Trehalose prevents the formation of aggregates of mutant ataxin-3 and reduces soluble ataxin-3 protein levels in an SCA3 cell model.

Spinocerebellar ataxia type 3 (SCA3) is a neurodegenerative disorder caused by mutant ataxin-3 with an abnormally expanded polyQ tract and is the most common dominantly inherited ataxia worldwide. There are no suitable therapeutic options for this disease. Autophagy, a defense mechanism against the toxic effects of aggregation-prone misfolded proteins, has been shown to have beneficial effects on neurodegenerative diseases. Thus, trehalose, which is an autophagy inducer, may have beneficial effects on SCA3. In the present study, we examined the effects of trehalose on an SCA3 cell model. After trehalose treatment, aggregate formation, soluble ataxin-3 protein levels and cell viability were evaluated in HEK293T cells overexpressing ataxin-3-15Q or ataxin-3-77Q. We also explored the mechanism by which trehalose affects autophagy and stress pathways. A filter trap assay showed that trehalose decreased the number of aggregates formed by mutant ataxin-3 containing an expanded polyQ tract. Western blot and Cell Counting Kit-8 (CCK-8) results demonstrated that trehalose also reduced the ataxin-3 protein levels and was safe for ataxin-3-expressing cells, respectively. Western blot and total antioxidant capacity assays suggested that trehalose had great therapeutic potential for treating SCA3, likely through its antioxidant activity. Our data indicate that trehalose plays a neuroprotective role in SCA3 by inhibiting the aggregation and reducing the protein level of ataxin-3, which is also known to protect against oxidative stress. These findings provide a new insight into the possibility of treating SCA3 with trehalose and highlight the importance of inducing autophagy in SCA3.

Effects of music therapy on pain relief during fundus screening in infants: Randomized controlled clinical trial.

BACKGROUND: To determine the efficacy of music therapy on pain relief during fundus screening in infants. METHODS: The sample consisted of infants aged 0 to 3 months who required fundus screening. Infants were randomized to fast music, slow music, and control groups. All groups underwent fundus screening under topical anesthesia. Music therapy was provided to the music groups prior to, during, and after the operation. The patient's heart rate (HR), transcutaneous oxygen saturation, and crying decibel were measured. The Face, Legs, Activity, Cry, Consolability scale was used for pain measurement. RESULTS: A total of 300 subjects' data were collected. The quantitative analysis revealed that in both music groups, peripheral capillary oxygen saturation and satisfaction levels increased while pain scores decreased (P < .05). The slow music group's HR was shown to have significantly decreased (P < .05). CONCLUSION: Music therapy can effectively reduce pain and crying, and increase blood oxygen saturation during fundus examination of infants. Music with a rhythm of 60 to 80 beats per minute can decrease HR. Music therapy must be remembered to increase infants' comfort during fundus examination.

The cultivation regimes of Morchella sextelata trigger shifts in the community assemblage and ecological traits of soil bacteria.

The successful large-scale cultivation of morel mushrooms (Morchella sextelata) requires a comprehensive understanding of the soil bacterial communities associated with morel-farming beds, as the interactions between fungi and bacteria play a crucial role in shaping the soil microbiome. In this study, we investigated the temporal distribution and ecological characteristics of soil bacteria associated with morel fruiting bodies at different stages, specifically the conidial and primordial stages, under two cropping regimes, non-continuous cropping (NCC) and continuous cropping (CC). Our findings revealed a significant reduction in the yield of morel primordia during the third year following 2 years of CC (0.29 ± 0.25 primordia/grid), in comparison to the NCC regime (12.39 ± 6.09 primordia/grid). Furthermore, inoculation with morel mycelia had a notable impact on soil bacterial diversity, decreasing it in the NCC regime and increasing the number of generalist bacterial members in the CC regime. The latter regime also led to the accumulation of nutrients in the soil beds, resulting in a shift from a stochastic to a deterministic process in the composition of the bacterial community, which differed from the NCC regime. Additionally, mycelial inoculation had a positive effect on the abundance of potential copiotrophic/denitrifying and N-fixing bacteria while decreasing the abundance of oligotrophic/nitrifying bacteria. Interestingly, this effect was more pronounced in the NCC regime than in the CC regime. These results suggest that the increase in potential copiotrophic/denitrifying and N-fixing bacteria facilitated the decomposition of nutrients in exogenous nutrient bags by morel mushrooms, thereby maintaining nitrogen balance in the soil. Overall, our study provides valuable insights into the interactions between morel mycelia and the associated soil bacteriome as well as the influence of different cultivation regimes on these interactions. These findings contribute to our understanding of the complex dynamics of the soil microbiome and can inform strategies for optimizing morel mushroom cultivation.

