Projection Sites of Medial Preoptic Area and Ventral Bed Nucleus of the Stria Terminalis Neurons that Express Fos during Maternal Behavior in Female Rats

Medial preoptic area (MPOA) and ventral bed nucleus of the stria terminalis (VBST) neurons are involved in maternal behavior, but the neural sites to which the maternally relevant neurons project have not been determined. Since MPOA and VBST neurons express Fos during maternal behavior, we used a double-labeling immunocytochemical procedure to detect both Fos and a retrograde tracer, wheat germ agglutinin (WGA), in order to determine where these Fos neurons project. On Day 4 postpartum, fully maternal females were separated from their litters. On Day 5, WGA was iontophoretically injected into one of the following regions known to receive MPOA and/or VBST input: Lateral septum, medial hypothalamus at the level of the ventromedial nucleus, lateral habenula, ventral tegmental area, retrorubral field, or periaqueductal gray. On Day 7, females received a 2-h test with either pups or candy, after which they were perfused and their brains were processed for the detection of Fos and WGA. As expected, females tested with pups had more Fos-containing neurons in the MPOA and VBST than did females tested with candy. After WGA injections into several brain sites, the number of double-labeled cells observed in the MPOA and VBST was greater for the maternal females when compared to the non-maternal females. Therefore, these results pinpointed neural circuits that were activated during maternal behavior. For the maternal females, Fos-containing neurons in the MPOA projected most strongly to the medial hypothalamus at the level of the ventromedial nucleus and to the lateral septum, while Fos-containing neurons in the VBST projected most strongly to the retrorubral field, ventral tegmental area, and medial hypothalamus. Although relatively few MPOA and VBST neurons which expressed Fos during maternal behavior projected to the periaqueductal gray, these Fos-expressing neurons made up a relatively large proportion of the MPOA and VBST projection to the periaqueductal gray. This study suggests that MPOA and VBST efferents project to a variety of regions to promote full maternal responsiveness.     

Anatomical identification of neurons in selected brain regions associated with maternal behavior deficits induced by knife cuts of the lateral hypothalamus in rats

The present experiment identified neurons associated with maternal behavior deficits induced by damage to the lateral hypothalamus (LH) in rats. Fully maternal lactating rats received bilateral coronal knife cuts through either the dorsal or ventral LH at the level of the ventromedial nucleus. The blade of the wire knife used to make the cuts was coated with horseradish peroxidase (HRP). The maternal behavior of all females was studied for 4 days postoperatively and then the brains were processed for the localization of neurons retrogradely filled with HRP. The analysis focused on those brain regions thought to be important for maternal behavior. The dorsal LH cuts severely disrupted maternal behavior while the ventral LH cuts did not. The ventral cuts labeled more medial preoptic area (MPOA) and septal-diagonal band neurons with HRP than did the dorsal cuts. The dorsal LH cuts labeled more neurons with HRP in the lateral preoptic area (LPOA), magnocellular preoptic area, bed nucleus of the stria terminalis, ventral tegmental area (VTA), substantia nigra, and central gray than did the ventral cuts. Previous research has suggested that the lateral efferents of the MPOA interact with the VTA in the control of maternal behavior. The results of the present experiment suggest that MPOA axons which descend directly to the brainstem via the ventral LH are not essential for maternal behavior. Our results are consistent with the view that an MPOA-to-LPOA-to-VTA circuit may be important for maternal behavior.     

Activation of feeding-related neural circuitry after unilateral injections of muscimol into the nucleus accumbens shell

