Expression of Tumour Endothelial Marker 8 in Canine Mammary Gland Tumour Cells.

The aim of this study was to investigate the expression of tumour endothelial marker 8 (TEM8) in canine mammary gland tumours (MGTs) by immunohistochemistry and to evaluate the association between tumour cell TEM8 expression and tumour histological features, histological grades and expression of luminal and basal/myoepithelial cell markers. TEM8 expression was detected in >60 % of neoplastic epithelial cells in all simple adenomas (n = 25), simple carcinomas (n = 43) and invasive micropapillary carcinomas (n = 5) studied. Six of the 18 solid carcinomas studied showed TEM8 expression in >60% of carcinoma cells present in solid structures and in 12 of the 18 solid carcinomas, <30% of the luminal structure-forming carcinoma cells showed TEM8 expression. TEM8 expression in the neoplastic cells was not associated with histological malignancy in canine MGTs. TEM8+ tumour cells frequently showed the luminal-like phenotype cytokeratin (CK)19+/p63-/α-smooth muscle actin (SMA)-, while most TEM8- tumour cells exhibited the basal-like phenotype CK19-/p63+/αSMA-. These findings indicate that TEM8 may be involved in maintaining the characteristics of luminal cells in canine MGTs and that TEM8 would be useful in identifying the type of neoplastic epithelial cell in MGTs.

Two routes for pollen entering indoors: ventilation and clothes.

BACKGROUND: The route by which pollen enters dwellings has not been clarified. OBJECTIVE: To evaluate the amount of pollen entering dwellings by ventilation and adhesion to textile products. METHODS: The amount of pollen clinging to fabrics (clothes, laundry, and futon bedding) out of doors was measured by quantification of Japanese cedar pollen antigen Cry j 1. The effect of air ventilation on the amount of pollen indoors was also investigated using several neighboring unoccupied apartments with an identical layout while controlling the ventilation conditions. RESULTS: The amount of pollen adhering to futons was especially high. More than half of the pollen on futons or laundry remained on the surface, even after being brushed off by hand or shaken off. Vacuuming laundry and futons after airing out would be an effective way to decrease the amount of indoor pollen. A large amount of pollen entered dwellings through air ducts when the windows were closed and the ventilation fans working. Since most pollen that entered by ventilation remained near the windows, cleaning carefully and frequently near windows could reduce the amount of pollen indoors. CONCLUSIONS: To decrease the amount of pollen indoors, special attention must be paid to textile products and ventilation systems during the pollen season.

Circadian rhythm of melatonin release from the photoreceptive pineal organ of a teleost, ayu (Plecoglossus altivelis) in flow-thorough culture.

In the present study, we tested whether the pineal organ of ayu (Plecoglossus altivelis), an osmerid teleost close relative of salmonids, harbours a circadian oscillator regulating rhythmic melatonin release using flow-through culture. The pineal organ maintained under light/dark cycles released melatonin in a rhythmic fashion with high levels during the dark phase. A circadian rhythm of melatonin release persisted in constant darkness for at least four cycles. Characteristics of the circadian rhythm (free-running period, phase and amplitude) exhibited small variations among cultures when the data was normalized, indicating that this system is sufficient for the analysis of the circadian rhythm both at qualitative and quantitative levels. Six-hour extension of the light phase from the normal onset time of the dark phase or exposure to constant light for 36 or 48 h before transfer to constant darkness significantly inhibited melatonin release. Phase shifts in the circadian rhythm of melatonin release were also observed. Thus, the ayu pineal organ contains all the three essential components of the circadian system (a circadian clock, the photoreceptor responsible for photic entrainment of the clock, and melatonin generating system as an output pathway). This system should provide a useful model for analysing the physiological and molecular basis of the vertebrate circadian system. In addition, further comparative studies using salmonids and related species including ayu will provide some insight into the evolution of the roles of the pineal organ in the vertebrate circadian system.