The beneficial effects of Fomitopsis pinicola chloroform extract on a dextran sulfate sodium-induced ulcerative colitis mice model.

Background: As an intestinal non-specific inflammatory lesion, ulcerative colitis (UC) affects the health of many individuals. This study examined the possible beneficial effects of the chloroform extract of Fomitopsis pinicola (Swartz.: Fr) Karst (FPKc) on UC. Methods: The mice were given free access to drink with 4% dextran sulfate sodium (DSS) for 1 week to establish acute UC model. Next, 35 mg of FPKc or sulfasalazine (SASP) was given to the mice via gavage for 3 weeks. The disease activity index (DAI) and colonic mucosa damage index (CMDI) scores were calculated. The colon tissues of the mice were collected to measure the length and perform hematoxylin and eosin staining. The thymus and spleen indexes were determined. Interleukin (IL)-6, IL-8, tumor necrosis factor-α, aminotransferase (AST) and alanine aminotransferase (ALT) levels in the serum were determined. Results: FPKc or SASP treatment alleviated hematochezia and weight loss, ameliorated DAI and CMDI scores, and improved the crypt structure and length of the colon tissues. Relative to the UC model group, the spleen index in the FPKc group was reduced, which was accompanied by decreases of the IL-6 and IL-8 levels in the serum. FPKc also lowered the AST and ALT levels in the serum of the UC mice. Conclusions: FPKc protected the mice from DSS-induced UC injury. It may be that FPKc activates immune regulation and downregulates the expression of pro-inflammatory cytokines.

Clomiphene citrate treatment during perinatal development alters adult partner preference, mating behaviour and androgen receptor and vasopressin in the male mandarin vole Microtus mandarinus.

During development, many aspects of behaviour, including partner preferences and sexual behaviour, are "organized" by neural aromatization of androgen and oestrogen. This study aimed to analyse these processes in the mandarin vole (Microtus mandarinus); this is a novel mammalian model exhibiting strong monogamous pair bonds. Male pups were treated daily with a sesame oil only (MC) or the oestrogen receptor blocker-clomiphene citrate sesame oil mixture (MT) from prenatal day 14 to postnatal day 10. Female pups were treated daily with sesame oil only (FC). Partner preferences, sexual behaviour, and the expression of androgen receptor (AR) and arginine vasopressin (AVP) were examined when animals were 3 months old. The MT and FC groups exhibited male-directed partner preferences and feminized behaviour. AR-immunoreactive neurons (AR-IRs) in the medial preoptic area (mPOA), bed nucleus of stria terminalis (BNST), and medial amygdaloid nucleus (MeA) were reduced in MT males compared to MC males, and there was no significant difference in the number of AR-IRs between MT males and FC females. AVP-immunoreactive neurons (AVP-IRs) in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) were reduced in MT males compared to MC males, and there were no significant differences in the number of AVP-IRs between MT males and FC females. The results indicate a significant role of hormone signalling in the development of male mate preference in the novel monogamous mammal model.

Effects of treadmill exercise on anxiety-like behavior in association with changes in estrogen receptors ERα, ERß and oxytocin of C57BL/6J female mice.

