EB) or with EB plus progesterone (P) (EB+P). Odor preference for estrous or male bedding under these hormonal conditions was also compared. No differences between NC and C rats were found in feminine sexual behavior. In the olfactory test, we found that NC rats prefer odors from receptive females as opposed to male odors, but this preference is reduced compared to that of C rats. Males treated with EB or EB+P show no preference for female odors. These results demonstrate that treatment with EB or EB+P does not increase feminine sexual behavior in NC rats.     

Attraction to male pheromones and sexual behaviour show different regulatory mechanisms in female mice

In rodents, female sexual behaviour is under hormonal control. The attraction females show for male-derived nonvolatile chemicals (pheromones) can be regarded as the first step of this behaviour, but it is unknown whether this attraction is also modulated by sexual steroids. To test this possibility, ovariectomized adult female mice with no experience of chemical signals from adult males were randomly assigned to four groups that received oil (control), progesterone, estradiol (E) or estradiol+progesterone (E+P) injections, respectively. Females were then tested for their attraction to male-soiled bedding and, subsequently, for their proceptive behaviour when confronted to adult males. Females showed attraction to male-soiled bedding irrespective of the hormonal treatment, whereas only those females treated with E or E+P showed proceptive behaviour. Therefore, in contrast to proceptive and copulatory behaviour, the female attraction to male pheromones is independent of sexual steroids, thus indicating that those parts of the vomeronasal system involved in this attraction do not respond to steroids. In summary, sexual behaviour in female mice can be seen as a two-step process. First, females are attracted by male pheromones, a process which is independent of their hormonal status. After encountering the males, females show proceptive behaviour only in estrous, when fertilization is more likely. The attraction exerted by male sexual pheromones promotes female autostimulation that might ensure anticipatory endocrine changes leading to ovulation by the time of sexual intercourse.     

Activation of aggression in female rats by normal males and by castrated males with testosterone implants

Female hooded rats were continuously housed with an intact male, a castrated male with subcutaneous testosterone implants, or two other females. At weekly intervals over a 10-week period, the cagemate(s) and pups were removed and aggression by the female toward an unfamiliar female intruder was observed over a 15-min period. On the 11th week each female was subjected to this intruder test in an unfamiliar cage. On the 12th week, a final test was conducted in each female's living cage with a male rather than a female as the intruder. The aggressive behaviors recorded were attacks, bites, on-top, and piloerection. Females housed with normal males displayed a significant increase in aggression prior to parturition. Their aggressiveness persisted through the 10th test with peaks at parturition and the start of lactation. Females housed with castrated males also displayed significant increases in aggression but without the peaks associated with parturition and lactation. Their aggressiveness also persisted throughout the test period. Females housed with other females showed a small increase in aggression over weeks. All groups showed virtually no aggression in the unfamiliar cage. All females displayed some aggression toward a male intruder but the level of aggression was highest in maternal females. The results demonstrate that aggression qualitatively similar to that displayed following parturition and during lactation can be elicited in nulliparous females.     

Social stress in mice: gender differences and effects of estrous cycle and social dominance.

A large discrepancy in the possibility of inducing social stress in the two genders exists. Since generalizations of findings from one sex to the other appear not to be valid, reliable models of social stress in females are needed. We examined the effects of social context in the housing environment, as a possible source of stress, on exploration and anxiety in male and female mice, taking into account the estrous phase for females and the social status for males as additional variables. Mice housed individually or with siblings were tested in a free-exploratory paradigm of anxiety (where test animals have a choice to stay in their home cage or to explore an open field, OF). Individually housed females did not leave their home cage for long periods, explored less the unfamiliar area and displayed higher risk assessment, a behavioral profile suggestive of lower propensity for exploration and higher level of anxiety compared with group-housed females. Individually housed males tended to show an opposite profile. Proestrus mice were less sensitive to the decrease of exploratory propensity induced by individually housing compared to estrus and diestrus mice. Social dominants and social subordinates in sibling groups did not differ in their exploratory responses to the OF. Different housing procedures, as means to provide different social environment, may differentially induce mild social stress in male and female mice.     

Coexpression of sexual behavior and maternal aggression: the ambivalence of sexually active mother rats toward male intruders

The sexually active female rat solicits the male to approach for copulation, while the maternal dam displays aggression to expel him from the nest, suggesting that both behaviors are mutually exclusive. However, the rat has a postpartum estrus during which she is sexual and maternally motivated. Can she perceive the male as attractive and aversive, soliciting and attacking him at the same time? This study shows that postpartum estrous females exhibit a merge of sexual and maternal aggressive responses toward male intruders in the home cage. The concurrent expression of these behaviors did not affect their intensities, although the stimulation of maternal behavior increased maternal aggression without modifying sexual solicitation. These results indicate that the postpartum estrous rat can optimally express two opposite and independently regulated motivations, and that the male can be perceived as an ambivalent stimulus.     