Transfer of canine embryos at various developmental stages recovered by hysterectomy or surgical uterine flushing.

In dogs, embryo transfer (ET) techniques such as induciton of excessive ovulation and synchronization of estrus have not progressed well. Therefore, using embryos at various developmental stages, ET was investigated in dogs from a beagle colony in which the ovulation days were close, as estimated by the progesterone level. Embryos were, recovered 8-11 days after ovulation (4-9 days after mating) by excising the oviducts and uteri (excision method) in 16 animals and by surgical flushing of the uteri at laparotomy (surgical method) in 3 animals. In 24 dogs with -4 to +2 days of difference in the timing of ovulation between donor and recipient dogs, 1-10 embryos at the 8-cell to blastocyst stages were transferred per animal. The mean embryo recovery rate by the excision method (97.1%) was significantly higher than that by the surgical method (42.5%) (p<0.01). Twelve (57.1%) of 21 animals with -1 to +2 days difference in ovulation day became pregnant after the transfer of 8-cell to blastocyst stage embryos. Although 3 dogs with -4 to -2 days of difference of ovulation day underwent ET of morula or compacted morula, none of these dogs became pregnant. The mean ratio of the number of newborns to the number of transferred embryos was only 51.9%. The mean duration of the period between ovulation and delivery in the pregnant recipients was 65.8 days, which tended to be longer than that in natural mating. These results demonstrate that pregnancy can be induced by ET at the 8-cell to blastocyst stage in dogs with -1 to +2 days difference in ovulation day.

Melatonin, a pineal secretory product with antioxidant properties, protects against cisplatin-induced nephrotoxicity in rats.

In an attempt to define the role of the pineal secretory melatonin and an analogue, 6-hydroxymelatonin (6-OHM), in limiting oxidative stress, the present study investigated the cisplatin (CP)-induced alteration in the renal antioxidant system and nephroprotection with the two indolamines. Melatonin (5 mg/kg), 6-OHM (5 mg/kg), or an equal volume of saline were administered intraperitoneally (i.p.) to male Sprague Dawley rats 30 min prior to an i.p. injection of CP (7 mg/kg). After CP treatment, the animals each received indolamine or saline every day and were sacrificed 3 or 5 days later and plasma as well as kidney were collected. Both plasma creatinine and blood urea nitrogen increased significantly following CP administration alone; these values decreased significantly with melatonin co-treatment of CP-treated rats. In the kidney, CP decreased the levels of GSH (reduced glutathione)/GSSG (oxidized glutathione) ratio, an index directly related to oxidative stress. When animals were treated with melatonin, the reduction in the GSH/GSSG ratio was prevented. Treatment of CP-enhanced lipid peroxidation in the kidney was again prevented in animals treated with melatonin. The activity of the antioxidant enzyme, glutathione peroxidase (GSH-Px), decreased as a result of CP administration, which was restored to control levels with melatonin co-treatment. Upon histological analysis, damage to the proximal tubular cells was seen in the kidneys of CP-treated rats; these changes were prevented by melatonin treatment. 6-OHM has been shown to have some antioxidative capacity, however, the protective effects of 6-OHM against CP-induced nephrotoxicity were less than those of melatonin. The residual platinum concentration in the kidney of melatonin co-treated rats was significantly lower than that of rats treated with CP alone. It is concluded that administration of CP imposes a severe oxidative stress to renal tissue and melatonin confers protection against the oxidative damage associated with CP. This mechanism may be reasonably attributed to its radical scavenging activity, to its GSH-Px activating property, and/or to its regulatory activity for renal function.

The mPVN mediates blockade of NPY-induced feeding by a Y5 receptor antagonist: a c-FOS analysis.

