Convergence of main and accessory olfactory pathways onto single neurons in the hamster amygdala

Summary Chemoreceptor pathways from the vomeronasal organ (VNO), and main olfactory system are known to be separate as they pass into the brain, at least until the level of the amygdala. In the amygdala, vomeronasal pathways project to the posteromedial cortical nucleus (PMCN), and medial nucleus (MN). The main olfactory pathways have terminations in the posterolateral cortical nucleus (PLCN), and anterior cortical nucleus (ACN), both of which project to the PMCN and MN. The anatomy thus suggests that the PMCN and MN are sites for convergence of input from the main and accessory olfactory pathways. We have recorded single units in the amygdala and found that electrical stimulation of either the main olfactory bulbs or the VNO could drive some of the same units in the PMCN. Units were also found that were driven by one system but not the other, and units in which activity driven by one system was suppressed by stimulation of the other system.     

Social contact elicits immediate-early gene expression in dopaminergic cells of the male prairie vole extended olfactory amygdala

Male prairie voles (Microtus ochrogaster) are a valuable model in which to study the neurobiology of sociality because, unlike most mammals, they pair bond after mating and display paternal behaviors. Research on the regulation of these social behaviors has highlighted dopamine (DA) neurotransmission in both pair bonding and parenting. We recently described large numbers of dopaminergic cells in the male prairie vole principal nucleus of the bed nucleus of the stria terminalis (pBST) and posterodorsal medial amygdala (MeApd), but such cells were very few in number or absent in the non-monogamous species we examined, including meadow voles. This suggests that DA cells in these sites may be important for sociosexual behaviors in male prairie voles. To gain some insight into the function of these DAergic neurons in male prairie voles, we examined expression of the immediate-early genes (IEGs) Fos and Egr-1 in tyrosine hydroxylase (TH)–immunoreactive (TH-ir) cells of the pBST and MeApd after males interacted or not with one of several social stimuli. We found that IEGs were constitutively expressed in some TH-ir neurons under any social condition, but that IEG expression in these cells decreased after a 3.5-h social isolation. Thirty-minute mating bouts (but not 6- or 24-h bouts) that included ejaculation elicited greater IEG expression in TH-ir cells than did non-ejaculatory mating, interactions with a familiar female sibling, or interactions with pups. Furthermore, Fos expression in TH-ir cells was positively correlated with the display of copulatory, but not parental, behaviors. These effects of mating were not found in other DA-rich sites of the forebrain (including the anteroventral periventricular preoptic area, periventricular anterior hypothalamus, zona incerta, and arcuate nucleus). Thus, activity in DAergic cells of the male prairie vole pBST and MeApd is influenced by their social environment, and may be particularly involved in mating and its consequences, including pair bonding.     

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.     

The role of the medial preoptic area in appetitive and consummatory reproductive behaviors depends on sexual experience and odor volatility in male Syrian hamsters

In Syrian hamsters (Mesocricetus auratus), the expression of reproductive behavior requires the perception and discrimination of sexual odors. The behavioral response to these odors is mediated by a network of ventral forebrain nuclei, including the medial preoptic area (MPOA). The role of MPOA in male copulatory behavior has been well-studied, but less is known about the role of MPOA in appetitive aspects of male reproductive behavior. Furthermore, many previous studies that examined the role of MPOA in reproductive behavior have used large lesions that damaged other nuclei near MPOA or fibers of passage within MPOA, making it difficult to attribute post-lesion deficits in reproductive behavior to MPOA specifically. Thus, the current study used discrete, excitotoxic lesions of MPOA to test the role of this nucleus in opposite-sex odor preference and copulatory behavior in both sexually-naïve and sexually-experienced males. Lesions of MPOA eliminated preference for volatile, opposite-sex odors in sexually-naïve, but not sexually-experienced, males. When males were allowed to contact the sexual odors, however, preference for female odors remained intact. Surprisingly, lesions of MPOA caused severe copulatory deficits only in sexually-naïve males, suggesting previous reports of copulatory deficits following MPOA lesions in sexually-experienced males were not due to damage to MPOA itself. Together, these results demonstrate that the role of MPOA in appetitive and consummatory aspects of reproductive behavior varies with the volatility of the sexual odors and the sexual experience of the male.     