Characteristics of the Behavior and Stress-Reactivity of the Hypophyseal-Adrenal System in Prenatally Stressed Rats

The effects of daily 1-h immobilization of female rats from days 15 to 18 of pregnancy on the levels of anxiety, orientational-investigative activity in an open field test, and the dynamics of the stress response of the hypophyseal-adrenal system were studied in the male and female adult offspring of these rats. Maternal stress was found to induce significant reductions in the level of orientational-investigative activity of females in the stage of diestrus, and to increase anxiety as measured in an elevated cross maze. Prenatally stressed males, conversely, had decreased levels of anxiety, and behavior in the open field test was virtually unaltered. As a result, prenatally stressed rats showed smoothing out of the intergender differences in these forms of behavior, seen in control animals in normal conditions. Prenatal stress had a significant influence on the dynamics of the stress response of the hypophyseal-adrenal system in males and females; males showed impairment of the feedback control of this system in conditions of stress activation, while females showed significant increases in the maximum level of adrenal corticosterone secretion into the blood in response to immobilization lasting 20 min. These data provide evidence that maternal stress has significant influences on behavior and the stress response in both male and female rats.     

Changes in male copulatory behavior after sexual exciting stimuli: effects of medial amygdala lesions.

A paradigm was developed to investigate how precoital sexual arousal affects parameters of sexual behavior in male rats. Estrous females in a wire mesh cage were used to induce sexual arousal before the sexual interaction test. In control procedures, males were presented in a wire mesh cage or else there was no stimuli at all. The results indicate that ejaculation latency is consistently reduced after preexposure to a female, but not after preexposure to a male, showing that the effect is specific for precoital sexual arousal. Other parameters were affected by precoital sexual arousal in some, but not in all experiments. Reductions in intromission latency moreover, were observed after both preexposure to a male and preexposure to a female, indicating that general social excitement affects this parameter. Preexposure to females for 10 minutes or 3 hours produced similar results. It was subsequently found that medial amygdala-lesioned (AME) animals differed from sham-lesioned (SHAM) controls with respect to their reaction to precoital sexual arousal. The results show that AME-lesioned animals, in contrast to SHAM-animals, do not show reduced ejaculation latencies after preexposure to an estrous female. The results are in line with the idea that AME-lesioned animals are deficient in the assimilation of information on sexual exciting stimuli.     

Male-induced estrus synchronization in the female Siberian hamster (Phodopus sungorus sungorus)

Olfactory cues play an integral role in the organization of events that mediate reproductive success. In a variety of species, priming pheromones, in particular, are important for ensuring reproductive fitness. To date, very little research has focused on how male-emitted priming pheromones, such as those that regulate the onset of puberty and estrus synchronization in females, affect the reproductive physiology of the female Siberian hamster (Phodopus sungorus sungorus). This lack of research may be due to the physiology of the Phodopus genus; vaginal cytology cannot be used as a reliable indicator of estrus or ovulation. Using a jugular cannulation technique to determine estrous stage by blood analysis of prolactin and luteinizing hormone, we sought to determine if male priming pheromones affect estrous cyclicity in the female Siberian hamster and, if so, whether the production of these priming pheromones is androgen dependent. Our results showed that females exposed to bedding from mature, intact males showed a significantly higher incidence of proestrus 3 days later than did females exposed to the bedding of mature, gonadectomized males. Therefore, we found that not only do male Siberian hamsters emit chemical signals that induce estrus synchronization, but also that this ability is likely to be androgen dependent.     

Reproductive status influences cell proliferation and cell survival in the dentate gyrus of adult female meadow voles: a possible regulatory role for estradiol