To identify the site(s) of NPY Y5 receptor (Y5R) mediation of NPY-induced feeding, we employed c-Fos immunostaining and a selective Y5R antagonist (Y5R-A), CGP71683A, in adult male rats. Intracerebroventricular (icv) administration of NPY stimulated feeding and c-Fos-like immunoreactivity (FLI) in the dorsomedial hypothalamus, supraoptic nucleus and the two subdivision of the hypothalamic paraventricular nucleus (pPVN), the parvocellular (pPVN), and magnocellular (mPVN). Y5R-A on its own, injected either intraperitoneally or icv, neither affected feeding nor FLI in hypothalamic sites. However, Y5R-A pretreatment suppressed NPY-induced food intake and FLI selectively in the mPVN. Taken together with our previous similar finding of Y1R involvement, these results suggest that NPY receptor sites concerned with feeding behavior reside selectively in the mPVN and Y1 and Y5 receptors are either coexpressed or expressed separately in those target neurons that promote appetitive drive.

Measurement of the adhesive force of fine particles on tablet surfaces and method of their removal.

The adhesion force of fine particles on the surface of tablets was measured by a centrifugal force and impact separation method. A Finededuster (FDD) was employed to remove fine particles from the tablet surface. The centrifugal force and impact separation method was suggested to be effective for measuring the adhesive forces between particles and the tablet surface, and effective disjoining force in the FDD could be estimated by comparison of the results obtained using these two methods. The FDD showed high removal efficiency regardless of how many tablets were processed at the same time. In either of these methods, critical particle size was about 10-20 microns, and larger particles were removed more efficiently. This critical particle size was similar to that observed for other mechanical properties of powders, such as angle of repose and flowability. We simulated particle residual percentage under various operation conditions by ANN (artificial neural network) analysis and multiple regression analysis. This simulation enabled us to predict how the efficiency of particle removal is affected by the interaction of the experimental and material factors.

Proteins recognized by antibodies against isolated cytological heterochromatin from rat liver cells change their localization between cell species and between stages of mitosis (interphase vs metaphase).

Heterochromatin in the cell nucleus seems to concentrate various proteins, such as Drosophila heterochromatin protein 1, which maintain the repressed state of gene expression. However, it still remains obscure how protein composition related to chromatin structure is different between heterochromatin and euchromatin in interphase nuclei. We isolated cytological heterochromatin from sonicated interphase nuclei obtained from rat liver cells and prepared antisera against it. The dense heterochromatic bodies seen in the preparation of intact nuclei were duplicated in a relatively pure form during the preparation of heterochromatin. In the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, differences between the fractions of heterochromatin and euchromatin were noted by their protein composition. Isolated heterochromatin was then digested by DNase after partial digestion with trypsin and its dense structure changed to become highly sensitive to DNase. The prepared antibodies reacted with the heterochromatin region of rat liver cell nuclei and isolated cytological heterochromatin; however, they did not react with euchromatin. Using immunohistochemistry, the antibodies bound to each cell nucleus in all tissues observed; some cell types were distinguished by their differential stainability (e.g. staining in the cytoplasm). Staining of the mitotic cells showed that the proteins recognized by the antibodies were localized in the cytoplasm and, in part, on the chromosomes. Based on the results of molecular cloning from rat liver cDNA library using the antibodies as a probe, it seemed that the antibodies mainly recognized two proteins similar to arginase and general vesicular transport factor p115, respectively. The results obtained from these experiments reveal that some proteins located in the heterochromatin of interphase liver cell nuclei seem to play important roles in condensing a portion of the chromatin structure during interphase and suggest that proteins composing heterochromatin might be changed according to cell types or the stage of the cell cycle.

Specific expression pattern of Fos in the accessory olfactory bulb of male mice after exposure to soiled bedding of females.