Intra-hypothalamic melatonin blocks photoperiodic responsiveness in the male Syrian hamster

Exposure of male Syrian hamsters to a short daylength of 8L:16D leads to gonadal regression. This effect of photoperiod was prevented by pinealectomy or chronic exposure of the brain to exogenous melatonin delivered from in-dwelling cannulae. However, the effect of melatonin was dependent on the neural site of application. Melatonin delivered into the mid-brain, lateral hypothalamus or amygdala was ineffective. In contrast, bilateral administration of melatonin to the medial or amygdala was ineffective. In contrast, bilateral administration of melatonin to the medial hypothalamus prevented testicular regression and maintained high circulating levels of luteinizing hormone and prolactin. These findings suggest that the medial hypothalamus contains target sites for melatonin involved in pineal-mediated photoperiodic responses.     

The effects of medial preoptic area and amygdala lesions on maternal behavior in the juvenile rat

The present study was designed to determine whether the medial preoptic area (MPOA) and the amygdala (AMYG) are involved in the expression of "maternal" behavior in juvenile rats as they are in the adult. Juveniles show many behaviors that are similar to the maternal behaviors shown by the postpartum female rat. Whether these behaviors are social in function, as opposed to parental, and hence mediated by different mechanisms from those regulating adult maternal behavior is not known. To test the roles of the MPOA and AMYG in mediating these behaviors, 21-day-old female juvenile rats received MPOA, AMYG, or SHAM (MPOA/AMYG) lesions and were tested at 22 days of age for maternal and other responses to pups. Major findings demonstrate that MPOA lesions disrupt components of maternal behavior, including retrieving and nest building, while AMYG lesions facilitate these behaviors. These findings indicate striking similarities between the juvenile and rat brain for parental responding.     

The posteromedial cortical amygdala regulates copulatory behavior, but not sexual odor preference, in the male Syrian hamster (Mesocricetus auratus)

In rodent species, the expression of reproductive behavior relies heavily on the perception of social odors, as well as the presence of circulating steroid hormones. In the Syrian hamster, chemosensory and hormonal cues are processed within an interconnected network of ventral forebrain nuclei that regulates many aspects of social behavior. Within this network, the posteromedial cortical amygdala (PMCo) receives direct projections from the accessory olfactory bulbs and contains a dense population of steroid receptor-containing neurons. Consequently, the PMCo may be important for generating odor-guided aspects of reproductive behavior, yet little is known regarding the role of this nucleus in regulating these behaviors. Thus, the present study tested male hamsters with site-specific electrolytic lesions of the PMCo for their (a) sexual odor preference in a Y-maze apparatus, (b) sexual odor discrimination in a habituation-dishabituation task, and (c) copulatory behavior when paired with a sexually receptive female. PMCo-lesioned males preferred to investigate female odors over male odors and were able to discriminate between these odor sources. However, PMCo lesions were associated with several alterations in the male copulatory pattern. First, PMCo-lesioned males displayed increased investigation of the female's non-anogenital region, suggesting that the PMCo may be involved in directing appropriate chemosensory investigation during mating. Second, PMCo lesions altered the temporal pattern of the mating sequence, as PMCo-lesioned males took longer than Sham-lesioned males to reach sexual satiety, as indicated by the delayed expression of long intromissions. This delayed onset of satiety was associated with an increased number of ejaculations compared with Sham-lesioned males. Importantly, these data provide the first direct evidence for a functional role of the PMCo in regulating male reproductive behavior.     

Nitric oxide mediates glutamate-evoked dopamine release in the medial preoptic area

Dopamine (DA) release in the medial preoptic area (MPOA) of the hypothalamus is an important facilitator of male sexual behavior. The presence of a receptive female increases extracellular DA in the MPOA, which increases further during copulation. However, the neurochemical events that mediate the increase of DA in the MPOA are not fully understood. Here we report that glutamate, reverse-dialyzed into the MPOA, increased extracellular DA, which returned to baseline after the glutamate was removed. This increase was prevented by co-administration of the nitric oxide synthase inhibitor NG-nitro-l-arginine methyl ester (L-NAME), but not by the inactive isomer, Nw-nitro-d-arginine methyl ester (D-NAME). In contrast, extracellular concentrations of the major metabolites of DA were decreased by glutamate, suggesting that the DA transporter was inhibited. These decreases were also inhibited by L-NAME, but not D-NAME. These results indicate that glutamate enhances extracellular DA in the MPOA, at least in part, via nitric oxide activity. Therefore, glutamatergic stimulation of nitric oxide synthase may generate the female-induced increase in extracellular DA in the MPOA, which is important for the expression of male sexual behavior.     