Exercise can reduce the incidence of stress-related mental diseases, such as depression and anxiety. Control group was neither exposed to CVMS nor TRE (noCVMS/noTRE). Females were tested and levels of serum17-beta-oestradiol (E2), estrogen receptors α immunoreactive neurons (ERα-IRs), estrogen receptors ß immunoreactive neurons (ERß-IRs) and oxytocin immunoreactive neurons (OT-IRs) were measured. The results showed there's increased anxiety-like behaviors for mice from CVMS/noTRE, CVMS/higher speed TRE (CVMS/HTRE) and noCVMS/HTRE groups when they were put in open field and elevated maze tests. They had lower serum E2 levels than mice from CVMS/low-moderate speed TRE (CVMS/LMTRE), noCVMS/LMTRE and noCVMS/noTRE groups. The three groups of CVMS/noTRE, CVMS/HTRE and noCVMS/HTRE mice had more ERα-IRs and less ERß-IRs in the medial preoptic area (mPOA), bed nucleus of the stria terminalis (BNST) and medial amygdala (MeA), hypothalamic paraventricular nucleus (PVN) and supraoptic nucleus (SON). The number of OT-IRs in PVN and SON of CVMS/noTRE, CVMS/HTRE and noCVMS/HTRE mice was also lower than that of mice from CVMS/LMTRE, noCVMS/LMTRE and noCVMS/noTRE groups. Interestingly, CVMS/LMTRE and noCVMS/LMTRE mice were similar to noCVMS/noTRE mice in that they did not show anxiety, while CVMS/HTRE and noCVMS/HTRE mice did not, which were similar to the mice in CVMS/noTRE. We propose that LMTRE instead of HTRE changes the serum concentration of E2. ERß/ERα ratio and OT level in the brain may be responsible for the decrease in anxiety-like behavior in female mice exposed to anxiety-inducing stress conditions.

Charge-Transporting-Layer-Free, Vacuum-Free, All-Inorganic CsPbIBr2 Perovskite Solar Cells Via Dipoles-Adjusted Interface.

The inorganic perovskite has a better stability than the hybrid halide perovskite, and at the same time it has the potential to achieve an excellent photoelectric performance as the organic-inorganic hybrid halide perovskite. Thus, the pursuit of a low-cost and high-performance inorganic perovskite solar cell (PSC) is becoming the research hot point in the research field of perovskite devices. In setting out to build vacuum-free and carbon-based all-inorganic PSCs with the traits of simple fabrication and low cost, we propose the ones with a simplified vertical structure of FTO/CsPbIBr2/carbon upon interfacial modification with PEI species. In this structure, both the electron-transporting-layer and hole-transporting-layer are abandoned, and the noble metal is also replaced by the carbon paste. At the same time, FTO is modified by PEI, which brings dipoles to decrease the work function of FTO. Through our measurements, the carrier recombination has been partially suppressed, and the performance of champion PSCs has far exceeded the control devices without PEI modification, which yields a power conversion efficiency of 4.9% with an open circuit voltage of 0.9 V and a fill factor of 50.4%. Our work contributes significantly to give an available method to explore charge-transporting-layer-free, low-cost, and high-performance PSCs.

Transparent Ultrathin Metal Electrode with Microcavity Configuration for Highly Efficient TCO-Free Perovskite Solar Cells.

Optical microcavity configuration is one optical strategy to enhance light trapping in devices using planar electrodes. In this work, the potential application of optical microcavity configuration with ultrathin metal electrodes in highly efficient perovskite solar cells (PSCs) was investigated. By comparing with the device with conventional indium-tin-oxide (ITO) electrodes, it is shown that by carefully designing the Ag/dielectric planar electrode, a device with an optical microcavity structure can achieve comparable-or even higher-power conversion efficiency than a conventional device. Moreover, there is a relative high tolerance for the Ag film thickness in the optical microcavity structure. When the thickness of the Ag film is increased from 8 to 12 nm, the device still can attain the performance level of a conventional device. This gives a process tolerance to fabricate devices with an optical microcavity structure and reduces process difficulty. This work indicates the great application potential of optical microcavities with ultrathin metal electrodes in PSCs; more research attention should be paid in this field.