The heterogeneous structure of the accessory olfactory bulb (AOB) has been demonstrated immunocytochemically. In this study, we analyzed the expression of an immediate-early gene protein, c-Fos, as a marker of neuronal activity in response to chemosensory cues was analyzed. The number of c-Fos-immunoreactive (Fos-ir) cells was measured in the rostral and caudal zones of the AOB in male ICR mice after exposure to the soiled bedding of female mice. The results revealed no significant difference in the number of Fos-ir cells in the caudal zone of the AOB between exposure to the soiled bedding of female ICR mice (ICR group) and exposure to that of female Balb mice (Balb group). In the rostral zone, however, the number of Fos-ir cells in the glomerular layer and granule cell layer was larger in the ICR group than in the Balb group. The difference in the expression of c-Fos in response to different pheromonal stimuli between the rostral and caudal zones in the mouse AOB has been shown for the first time in this study. These results strongly suggest that the heterogeneous structure of the AOB has an important role in the perception and processing of pheromones.

Inhibition of neuropeptide Y (NPY)-induced feeding and c-Fos response in magnocellular paraventricular nucleus by a NPY receptor antagonist: a site of NPY action.

Neuropeptide Y (NPY) is one of the important endogenous orexigenic peptides. In these studies we employed c-Fos immunostaining and a selective NPY Y1 receptor antagonist to identify the site of action of NPY in the hypothalamus. The results showed that intracerebroventricular administration of NPY stimulated feeding and increased immunostaining of c-Fos, a product of the immediate early gene c-fos, in several hypothalamic sites, including the dorsomedial nucleus, the supraoptic nucleus, and the two subdivisions of the paraventricular nucleus (PVN), the parvocellular PVN, and magnocellular PVN (mPVN). Intracerebroventricular administration of 1229U91, a selective NPY Y1 receptor antagonist, affected neither food intake nor c-Fos-like immunoreactivity (FLI) in these hypothalamic sites. Co-administration of NPY and NPY Y1 receptor antagonist inhibited NPY-induced food intake by 48%, but failed to affect NPY-induced FLI in the supraoptic nucleus, dorsomedial nucleus, and parvocellular PVN. However, this combined treatment decreased FLI by 46% in the mPVN (P < 0.05). These results showed that whereas NPY can stimulate FLI in several hypothalamic sites, the selective NPY Y1 antagonist suppressed NPY-induced FLI only in the mPVN. Thus, these findings lend credence to the view that a subpopulation of Y1 receptor-containing neurons in the mPVN in part mediate stimulation of feeding by NPY.

Female-soiled bedding induced fos immunoreactivity in the ventral part of the premammillary nucleus (PMv) of the male mouse.

Previous studies have indicated that the ventral part of the premammillary nucleus (PMv) of rodents is involved in the regulation of aggressive and male mating behavior, although the precise physiological function of the PMv is still unclear. To analyze the physiological role of the PMv in male mating behavior, the effects of exposure to bedding soiled by female mice on Fos immunoreactivity (Fos-ir), an early marker of neuronal activation, were studied in the PMv and some sex-related nuclei. We observed that exposure to female-soiled bedding induced Fos-ir expression in the PMv of the male mouse. Although Fos-ir positive cells were found in the posterodorsal part of the medial amygdaloid nucleus and in the posteromedial cortical amygdaloid nucleus, which are terminals of the neuronal projections from the main and accessory olfactory bulbs, the numbers of Fos-ir cells in those nuclei were not affected by exposure to female-soiled bedding. Moreover, Fos-ir was not detected in the ventromedial hypothalamic nucleus. It is well established that soiled bedding is useful as a source of chemosensory substances, which include "pheromones." Thus, our findings, in agreement with previous behavioral and anatomical data, suggest that the PMv plays a role in initiating male copulative behavior that is induced by a female mice pheromone(s).

Neural substrates for leptin and neuropeptide Y (NPY) interaction: hypothalamic sites associated with inhibition of NPY-induced food intake.