Chemical inhibition of neurons in the nucleus accumbens shell (AcbSh) elicits intense, behaviorally specific, feeding in satiated rats. We have demonstrated previously that this treatment activates a number of brain regions, most significantly the lateral hypothalamus (LH). This activation could be elicited through a direct neural connection with the AcbSh or secondarily through changes in autonomic activity, stress, or circulating levels of orexigenic or satiety factors. In the present study, we used the immunohistochemical localization of Fos protein to map neuronal activation after unilateral muscimol injections into the AcbSh to determine whether AcbSh-mediated Fos expression remains lateralized in the circuit and whether secondary systemic changes in the rat can be excluded as primary factors in the activation of downstream component nuclei. Rats receiving only saline injections exhibited very little Fos immunoreactivity. In contrast, unilateral injections of muscimol into the AcbSh consistently increased Fos expression in several brain regions. Three distinct patterns of expression were observed. Fos synthesis in the LH was increased only on the side of the brain ipsilateral to the muscimol injection. Fos expression remained primarily ipsilateral to the injection site in the septohypothalamic, paraventricular hypothalamic (PVN), paratenial thalamic, and lateral habenular nuclei, and medial substantia nigra, but was increased bilaterally in the piriform cortex, supraoptic nucleus, central nucleus of the amygdala, and nucleus of the solitary tract. Smaller numbers of Fos-immunoreactive cells were seen unilaterally in the bed nucleus of the stria terminalis, medial ventral pallidum, arcuate nucleus, and ventral tegmental area and bilaterally in the supraoptic and tuberomammillary nuclei. The labeling in the LH, PVN, and other unilaterally labeled structures provides evidence that these brain regions are components of an AcbSh-mediated neural circuit and suggests that they may be involved in the expression of AcbSh-mediated feeding behavior.     

The influence of chemosensory input and gonadotropin releasing hormone on mating behavior circuits in male hamsters

Chemosensory input is important for mating behavior in male hamsters. Chemosignals found in female hamster vaginal fluid activate regions of the brain that receive input from the vomeronasal/accessory olfactory system and are important for mating behavior. Mating or exposure to these chemosignals produces increased Fos protein expression in the amygdala, bed nucleus of the stria terminalis, and medial preoptic area (MPOA). These brain regions contain cell bodies and/or fibers of gonadotropin releasing hormone (GnRH) neurons, suggesting potential relationships between chemosensory systems and GnRH. GnRH is released naturally when male rodents (mice and hamsters) encounter female chemosignals, and intracerebrally injected GnRH restores mating behavior in sexually naive male hamsters after removal of the vomeronasal organs. We report here that the combination of pheromone exposure and intracerebrally-injected GnRH increases Fos expression in the MPOA above the increase seen in pheromone-exposed males, or in males given only the exogenous GnRH. In males with vomeronasal organs removed (VNX), there was an also an increment in Fos expression in the MPOA when these pheromone exposed males were injected with GnRH, provided they had previous sexual experience. Males with vomeronasal organs removed and without sexual experience showed increased Fos expression in the medial amygdala when pheromone exposure and GnRH injection were combined, but not in the medial preoptic area.     

Neural basis of maternal behavior in the rat

This article presents a review of the neural and neurochemical regulation of maternal behavior in the rat, emphasizing the role of the medial preoptic area (MPOA) and its neural connections in this regulation. Evidence for the role of the MPOA includes the following and will be discussed: (1) Axon-sparing lesions of the MPOA disrupt maternal behavior, indicating the involvement of MPOA neurons rather than fibers of passage. (2) Estradiol acts on the MPOA to facilitate maternal behavior. (3) An MPOA-to-lateral preoptic area-to-ventral tegmental area circuit may be part of the output pathway by which the MPOA influences maternal behavior. (4) MPOA neural circuitry may interact with olfactory neural circuitry and with the motor system to influence maternal responsiveness. (5) Opioid neural pathways appear to inhibit, and oxytocinergic neural pathways appear to promote, maternal behavior.     

Medial preoptic area interactions with the nucleus accumbens-ventral pallidum circuit and maternal behavior in rats

Several experiments explored the roles of nucleus accumbens (NA), ventral pallidum (VP) and medial preoptic area (MPOA) in the regulation of maternal behavior in rats. A preliminary experiment found that bilateral radiofrequency lesions of medial NA did not disrupt maternal behavior. Experiment 1 found that bilateral infusions of muscimol into VP, but not into medial NA, reversibly disrupted maternal behavior. Experiment 2 found that unilateral muscimol injections into VP disrupted maternal behavior to a greater extent when paired with a contralateral N-methyl-d-aspartic acid (NMDA) MPOA lesion than when paired with a sham MPOA lesion. Experiment 3 showed that a unilateral NMDA MPOA lesion paired with a contralateral NMDA VP lesion (Contra group) disrupted maternal behavior to a much greater extent than did sham NMLA lesions or NMDA lesions of MPOA and VP ipsilateral to one another. Experiment 3 focused on the specificity of the maternal behavior disruptions and found that the primary maternal deficit in the Contra females was a severe deficit in retrieval behavior. Importantly, these females showed normal hoarding behavior, home cage activity, and elevated plus maze activity. Experiment 3 used Neu N immunohistochemistry to define the extent of MPOA and VP excitotoxic lesions. It is hypothesized that MPOA acts to facilitate the active components of maternal behavior by inhibiting NA, which then releases VP from GABAergic inhibition, and such disinhibition of VP allows pup stimuli to trigger appropriate maternal responses.     