Capillary-to-fiber ratio of hind limb muscles in the male Syrian golden hamster

The hamster has been the accepted model of emphysema since the 1970s, demonstrating disease-related effects on respiratory skeletal muscle. However, there is scant information available about the model's ability to replicate the peripheral skeletal muscle changes seen in human disease, such as alterations in capillarity. The present study described the capillary-to-fiber ratio (C/F) of normal hamster plantaris, gastrocnemius, and soleus muscles in eight animals. C/F was 1.72 +/- 0.38 for plantaris, 1.95 +/- 0.40 for gastrocnemius, and 2.22 +/- 0.43 for soleus. C/F of soleus was significantly greater (P < 0.01) than plantaris. The C/F of hamster hindlimb muscles varies from those seen in rat species, and having baseline data on hamsters makes it possible to determine the effects of emphysema on C/F in this model.     

Increased expression of c-fos in the medial preoptic area after mating in male rats: role of afferent inputs from the medial amygdala and midbrain central tegmental field.

Immunocytochemical methods were used to localize the protein product of the immediate-early gene, c-fos, in male rats after exposure to, or direct physical interaction with, oestrous females. Increasing amounts of physical contact with a female, with resultant olfactory-vomeronasal and/or genital-somatosensory inputs, caused corresponding increments in c-fos expression in the medial preoptic area, the caudal part of the bed nucleus of the stria terminalis, the medial amygdala, and the midbrain central tegmental field. Males bearing unilateral electrothermal lesions of the olfactory peduncle showed a significant reduction in c-fos expression in the ipsilateral medial amygdala, but not in other structures, provided their coital interaction with oestrous females was restricted to mount-thrust and occasional intromissive patterns due to repeated application of lidocaine anaesthetic to the penis. No such lateralization of c-fos expression occurred in other males with unilateral olfactory lesions which were allowed to intromit and ejaculate with a female. These results suggest that olfactory inputs, possibly of vomeronasal origin, contribute to the activation of c-fos in the medial amygdala. However, lesion-induced deficits in this type of afferent input to the nervous system appear to be readily compensated for by the genital somatosensory input derived from repeated intromissions. Unilateral excitotoxic lesions of the medial preoptic area, made by infusing quinolinic acid, failed to reduce c-fos expression in the ipsilateral or contralateral medial amygdala or central tegmental field following ejaculation. By contrast, combined, unilateral excitotoxic lesions of the medial amygdala and the central tegmental field significantly reduced c-fos expression in the ipsilateral bed nucleus of the stria terminalis and medial preoptic area after mating; no such asymmetry in c-fos expression occurred when lesions were restricted to either the medial amygdala or central tegmental field. This suggests that afferent inputs from the central tegmental field (probably of genital-somatosensory origin) and from the medial amygdala (probably of olfactory-vomeronasal origin) interact to promote cellular activity, and the resultant induction of c-fos, in the ipsilateral bed nucleus of the stria terminalis and medial preoptic area. The monitoring of neuronal c-fos expression provides an effective means of studying the role of sensory factors in governing the activity of integrated neural structures which control the expression of a complex social behaviour.     

Polymodal activation of the endocannabinoid system in the extended amygdala

The reason why neurons synthesize more than one endocannabinoid (eCB) and how this is involved in the regulation of synaptic plasticity in a single neuron is not known. We found that 2-arachidonoylglycerol (2-AG) and anandamide mediate different forms of plasticity in the extended amygdala of rats. Dendritic L-type Ca(2+) channels and the subsequent release of 2-AG acting on presynaptic CB1 receptors triggered retrograde short-term depression. Long-term depression was mediated by postsynaptic mGluR5-dependent release of anandamide acting on postsynaptic TRPV1 receptors. In contrast, 2-AG/CB1R-mediated retrograde signaling mediated both forms of plasticity in the striatum. These data illustrate how the eCB system can function as a polymodal signal integrator to allow the diversification of synaptic plasticity in a single neuron.     