Intracerebroventricular (i.c.v.) injection of leptin, the adipocyte hormone, inhibits neuropeptide Y (NPY)-induced feeding in the rat. To identify the neural substrate for leptin and NPY interaction in the hypothalamus, we evaluated the expression of c-fos-like immunoreactivity (FLI), an early marker of neuronal activation, in response to icv administration of leptin, NPY and leptin plus NPY. As expected, leptin significantly decreased NPY-induced feeding in leptin plus NPY-treated rats. A comparative evaluation of the number of FLI-positive neurons in hypothalamic sites showed that both leptin and NPY activated FLI in the parvocellular subdivision of the paraventricular nucleus (pPVN), dorsomedial nucleus (DMN) and ventromedial nucleus (VMN). NPY also augmented the FLI response in the magnocellular PVN (mPVN) and supraoptic nucleus (SON), two sites where leptin alone was ineffective. Combined leptin and NPY treatment significantly decreased the number of FLI-positive neurons in the magnocellular PVN but increased their number in the dorsomedial nucleus as compared to the number of FLI-expressing neurons in these sites after NPY and leptin alone. Because there is morphologic evidence of a link between magnocellular PVN and dorsomedial nucleus, these results suggest the functional involvement of leptin plus NPY responsive elements in these sites in reduction of NPY-induced feeding by leptin.

Glial and neuronal localization of neuronal nitric oxide synthase immunoreactivity in the median eminence of female rats.

Localization of neuronal nitric oxide synthase-immunoreactivity (nNOS-IR) in the median eminence of female rats (n=4) was examined by electron microscopy to explore the possibility that nitric oxide is involved in the terminal regulation of neurosecretory peptides such as GnRH. Under light microscopy, a dense distribution of nNOS-IR was observed in this region. Electronmicroscopically, nNOS-IR was found in glial elements and nerve terminals containing dense-core vesicles. We also found a few nNOS-immunopositive synapses, in which intense immunoreactivity was found on the postsynaptic density and mitochondrial membrane. The localization of nNOS-IR in nerve terminals and glial elements in the median eminence might indicate that nNOS plays a role in regulating the release of neurosecretory peptide.

Postnatal development and sex difference in neurons containing estrogen receptor-alpha immunoreactivity in the preoptic brain, the diencephalon, and the amygdala in the rat.

Estrogen has been considered as a key substance that induces sexual differentiation of the brain during fetal and neonatal life in the rat. Thus, to define the brain regions involved in the brain sexual differentiation, we examined the regions where the estrogen receptor (ER) is located in the developing rat brain. We examined immunohistochemical distribution of the cells containing estrogen receptor-alpha (ER-alpha) in the preoptic region, the diencephalon, and the amygdala in male and female rats on postnatal days 1-35 (PD1-PD35). The antibody used recognizes ER-alpha equally well for both occupied and unoccupied forms. ER-alpha immunostaining was restricted to the cell nuclei of specific cell groups. In PD1 rats, ER-alpha-immunoreactive (ER-IR) signals were detected in the lateral septum, the organum vasculosum lamina terminalis, the medial preoptic nucleus (MPN), the median preoptic nucleus, the bed nucleus of the stria terminalis, the hypothalamic periventricular nucleus, the lateral habenula, the posterodorsal part of the medial amygdala nucleus, the posterior part of the cortical amygdala nucleus, the hypothalamic ventromedial nucleus (VMH), the hypothalamic arcuate nucleus, and the posterior hypothalamic periventricular nucleus. The distribution pattern of ER-IR cells in the newborn rat was much the same as that in the adult in the preoptic-hypothalamic and amygdala regions. Moreover, the signals in the MPN and the VMH were stronger in the female than in the male, perhaps reflecting the ability ofestrogen generated by aromatization of testosterone in the male to down-regulate the ER signal. Thus, the brain regions showing sex differences may be sites of sexual differentiation of the brain by aromatizable androgen during the neonatal period.

Co-localization of androgen receptor and nitric oxide synthase in the ventral premammillary nucleus of the newborn rat: an immunohistochemical study.