Sexual behavior increases c-fos expression in the forebrain of the male rat.

The ability of a wide variety of pharmacological and physiological stimuli to increase neuronal expression of Fos has led to the suggestion that it might serve as a marker of neuronal activation. Psychomotor stimulants increase the release of dopamine from the terminals of nigrostriatal and mesolimbic neurons and enhance Fos immunoreactivity in the striatum and nucleus accumbens (NAc). Because sexual behavior also increases dopamine release in these and other forebrain regions, the present study examined the effect of copulation on Fos immunoreactivity in the forebrain of intact, sexually active male rats. Sexual behavior produced a striking increase in Fos immunoreactivity in the medial preoptic area (MPOA), NAc, bed nucleus of the stria terminalis and piriform cortex. However, no increase in Fos immunoreactivity was observed in the striatum. These results are consistent with neurochemical, physiological, and behavioral data suggesting that the MPOA and NAc are important substrates of sexual behavior.     

Induction of Fos-like immunoreactivity in musk shrews after mating

In many mammalian species the neuroendocrine regulation of male and female reproductive behavior is sexually dimorphic. By contrast, many features of female sexual behavior in the musk shrew (Suncus murinus) more closely resemble those of males than of females of other species. Female musk shrews require testosterone (T), which is neurally aromatized to estrogen, to induce sexual behavior. Aromatization occurs in the medial preoptic area (MPOA), and this region is critical for the expression of female receptivity. To compare neural responses to sexual behavior in females and males, we compared the number of Fos-like immunoreactive (Fos-ir) neurons after mating in musk shrews. In both males and females the number of Fos-ir neurons was increased by mating activity in the granule layer of the accessory olfactory bulb (gr-AOB), the bed nucleus of the stria terminalis (BNST), MPOA, the medial amygdala (MeA), and the region corresponding to the midbrain central tegmental field (CTF). Although Fos was induced by mating in several regions, this response was only dimorphic in the ventral medial nucleus of the hypothalamus (VMN), where mating significantly increased Fos-ir in females, but not in males. In both sexes, only the gr-AOB displayed an increase in Fos-ir after exposure to chemosensory cues alone. Thus, the pattern of Fos expression in the brain after mating is only sexually dimorphic in one region, the VMN. Further, in spite of past behavioral studies done in this species, which show a role for pheromones in induction of receptivity, these data show that exposure to pheromones does not induce Fos in structures caudal to the olfactory bulbs.     

Anatomical interrelationships of the medial preoptic area and other brain regions activated following male sexual behavior: A combined Fos and tract-tracing study

The medial preoptic nucleus (MPN) is an essential site for the regulation of male sexual behavior. Previous studies using c-fos as a marker for neural activation have shown that copulation increased c-fos expression in the MPN. Neural activation was also present in brain regions that are connected with the MPN and are involved in male sexual behavior, including the posteromedial bed nucleus of the stria terminalis (BNSTpm), posterodorsal preoptic nucleus (PD), posterodorsal medial amygdala (MEApd), and parvocellular subparafascicular thalamic nucleus (SPFp). The present study investigated whether the copulation-induced, activated neurons in these brain regions are involved in the bidirectional connections with the MPN. Therefore, mating-induced Fos expression was combined with application of anterograde (biotinylated dextran amine) or retrograde (cholera toxin B subunit) tracers in the MPN. The results demonstrated that neurons in the BNSTpm, PD, MEApd, and SPFp that project to the MPN were activated following copulation. However, in males that displayed sexual behavior but did not achieve ejaculation, few double-labeled neurons were evident, although both retrogradely labeled neurons and Fos-immunoreactive cells were present. In addition, retrograde neurons that expressed Fos were located in discrete subdivisions within the brain regions studied, where Fos is induced after ejaculation. Likewise, anterogradely labeled fibers originating from the MPN were not distributed homogeneously but were particularly dense in these discrete subdivisions. These results demonstrate that copulation-induced Fos-positive neurons in specific subdivisions of the BNSTpm, PD, MEApd, and SPFp have bidirectional connections with the MPN. Taken together with previous findings, this supports the existence of a discrete subcircuit within a larger neural network underlying male sexual behavior. J. Comp. Neurol. 397:421–435, 1998. © 1998 Wiley-Liss, Inc.     