Procaine hydrochloride olfactory block eliminates mounting in the male golden hamster

Intranasal profusion with procaine hydrochloride eliminated mounting and subsequent mating behavior in both sexually experienced and sexually inexperienced male golden hamsters (Mesocricetus auratus). Mounting behavior of control groups receiving intranasal application of tap water alone or in combination with intraperitoneally injected procaine hydrochloride was not markedly effected. Bouts of vaginal licking and sniffing were depressed by procaine hydrochloride, but not by tap water. These results indicate the elimination of mounting following olfactory bulbectomy in this species need not be explained on the basis of neurological degeneration to more central brain centers.     

Dopamine, the medial preoptic area, and male sexual behavior

The medial preoptic area (MPOA), at the rostral end of the hypothalamus, is important for the regulation of male sexual behavior. Results showing that male sexual behavior is impaired following MPOA lesions and enhanced with MPOA stimulation support this conclusion. The neurotransmitter dopamine (DA) facilitates male sexual behavior in all studied species, including rodents and humans. Here, we review data indicating that the MPOA is one site where DA may act to regulate male sexual behavior. DA agonists microinjected into the MPOA facilitate sexual behavior, whereas DA antagonists impair copulation, genital reflexes, and sexual motivation. Moreover, microdialysis experiments showed increased release of DA in the MPOA as a result of precopulatory exposure to an estrous female and during copulation. DA may remove tonic inhibition in the MPOA, thereby enhancing sensorimotor integration, and also coordinate autonomic influences on genital reflexes. In addition to sensory stimulation, other factors influence the release of DA in the MPOA, including testosterone, nitric oxide, and glutamate. Here we summarize and interpret these data.     

Quantitative autoradiographic analysis of estradiol retention by cells in the preoptic area,hypothalamus and amygdala

Summary These experiments were done to compare quantitatively, on a cell-by-cell basis, estradiol retention by cells in the medial preoptic area, arcuate nucleus, ventrolateral subdivision of the ventromedial nucleus, and the caudal half of the medial nucleus of the amygdala. The steroid autoradiograms were prepared from 2 sections of brains from ovariectomized, adrenalectomized adult female rats that had been infused intravenously with [³H] estradiol (E₂) in a regimen which kept circulating hormone concentration at or above proestrus levels for 3–4 h. Even in these brain regions, containing the most dense collections of E₂-concentrating cells, a maximum of only 27–61% of the cells concentrated E₂. Therefore, in these regions only a particular subset of the cells retain hormone; other cells in the region do not retain hormone. Frequency distribution histograms of the number of grains per cell versus the number of cells in each region showed a wide range in the amount of E₂ retained per cell, and no modes among E₂retaining cells. The data followed a distribution markedly different from that predicted by a simple Poisson distribution, confirming that E₂-retention does not result from a random, passive process such as diffusion. The overall quantitative characteristics of the frequency distribution histograms were similar across the four brain areas. Therefore, we propose that the different E₂-sensitive functions of these brain areas must depend on differences in the neural connectivity or differences in hormone regulated peptide content of the areas.     

Mating behavior induces selective expression of Fos protein within the chemosensory pathways of the male Syrian hamster brain.

The effect of mating behavior on the expression of Fos protein was analyzed within the chemosensory pathways of the male Syrian hamster brain. Following a single mating test, the number of Fos-immunoreactive (Fos-ir) neurons increased within the amygdala, bed nucleus of the stria terminalis and medial preoptic area. The mating-induced pattern of Fos expression within these brain regions shows a strong correlation with the sites of lesions that eliminate or alter mating behavior. In addition, Fos expression was increased within the paraventricular nucleus of the hypothalamus. These results provide the first demonstration of a dynamic and selective pattern of neuronal activity within specific nuclei known to be essential for mating behavior in the male Syrian hamster.     