The distribution of the neuronal nitric oxide synthase (nNOS), androgen receptor (AR), estrogen receptor (ER) and aromatase (ARO) was studied in the dorsal and ventral premammillary nuclei (PMd and PMv) of the newborn rat by immunohistochemistry. In the intact male pups, nNOS immunoreactivity (-IR) was present both in the PMd and the PMv, while AR-IR was detected only in the PMv. On the other hand, ER-IR and ARO-IR were scarcely encountered in the both PMd and PMv. By double immunostaining of nNOS and AR, all the nNOS-IR cells in the PMv were revealed to contain AR-IR. In the intact female pups, nNOS-IR was present in the both PMd and PMv, but neither ER-, nor ARO-IR were detected in the PM region. In the PMv of the intact female rat, no AR-IR was detected at 6 days of age, while it was detected as only a faint staining within 12 h after birth. When the male pups were castrated neonatally, no AR-IR was detected in the PMv. Subcutaneous injections of 5alpha-dihydrotestosterone (DHT) induced strong AR-IR in the castrated male and the intact female pups. On the contrary, the intensity of nNOS-IR stayed unchanged among these animals. Neonatal androgen and nitric oxide has been considered important to brain development. Moreover, involvement of the PMv in aggressive and mating behavior of male animals has been reported. Together with the fact that the AR-IR and nNOS-IR were found in the same neurons in the PMv, involvement of this nucleus in masculinization of the brain by non-aromatizable androgen is postulated.

Colocalization of neuronal nitric oxide synthase and androgen receptor immunoreactivity in the premammillary nucleus in rats.

To analyze the importance of premammillary region in male behavior such as aggression and mating, distribution of androgen- and estrogen-receptors (AR and ER), aromatase (ARO) and neuronal nitric oxide synthase (nNOS) was studied in the dorsal and ventral premammillary nuclei (PMd and PMv) of the rat by immunohistochemistry. The nNOS-immunoreactivities (-IR) were present both in the PMd and the PMv, while AR-IR were detected only in the PMv. AR-IR became undetectable after orchidectomy but they recovered by an injection of 5 alpha-dihydrotestosterone (DHT). In both nuclei, no clear signals of ER-IR were encountered. On the other hand, ARO-IR were found in the PMv but only very few. In the PMv, although DHT did not increase nNOS-IR significantly in castrated males, all the nNOS-IR cells contained AR-IR at least in intact and castrated-DHT injected males. Thus, involvement of nNOS-nitric oxide system in the PMv in the androgenic action on male behaviors was suggested.

Expression of estrogen receptor in the facial nucleus is suppressed by estradiol, but not by testosterone, indicating a lack of requirement for aromatization.

The transient expression of estrogen receptor (ER) in the ventromedial subnucleus of the facial nucleus was previously detected in the newborn rat, and the expression of ER molecules was down-regulated by daily injections of estradiol. Here we examined possible involvement of aromatization in this process. ER molecules were measured by immunohistochemistry and in situ hybridization histochemistry after daily injections of testosterone propionate (TP; 100 micrograms/0.02 ml) and estradiol benzoate (EB; 10 micrograms/0.02 ml) in the male pups castrated within 24 h of birth. Daily injections of TP for 5 consecutive days did not suppress ER and ER mRNA in the facial nucleus, while they were both suppressed by daily injections of EB. Moreover, aromatase immunoreactivity was not detected in the facial nucleus of both castrated, TP injected and intact control males at 6 days of age. The present findings therefore suggest that ER molecules expressed transiently in the facial nucleus are not directly involved in masculine sexual differentiation of the brain in newborn rat.

Flow cytometric analysis and immunohistochemistry of delayed-type hypersensitivity responses in mice immunized with rat hepatocytes.