Effects of discrete lesions of the sexually dimorphic nucleus of the preoptic area or other medial preoptic regions on the sexual behavior of male rats

The sexually dimorphic nucleus of the rat medial preoptic area (SDN-POA) has a volume five times larger in the adult male compared with that of the adult female. In the present study, the effects of discrete electrolytic destruction of the SDN-POA or other specific medial preoptic (MPOA) regions on masculine sexual behavior were determined in adult, sexually experienced male rats. Small lesions encompassing the SDN-POA had no effect on the maintenance of copulatory behavior. Lesions of similar size placed within the ventral or anterio-dorsal MPOA also did not consistently affect the display of masculine sexual behavior. However, animals that received small lesions within their dorsal MPOA showed a substantial, long-term decrease in number of mounts, intromissions, and ejaculations compared to these parameters in sham-lesioned control rats, thus indicating a lesion-induced disruption of those neural mechanisms mediating these behaviors. Collectively these data suggest that the SDN-POA is not critical for a full expression of male sexual behavior and that the dorsal MPOA may be more important than other MPOA regions for copulatory behavior.     

Neonatal manipulation of oxytocin influences the partner preference in mandarin voles (Microtus mandarinus)

Neonatal manipulation of oxytocin (OT) has long-term effects on behavior and physiology. The objective of this research was to determine if neonatal exposure to OT can affect partner preferences and to characterize the mechanisms underlying social behavior such as neural activities of relevant brain regions in socially monogamous mandarin voles (Microtus mandarinus). After receiving a subcutaneous injection of isotonic saline (SAL) or OT within 24h of birth, mandarin vole at 60 days of age was paired with an unfamiliar opposite sex for 24h, followed immediately by an examination of their partner preference and 30 days later by the second examination of their partner preference and Fos expression in some brain regions. The results indicated that (1) while 24h cohabitation was insufficient for both female and male SAL-treated mandarin voles to form partner preference, neonatal exposure to OT significantly facilitate female, but not male mandarin voles to form partner preference within 24h of cohabitation; (2) the maintenance of partner preference in females was suppressed by neonatal OT treatment, while neonatal OT-treated males showed significant partner preference as neonatal SAL-treated males and females; in addition, the tendency of aggression to the strangers was impaired in both females and males, and neonatal OT-treated males showed significantly higher mounting behavior to the partner; (3) in comparison with saline-treated females, OT-treated females showed a significant decrease in Fos expression in all brain regions examined in response to partner preference. Relative to saline treatment, neonatal OT treatment induced to males a significant decrease of Fos expression in the bed nucleus of the stria terminalis (BNST), the hypothalamic paraventricular nucleus (PVN) and the mediodorsal thalamic nucleus (MD) as well as a significant increase in the medial preoptic area (MPOA), the dorsal part of the lateral septal nucleus (LSD) and the central amygdaloid nucleus (CeA). These results demonstrate that neonatal OT exposure has different effects on the formation and maintenance of partner preference in mandarin voles and the latter effects possibly via OT-induced changes of the neural activities of relevant brain regions.     

Differences in Fos expression in the rat brains between cold and warm ambient exposures