Divalproex sodium regulates ataxin-3 translocation likely by an importin α1-dependent pathway.

Nuclear localization of ataxin-3 plays a fundamental role in seeding aggregation and the pathology of spinocerebellar ataxia type 3 (SCA3). However, very few compounds that are able to modulate the nuclear transport of ataxin-3 have been identified. In our previous study, we found that divalproex sodium (DVS) reduced heat shock-induced nuclear localization of ataxin-3. However, the mechanism of DVS in the translocation of ataxin-3 still remains unknown. There is accumulating evidence that importins are regulated by acetylation, and histone deacetylase inhibitors can interrupt this process. With this in mind, we used cells coexpressing ataxin-3 and importin α1 (encoded by KNPA2) to probe whether ataxin-3 is the shuttling cargo of importins and whether DVS plays a role in the nuclear transport of ataxin-3 through the transport protein pathway. Here, we reported that importin α1 enhanced nuclear amount of ataxin-3 and increased the aggregate formation and that DVS restored it to the normal level. Importantly, ataxin-3 is shown to directly bind to importin α1. Moreover, DVS modulated the function of importin α1 likely by altering its localization. We believe that this study provides a proof of principle for addressing the mechanism of DVS and furthers our understanding of the role of importins in the nuclear accumulation of ataxin-3 in SCA3.

Estradiol Treatment during Perinatal Development Alters Adult Partner Preference, Mating Behavior and Estrogen Receptors α and ß in the Female Mandarin Vole (Microtus mandarinus).

During development, many aspects of behavior, including partner preferences and sexual conduct, are "organized" by estradiol. This study aimed at analyze these processes in the mandarin vole (Microtus mandarinus), a novel experimental mammal with strong monogamous pair bonds. Female pups were treated daily with an oil vehicle (FC) or ß-Estradiol (E2, FT) from prenatal day 14 to postnatal day 10. Male pups were treated daily with the oil vehicle only (MC). Partner preferences, sexual conduct and the expression of estrogen receptors α (ERα) and ß (ERß) were examined when animals were 3 months old. FT and MC groups showed female-directed partner preferences and masculinized behavior. ERα- immunoreactive neurons (ERα-IRs) in the bed nucleus of stria terminalis (BNST) and medial amygdaloid nucleus (MeA) was greater in FT females than MC males, and there was no significant difference in the number of ERα-IRs between FT and FC females. No difference was found for ERα-IRs in the preoptic area (mPOA) or ventromedial nucleus of the hypothalamus (VMH) of FT females or MC males, and they were significantly fewer than in FC females. ERß-immunoreactive neurons (ERß-IRs) in these four brain regions did not alter the ERß/ERα ratio in different brain regions during perintal developments. However, the number of ERß-IRs in FT females and MC males were greater than in FC females. We propose that estradiol treatment during perinatal development is responsible for adult partner preferences and mating behavior.

Postnatal separation prevents the development of prenatal stress-induced anxiety in association with changes in oestrogen receptor and oxytocin immunoreactivity in female mandarin vole (Microtus mandarinus) offspring.