Social novelty increases tyrosine hydroxylase immunoreactivity in the extended olfactory amygdala of female prairie voles

The monogamous social behaviors of prairie voles (Microtus ochrogaster) require olfactory inputs, which are processed by the posterodorsal medial amygdala (MeApd) and principal bed nucleus of the stria terminalis (pBST). The male prairie vole MeApd and pBST contain hundreds of cells densely immunoreactive for tyrosine hydroxylase (TH-ir). Female prairie voles have relatively few of these cells, but we previously found that the number of these TH-ir cells is greatly increased in females by exogenous estradiol. We here hypothesized that the number of TH-ir cells in their MeApd and pBST would also increase during the natural hormone surges associated with females' induced estrus. We found that the number of TH-ir cells in both sites did significantly increase after females cohabitated for two days with an unfamiliar male. However, this increase did not require the presence of ovaries and even tended to occur in the pBST of females cohabitating for two days with unfamiliar females. We then determined if the greater number of TH-ir cells after heterosexual pairing was transient by examining two groups of long-term pairbonded females (primiparous and multiparous), and found these females also had significantly more TH-ir cells in the pBST and/or MeApd compared to unmated controls. Thus, social novelty arising from cohabitation with unfamiliar conspecifics produces a reoccurring increase in the number of TH-ir cells in the female prairie vole extended olfactory amygdala. Ovarian hormones are not necessarily required. This increase in catecholaminergic cells may facilitate acquisition and retention of olfactory memories necessary for social recognition in this species.     

Mating activates NMDA receptors in the medial preoptic area of male rats

Studies have emphasized the role of the medial preoptic area (MPOA) as an important site for the regulation of male sexual behavior. Indeed, ablations of the MPOA impair sexual behavior, whereas stimulation of the MPOA enhances behavior. Furthermore, neural activity in the MPOA increases with mating. The current study tested the hypothesis that activation of N-methyl-D-aspartate (NMDA) receptors occurs in MPOA neurons and is essential for the expression of male sexual behavior in rats. Results indicate that nearly all MPOA neurons that expressed Fos following mating also contained the NR1 subunit of NMDA receptors. Furthermore, mating increased phosphorylation, thus activation, of NR1 in the MPOA. Additionally, blocking NMDA receptors significantly decreased mating-induced Fos expression and mating-induced phosphorylation of NMDA receptors and impaired male sexual behavior. These results provide evidence that mating activates NMDA receptors in the MPOA and that this activation is important for the expression of male sexual behavior.     

The effects of castration, testosterone replacement and photoperiod upon hypothalamic beta-endorphin levels in the male Syrian hamster

Syrian hamsters kept in long day-lengths have active gonads and high circulating levels of gonadal steroids. Under the influence of the pineal gland, animals exposed to short photoperiods undergo testicular regression, have low circulating levels of testosterone and gonadotrophins and elevated levels of beta-endorphin within the hypothalamus. This paper describes the interaction between testosterone and photoperiod in the regulation of beta-endorphin levels in three regions of the hypothalamus. Hypothalamic beta-endorphin levels were measured by a combination of high-performance liquid chromatography and radioimmunoassay techniques that allows separation of the beta-endorphin (1-31) peptide from its metabolites and precursors. All of the beta-endorphin-like immunoreactivity in the hypothalamus of the male hamster, in both photoinhibited and photostimulated conditions, was found to represent the 31-amino-acid peptide. In photostimulated hamsters, chronic castration was associated with a significant increase of beta-endorphin levels in the anterior hypothalamus and mediobasal hypothalamus, which was reversed by treatment with exogenous testosterone. Castration prevented the ability of naloxone, an opiate receptor antagonist, to release luteinizing hormone, and this effect was also reversed by exogenous steroid. In photoinhibited hamsters, however, castration had no effect upon beta-endorphin levels in the preoptic area or mediobasal hypothalamus, and there was only a small increment in the anterior hypothalamus. Significantly, beta-endorphin levels in all areas of the hypothalamus of photoinhibited castrates were not decreased by testosterone treatment. In addition, administration of exogenous testosterone did not restore sensitivity to naloxone in these animals.(ABSTRACT TRUNCATED AT 250 WORDS)