Fos expression in the rat diencephalon, brain stem, cerebellum, and spinal cord was examined after warm (33°C) and cold (10°C) ambient exposures. Fos expression was examined with use of immunohistochemical method and the number of Fos-positive neurons In each nucleus was quantitatively analyzed. When rats were exposed to cold ambient, significant number of Fos-positive neurons was found in the lateral septal nucleus (LS), preoptic hypothalamic area (POA), parvocallular paraventricular hypothalamic nucleus (PPVN), lateral preoptic area (1-130), zone incerta (ZI), pareventricular thalamic nucleus (PV), ventromedial hypothalamic nucleus (VMH), subparafesclcutm thalamic nucleus (SPF), postrior hypothalamic area (PH), supramammillary nucleus (SuM), microcellular tegmental nucleus (MiTg), lateral iemomscus nucleus (LL), lateral dorsal central grey (CGLD), lateral ventral central grey (CGLV), dorsal parabrachial nucleus (DPS), locus coeruleus (LC), dorsal tegmental nucleus (DTg), vestibular nucleus (Ves), nucleus of solitary tract (Sol), spinal cord, and cerebellum. When animals were exposed to warm ambient, the numbers of Fos-positive neurons in the LS, POA, PV, LPO, and SuM were significantly increased to be equal to those of cold ambient However, after warm ambient exposure the numbers of Fos-positive neurons in the DPB and spinal cord were increased but less than those of cold ambient, and those in the pPVN, VMH, ZI, SPF, PH, CGLD, CGLV, Mg, LL, LC, DTg, Ves, Sol, and cerebellum were not significantly increased as compared with those of control or cold ambient Abdominal temperature was not changed doing cold ambient exposure, but the temperature was sWNcantly increased during warn ambient exposure. These results demonstrate that Fos-positive regions in the rat brains are sagraficantly different between cold and worm ambient exposures although several hypothalamic brain regions reveal the same degree of Fos expression, irxiCating the differences of neural patiwways concerning thennoregulation between cold and warm ambients.     

Effects of transecting lateral neural connections of the medial preoptic area on maternal behavior in the rat: Nest building, pup retrieval and prolactin secretion

In lactating female rats bilateral parasagittal cuts transecting the dorsolateral neural connections of the medial preoptic area (MPOA) abolished nest building and retrieving components of maternal behavior, while crouching and nursing were unaffected. While few animals with these cuts were suckled when presented with pups the prolactin secretion response was undiminished when suckling did occur. Whereas previous studies have demonstrated that extensive surgical separation of the medial preoptic area-anterior hypothalamic continuum from the lateral preoptic area-lateral hypothalamus disrupts all aspects of maternal behavior, the present study has determined more specifically the zone of fibers essential for the active components of maternal behavior, i.e., nest building and retrieving. These fibers appear to enter/leave MPO dorsolaterally beneath the crossing of the anterior commissure in the region of the bed nucleus of the stria terminalis. These neural connections are not the same as those that regulate prolactin secretion in the lactating rat.     

Colocalization of androgen receptors and mating-induced FOS immunoreactivity in neurons that project to the central tegmental field in male rats.

Bilateral lesions of the central tegmental field (CTF) in male rats virtually eliminate mating behavior. This study examined if mating-induced Fos expression (a measure of neuronal activation) and androgen receptors (AR) are colocalized in brain and spinal cord neurons which project to the CTF. Animals received unilateral injections of the retrograde tracer Fluorogold (FG) in the lateral part of the CTF (CTFl), and 10 days later were killed after ejaculating with females. Brains and spinal cords were examined for FG transport, AR-immunoreactivity (AR-ir), and Fos-immunoreactivity (Fos-ir). AR-ir and Fos-ir were visualized with fluorescence microscopy using cyanine-conjugated and fluorescein-conjugated secondary antibodies. The CTFl received projections from AR-containing neurons in forebrain structures (bed nucleus of stria terminalis, medial preoptic area, lateral and ventromedial hypothalamus), in the central amygdala and various mid- and hindbrain structures (dorsolateral tegmentum, superior and inferior colliculi, pedunculopontine nucleus), and in the lumbosacral spinal cord (lamina X). Some of the AR-containing neurons in bed nucleus of stria terminalis and in the dorsal part of the medial preoptic area with projections to the CTFl were activated by mating. Most AR-containing neurons in spinal lamina X with projections to the CTFl were also activated by mating. Information from spinal cord and pontine nuclei and from outputs descending from the forebrain may be relayed in the CTFl. Thus, as part of a network of hormone-sensitive neurons linking brain and spinal cord mechanisms for mating, the CTFl could participate in the integration of visceral and somatic information relevant for sexual behavior.     