Galea and McEwen [Galea and McEwan (1999) Neuroscience 89, 955-964] found that cell proliferation was suppressed in female meadow voles trapped during the breeding season relative to females trapped during the non-breeding season. We investigated the effect of reproductive status and estradiol level on cell proliferation and cell survival in adult laboratory-reared female meadow voles to control for the variables of age, experience and pregnancy that could confound the results derived from a wild sample. Voles were housed in either a long- or short-photoperiod to simulate season and a male or female cage partner was introduced to influence reproductive status. Because females are reflex ovulators, exposure to a male rapidly induces behavioural estrous and high levels of estradiol. Forty-eight hours after introducing a cage partner, we injected either bromodeoxyuridine or [3H]thymidine to mark cell synthesis and then examined labelled cells 2h (cell proliferation) or five weeks (cell survival) later, respectively. To determine whether estradiol mimicked the effect of reproductive status, groups of reproductively inactive females were given a single injection of estradiol benzoate (10 microg) either four or 48h prior to bromodeoxyuridine labelling. The density of proliferating cells in the granule cell layer and the hilus was elevated in reproductively inactive females compared to reproductively active females and was correlated negatively with serum estradiol level. Exposure to estradiol benzoate initially increased cell proliferation (within 4h) but subsequently suppressed cell proliferation (within 48h). In addition, the density of surviving cells was greater in reproductively inactive females relative to reproductively active females but reproductively active females had a greater rate of cell survival than did reproductively inactive females. Reproductive status did not influence the number of pyknotic cells in the dentate gyrus (at either 2h or five weeks).We conclude that reproductive status regulates cell proliferation in adult female meadow voles, possibly via an estradiol-regulated mechanism. The results from the present study showed that reproductively active female meadow voles have suppressed rates of cell proliferation in the dentate gyrus relative compared with reproductively inactive female meadow voles. Administering estradiol initially (within 4h) elevates the cell proliferation within the dentate gyrus of adult females but subsequently (within 48h) suppresses cell proliferation. However, more new cells survived in females with high endogenous levels of estradiol (reproductively active females). In conclusion, reproductive status regulates the level of cell proliferation and survival through a complex estradiol regulated mechanism(s).     

Sexual maturation of female house mice: social inhibition

Young female laboratory mice reared in the presence of an adult male mouse or male bedding containing a pheromone reach sexual maturity earlier than control females. A pheromone produced by grouped female mice leads to long, irregular estrous cycles or anestrus. The present study demonstrated that a female-produced pheromone delays sexual maturation when females are grouped and that free social interaction and tactile contact among the grouped females are necessary for the production of this inhibitory pheromone. Both young and adult females caged in groups produced the inhibitory pheromone. This study provided additional support for the hypothesis that in female mice morphological and sexual development are under separate control mechanisms. Two pheromones may be active in reproductive processes of female mice: one exhibiting inhibitory effects and the other acceleratory effects.     

Copulation-illness associations in male rats: lithium chloride dose and delay manipulations

Male rats that receive an injection of lithium chloride (LiCl) after each pairing with an estrous female gradually decrease their copulatory behaviors. In the present experiments we demonstrated that a 6.0 mEq/kg (0.3 M, 20 ml/kg) dose of LiCl injected after trials spaced twice weekly at 3-4-day intervals induced more rapid acquisition of copulation-illness associations than a 3.0 mEq/kg (0.15 M, 20 ml/kg) dose. Contingent 0.3 M saline or noncontingent 0.3 M LiCl injections did not affect copulatory behaviors. The location of the male rat (home cage or test chamber) during a portion of the aversive state induced by LiCl did not influence rate of acquisition. Copulation-illness associations, once established, were retained over a 60-day interval. Comparable decrements in copulatory behaviors were evident when LiCl was injected at 1-, 5-, or 15-min intervals after pairings with estrous females when the males were detained in the test chambers during the delay intervals; decrements were not observed when the interval was increased to 30 or 60 min. An electric shock analog to the aversive state induced by LiCl did not induce decrements in copulatory behaviors. It was suggested that odor-illness associations may, in part, account for the decrements in copulatory behaviors in this paradigm.     

Hippocampal lesions are without effect on olfactory memory formation in the context of pregnancy block

Female mice which have mated and are subsequently exposed to male urine odours (pheromones), which differ from those of the male that mated (stud male), undergo hormonal changes resulting in a block to pregnancy. Since the stud male's odour blocks the pregnancy of females other than those he mated, this would suggest that a memory specific for this male's odour is established at the time of mating. Hippocampal lesions, when made prior to mating, did not disrupt memory formation to the odour of the stud male. This male's odour did not block pregnancy, while changing odour to that of the strange male did block pregnancy. In order to establish the functional effectiveness of these hippocampal lesions, olfactory discriminations were examined in a modified T-maze. The odour discrimination was urine-soiled bedding taken from a cage of different strain males versus same strain bedding. After 35 trials, neither control nor lesioned females were performing above chance level (50% correct responses), indicating the difficulty in cognitively learning this discrimination. These female mice were therefore trained to distinguish a novel odour (butyl acetate) from familiar strain urine-soiled bedding. Hippocampal-lesioned females were slower to acquire this discrimination and did not perform above chance level after 35 trials, in contrast to the 70% success of sham-lesioned females. Hence, hippocampal lesions, while producing a deficit in olfactory learning in the maze test, are without effect on the formation or retrieval of the specific olfactory memory formed for the stud male at mating, as revealed from pregnancy block tests.