Delayed-type hypersensitivity (DTH) responses to rat hepatocytes (HCs) in mice were investigated by flow cytometric analysis and immunohistochemistry with monoclonal antibodies directed against murine class II, CD4, and CD8 antigens. Mice were immunized subcutaneously (s.c.) with 10(6) rat HCs (referred to as s.c.-immunized mice), and control mice were injected s.c. with sterile Hanks' solution (non-immunized mice). Four days later, 10(5) rat HCs were injected into the footpad of s.c.-immunized mice and non-immunized mice. The DTH response in s.c.-immunized mice significantly increased after challenge when compared to that in non-immunized mice. The numbers of class II+, CD4+, and CD8+ cells in the footpad, and CD4+ and CD8+ cells in the inguinal lymph node of s.c.-immunized mice significantly increased during the DTH response. An increase in the number of CD4+ cells in the footpad of s.c.-immunized mice after challenge was more significant than that of non-immunized mice. The number of CD4+ cells increased more markedly in the footpad of s.c.-immunized mice as compared to that of CD8+ cells. Furthermore, immunohistochemical studies of the footpad of s.c.-immunized mice showed more severe infiltration of CD4+ cells rather than CD8+ cells at the injection site of rat HCs. These results suggest that the DTH response in the footpad of mice immunized with rat HCs is associated with severe infiltration of CD4+ cells.

Transient expression of estrogen receptor-immunoreactivity (ER-IR) in the layer V of the developing rat cerebral cortex.

Occurrence of estrogen receptor-immunoreactivity (ER-IR) in the cerebral cortex was examined in neonatal and adult rats. In newborn rats of postnatal day 1 (= day of birth) and postnatal day 5 (PD1 and PD5, respectively), ER-IR was not evident in the neocortex. On postnatal days 7, 10 and 13 (PD7, PD10 and PD13 respectively), a group of cells with distinct ER-IR appeared in the layer V of the auditory cortex. At the PD10, weak but specific ER-IR were also appeared in the somatosensory and the visual cortices. Among these areas, the ER-IR positive neurons occurred most frequently in the auditory cortex at PD10 rats. By examination of adjacent sections, one stained with Cresyl violet and the another stained with acethylcholinesterase (AChE) histochemistry, it was revealed that the region with ER-IR at PD7 to PD13 was limited to layer V of the neocortex. These signals, however, disappeared at PD15. In layer II of the neocortex, on the other hand, weak ER-IR signals were detected throughout the area sporadically at PD21 and in adults. The ER-IR detected transiently in the auditory cortex by the antiserum might contribute to maturation and establishment of the neurons of the rat auditory circuit.

Induction of substance P-immunoreactivity by estrogen in neurons containing estrogen receptors in the anterovental periventricular nucleus of female but not male rats.

Effects of gonadal steroids on numbers of neurons containing estrogen receptor (ER) and/or substance P (SP) were examined in the anteroventral periventricular nucleus (AVPV) of female and male rats by double-labeling immunohistochemistry employing antibodies specific for ER and SP. Animals were gonadectomized and received subcutaneously either oil alone (Control group), sequential injections of estradiol benzoate and oil (EB + Oil group), or those of EB and progesterone (EB + P group). In the female control rat, a large population of ER-immunoreactive (IR) cells were found clustered throughout the AVPV. They were counted more than 2,000 in total of 4 sections in this nucleus. On the contrary, SP-IR neurons were scarcely observed in the same area of this group. Administration of estrogen to female animals decreased the total number of ER-IR cells to 67% of the control group. In contrast to the supressive effect of estrogen to its own receptor, it induced SP-IR neurons in the AVPV of the female. Approximately 50-80 SP-IR neurons were counted in the 4 sections, and 59% of these neurons expressed ER-IR material in their nuclei. In the female EB + P group, the number of ER-IR neurons also decreased to 79% of the control group. Although the number of SP-IR neurons in this group decreased to 32% of that in the EB + Oil group, a ratio of coexistence of ER-IR material in these neurons increased to 75%. The male control group contained a smaller population of ER-IR cells relative to the female control (1497 vs 2143).(ABSTRACT TRUNCATED AT 250 WORDS)