Neural steroid hormone receptor gene expression in pregnant rats

Estrogen and progesterone play important roles during pregnancy in stimulating the onset of maternal behavior at parturition. The status of receptor expression of these hormones during pregnancy in neural regions that regulate maternal behavior is unclear. The objective of the present study is to characterize changes in neural gene expression of the estrogen receptors alpha and beta (ERalpha and ERbeta) and the progesterone receptor (PR) during the latter part of pregnancy. Brains from primigravid Sprague-Dawley rats were collected on days 15 and 21 of pregnancy. Micropunches of the olfactory bulb (OB), medial preoptic area (MPOA), bed nucleus of the stria terminalis (BnST), hypothalamus (HYP), medial amygdala (MeA), and the temporal cortex (TCx) were analyzed by real-time RT-PCR (Taqmantrade mark) for levels of gene expression. No changes in either ERalpha or ERbeta mRNA levels were detected in any brain region between days 15 and 21 of pregnancy: however, the MPOA had higher levels of both ERalpha and ERbeta than other brain regions. Progesterone receptor mRNA levels, in contrast, declined significantly in the MPOA, HYP, and TCx, between days 15 and 21 of pregnancy (P < 0.05). In addition, the levels of PR mRNA were significantly higher in the HYP and TCx compared to both the OB and MeA. These data indicate that there is a downregulation of PR prepartum and suggest that this decrease may play a role in the disinhibition of maternal behavior at parturition.     

Prolactin receptor gene expression in the forebrain of pregnant and lactating rats

Prolactin plays a large role in the onset of maternal behavior at parturition. Knowledge of the change in expression of the prolactin receptor in the brain across pregnancy and lactation, however, is limited. Prolactin receptor gene expression was determined by in situ hybridization histochemistry during pregnancy and lactation in rats. Expression of the mRNA for the longform of the prolactin receptor (PRL-R-L) was measured in various forebrain structures in primigravid rats at different stages of pregnancy, in primiparous rats during early, mid-, and late lactation, and in age-matched, nulliparous females in diestrus. Hybridizations were performed using a [33P]-labeled riboprobe specific for the long form of the prolactin receptor mRNA complimentary to 290 bp of the prolactin receptor gene. The following areas of the forebrain were examined: medial preoptic area (MPOA), median preoptic nucleus both dorsal (MePOd) and ventral (MePOv) to the anterior commissure, ventral lateral septum (LSv), and the ventral and principal parts of the bed nucleus of the stria terminalis (BnSTv and BnSTpr, respectively). Overall, the number of cells expressing PRL-R-L mRNA was significantly higher at 2 h postpartum compared to diestrus in all areas examined except the LSv. In addition, there were lower numbers of PRL-R-L cells during all stages of lactation compared to pregnancy. The number of grains per cell in the MPOA and LSv did not change as dramatically as the number of cells expressing PRL-R-L mRNA in those brain regions. These data contribute to the growing body of evidence that the neural lactogenic system changes as a function of female reproductive state. Changes in PRL-R-L mRNA in terms of behavior and endocrine functions are discussed.     

Metabolic mapping of the brain in pregnant, parturient and lactating rats using Fos immunohistochemistry

The present study was designed to investigate Fos-positive neurons of the female rat brain at various reproductive states in order to analyze the metabolic map connected with pregnancy, parturition and lactation. The number of Fos-positive neurons in each brain nucleus was analyzed with a quantitative immunohistochemical method in virgin, pregnant, parturient, lactating and arrested lactating rats. In parturient rats, a significant number of Fos-positive neurons was observed as compared to virgin or pregnant females in the following brain regions; the bed nucleus of the stria terminalis (BST), lateral septal nucleus (LS), medial preoptic area (MPA), periventricular hypothalamic nucleus (Pe), parvocellular paraventricular hypothalamic nucleus (PaPVN), magnocellular paraventricular hypothalamic nucleus (MaPVN), supraoptic nucleus (SON), paraventricular thalamic nucleus (PV), anterior hypothalamic area (AHA), lateral hypothalamic area (LH), amygdaloid nucleus (AM), supramammillary nucleus (SuM), substantia nigra (SN), central grey (CG), microcellular tegmental nucleus (MiTg), subparafascicular thalamic nucleus (SPF), posterior hypothalamic area (PH), dorsal raphe nucleus (DR), locus coeruleus (LC), dorsal parabrachial nucleus (DPB), nucleus of solitary tract (Sol), and ventrolateral medulla (VLM). Significant differences were found in the number of Fos-positive neurons between parturient and lactating females, although localization of Fos-positive neurons in lactating females was quite similar to parturient ones. Between parturient and lactating rats: (1) In the MPA, PaPVN, AHA, arcuate hypothalamic nucleus (Arc), ventromedial hypothalamic nucleus (VMH), mesencephalic lateral tegmentum (MLT), and genual nucleus (Ge), the number of Fos-positive neurons of lactating females were significantly higher than those of parturient ones; (2) In the LS, Pe, PV, LH, AM, SuM, CG, MiTg, SPF, PH, DR, LC, and VLM, there was no significant differences in the number of Fos-positive neurons; (3) In the BST, MaPVN, SON, SN, DPB and Sol, the number of Fos-positive neurons of lactating rats were significantly lower than those of parturient ones. These different patterns of Fos expression among many brain regions may be owing to the functional differences in each region. Fos expression in lactating rats was apparently induced by suckling stimulation because the removal of their litters immediately after parturition completely eliminated expression of Fos protein in each nucleus. These results suggest that the localization of Fos-positive neurons in a number of neural populations throughout the brain may be revealing the neural circuits in response to parturition or lactation.     