Oestrogen has both anxiogenic and anxiolytic effects because of variation in opposing action on alpha (ERα) and beta (ERß) estrogen receptors in the medial preoptic area (mPOA), bed nucleus of the stria terminalis (BNST) and medial amygdala (MeA). Oxytocin (OT) reverses some of the anxiogenic effects of oestrogen in the hypothalamic paraventricular nucleus (PVN) and supraoptic nucleus (SON). Because anxiety disorders are twice as common in women as in men, and oestrogen and OT are more important in females, we examined interactions between prenatal restraint stress (GS) and postnatal early short-term maternal separation (MS) and female mandarin vole behaviour, estrogen receptors and OT. The results show that adult female offspring from GS/noMS mothers showed increased anxiety in open-field and elevated plus-maze tests and had lower serum 17-beta-oestradiol (E2 ) levels than female offspring from GS/MS, noGS/MS and noGS/noMS mothers. GS/noMS females had more immunoreactive neurons for ERα in several brain regions and less ERß- and OT-immunoreactive neurons in brain areas compared to GS/MS, noGS/MS and noGS/noMS offspring. Interestingly, noGS/MS and GS/MS offspring were similar to noGS/noMS offspring in that they did not develop anxiety as adults. We propose that MS alters the serum concentration of E2 and that the ERß/ERα ratio and OT level in the brain may be responsible for the decrease in anxiety-like behaviour in adult female offspring initially exposed to anxiety-inducing conditions via an adverse foetal environment.

Early bi-parental separation or neonatal paternal deprivation in mandarin voles reduces adult offspring paternal behavior and alters serum corticosterone levels and neurochemistry.

Although the effect of early social environments on maternal care in adulthood has been examined in detail, few studies have addressed the long-term effect on paternal care and its underlying neuroendocrine mechanisms. Here, using monogamous mandarin voles (Microtus mandarinus) that show high levels of paternal care, the effects of early bi-parental separation (EBPS) or neonatal paternal deprivation (NPD) on adult paternal behavior, serum corticosterone levels, and receptor mRNA expression in the nucleus accumbens (NAcc) and medial preoptic area (MPOA) were investigated. Compared to the parental care group (PC), we found that EBPS reduced crouching behavior and increased inactivity, self-grooming, and serum corticosterone levels in adult offspring; and NPD significantly reduced retrieval behavior and increased self-grooming behavior of offspring at adulthood. EBPS displayed more dopamine type I receptor (D1R) mRNA expression in the NAcc, but less oxytocin receptor (OTR) mRNA expression than PC in the MPOA. Both EBPS and NPD exhibited more mRNA expression of estrogen receptor alpha (ERα) than PC in the MPOA. In the EBPS group, increased serum corticosterone concentration was closely associated with reduced crouching behavior, and reduced expression of OTR was closely associated with altered crouching behavior and increased D1R expression. Our results provide substantial evidence that EBPS or NPD has long-term consequences and reduces paternal behavior in adult animals. Importantly the oxytocin system in the MPOA might interact with NAcc dopamine systems to regulate paternal behavior and EBPS may affect interactions between the MPOA and NAcc.

Early paternal deprivation alters levels of hippocampal brain-derived neurotrophic factor and glucocorticoid receptor and serum corticosterone and adrenocorticotropin in a sex-specific way in socially monogamous mandarin voles.

In monogamous mammals, fathers play an important role in the development of the brain and typical behavior in offspring, but the exact nature of this process is not well understood. In particular, little research has addressed whether the presence or absence of paternal care alters levels of hippocampal glucocorticoid receptor (GR) and brain-derived neurotrophic factor (BDNF), and basal levels of serum corticosterone (CORT) and adrenocorticotropin (ACTH). Here, we explored this concept using socially monogamous mandarin voles (Microtus mandarinus), a species in which fathers display high levels of paternal care toward their pups. Our immunohistochemical study shows that paternal deprivation (PD) significantly decreased levels of GR and BDNF protein in the CA1 and CA2/3 of the hippocampus. In the dental gyrus, decreases in GR and BDNF induced by PD were evident in females but not in males. Additionally, enzyme-linked immunosorbent assay results show that PD significantly upregulated levels of serum CORT and ACTH in females, but not males. These findings demonstrate that PD alters HPA axis activity in a sex-specific way. The changes in stress hormones documented here may be associated with alteration in hippocampal BDNF and GR levels.

Study of Fos, androgen receptor and testosterone expression in the sub-regions of medial amygdala, bed nucleus of stria terminalis and medial preoptic area in male Mandarin voles in response to chemosensory stimulation.