Chronic tooth pulp inflammation causes transient and persistent expression of Fos in dynorphin-rich regions of rat brainstem

We have analyzed central Fos immunoreactivity (Fos-IR) brainstems of adult rats after three clinically relevant dental injuries: filled dentin (DF) cavities that cause mild pulp injury and heal within 1-2 weeks; open pulp exposures (PX) that cause gradual pulp loss and subsequent periodontal lesions; and filled pulp exposures (PXF). By 1 week after DF cavities, no Fos-IR remained except for sites such as lateral-ventral periolivary nucleus (LVPO) that had Fos-IR in all rats including controls. PX injury induced (1) a delayed transient expression of Fos at 1-2 weeks at three loci (ipsilateral neurons in dorsomedial nucleus oralis, paratrigeminal nucleus, and trigeminal tract), (2) persistent ipsilateral Fos for at least 4 weeks after injury in dynorphin (Dyn)-rich regions (rostral lateral solitary nucleus, periobex dorsal nucleus caudalis), and (3) late Fos-IR at 2-4 weeks (bilateral superficial cervical dorsal horn, contralateral dorsal nucleus caudalis, contralateral rostral lateral solitary nucleus). Rats with PXF injury were examined at 2 weeks, and they had greater numbers and more extensive rostro-caudal distribution of Fos neurons than the PX group. One week after PX injury, Fos-IR neurons were found in regions with strong Dyn-IR central fibers. Co-expression of Dyn and Fos was found in some unusually large neurons of the ipsilateral rostral lateral solitary nucleus, trigeminal tract, and dorsal nucleus caudalis. Immunocytochemistry for the p75 low affinity neurotrophin receptor (p75NTR) or for calcitonin gene-related peptide (CGRP) showed no consistent change in trigeminal central endings in any Fos-reactive brainstem areas, despite the extensive structural and cytochemical reorganization of the peripheral endings of the dental neurons. The Fos responses of central neurons to tooth injury have some unusual temporal and spatial patterns in adult rats compared to other trigeminal injury models.     

Efferent projections from the region of the medial zona incerta containing A13 dopaminergic neurons: a PHA-L anterograde tract-tracing study in the rat

Potential efferent projections of A₁₃ dopaminergic (DA) neurons were identified in the present study by examining the distribution of labelled fibers following iontophoretic injection of the anterogradely transported lectinPhaseolus vulgaris leucoagglutinin (PHA-L) into the medial zona incerta (MZI), the region of the diencephalon containing A₁₃ DA neuronal perikarya. One week after injection, PHA-L labelled fibers were found throughout the brain with the heaviest labelling occurring ipsilateral to the injection site in the anterior hypothalamic area, lateral hypothalamus, lateral preoptic area, horizontal diagonal band of Broca, and parvocellular region of the paraventricular nucleus. Moderate labelling was observed in the ipsilateral median preoptic nucleus, lateral septum, lateral aspect of the bed nucleus of the stria terminalis, and central nucleus of the amygdala. Moderate labelling was also found in the contralateral MZI and parvocellular region of the paraventricular nucleus. Light labelling was detected in the ipsilateral medial preoptic area, supraoptic nucleus, ventromedial nucleus, arcuate nucleus, vertical limb of the diagonal band of Broca, and in the contralateral lateral hypothalamus. Few immunopositive fibers were present in the dorsomedial nucleus of the hypothalamus or the magnocellular region of the paraventricular nucleus. These results reveal that neurons located in the MZI (possibly A₁₃ DA neurons) have ipsilateral efferent axonal projections to a variety of brain regions including the lateral hypothalamus, lateral preoptic area, and the limbic structures at the diencephalic-telencephalic juncture.