In many rodent species, including mandarin voles (Microtus mandarinus), the behavioral response to odors is regulated by a network of steroid-sensitive ventral forebrain nuclei including the medial amygdala (Me), bed nucleus of the striaterminalis (BNST), and medial preoptic area (MPOA). Although it is well-known that Me, BNST, and MPOA are closely interconnected, function independently in regulating odor-guided social behaviors, little is known about how order information is processed in the sub-regions of Me, BNST, and MPOA. In order to answer this question, we let male mandarin voles expose to two different odors including female vaginal fluid (FVF) and male flank gland secretion (MFGS) and detect the expression of Fos, androgen receptor (AR) and testosterone (T) in the sub-regions of Me, BNST, and MPOA. We found that FVF stimulus caused increased Fos, AR and T expression in the posterior subdivision of the Me (MeP), the posterior medial subdivision of the BNST (BNSTpm), and the medial preoptic nucleus (MPN), while MFGS stimulus did not change Fos, AR and T expression neither in the MeP, BNSTpm, and MPN nor in the anterior subdivision of the Me (MeA), the posterointermediate subdivision of the BNST (BNSTpi), and the lateral subdivision of the MPOA (MPOAl). Serum testosterone levels were increased after 1h in males exposed to FVF. This study provides insight in understanding the relationship between female odor stimulation and Fos, AR and T expression in specific brain areas in males, and the regulatory role of testosterone in this biochemical process.

[Perinatal clomiphene citrate treatment changes sexual orientations of male mice].

Perinatal period and adolescence are critical for brain development, which is the biological basis of an individual's sexual orientation and sexual behavior. In this study, animals were divided into two groups and their sexual orientations were observed: one group experienced drug treatments during the perinatal period, and the other group was castrated at puberty. The results showed that estradiol treatment had no effect on mature male offspring's sexual orientations, but 9 days and 14 days of clomiphene citrate treatment significantly increased the chance of homosexuality and effeminized behavior. In addition, the sexual orientation of mature normal male offspring, which were castrated when they were 21 days old,was not significant different from the control animals. These findings suggest that the inhibition of perinatal estrogen activities could suppress individual male-typical responses, enhance female-typical responses and induce homosexual orientations. Moreover, the masculinizing effects of estrogen were more obvious during perinatal period than adolescence.

Relationship between sexual satiety and motivation, brain androgen receptors and testosterone in male mandarin voles.

Androgen receptors participate in the neuroendocrine regulation of male sexual behavior, primarily in brain areas located in the limbic system. Males of many species present a long-term inhibition of sexual behavior after several ejaculations, known as sexual satiety. It has been shown in rats that androgen receptor expression is reduced 24h after a single ejaculation, or mating to satiety, in the medial preoptic area, nucleus accumbens and ventromedial hypothalamus. The aim of this study was to analyze these processes in another animal, the mandarin vole (Microtus mandarinus). We compared differences in androgen receptor (AR) and testosterone (T) expression in various brain areas between male mandarin voles sexually satiated and those exposed to receptive females but not allowed to mate. Sexual satiety was associated with decreased AR and T expression in the lateral septal nucleus (LS), medial amygdala (MeA), medial preoptic area (mPOA) and ventromedial hypothalamic nucleus (VMH). Males exposed to receptive females showed an increase in AR and T expression in the bed nucleus of the stria terminalis (BNST), LS, MeA and VMH. Serum testosterone levels remained unchanged after 24h in males exposed to receptive females or males mated to satiety. These data suggest a relationship between sexual activity and a decrease in AR and T expression in specific brain areas, and a relationship between sexual motivation and increased AR and T expression in other brain areas, independently of testosterone levels.

The effects of neonatal paternal deprivation on pair bonding, NAcc dopamine receptor mRNA expression and serum corticosterone in mandarin voles.

High levels of paternal care are important for the development of social behavior in monogamous rodents. However, the effects of paternal care on the formation of pair bonding and underlying neuroendocrine mechanisms, especially the involvements of dopamine system and corticosterone, are not well understood. We investigated effects of paternal deprivation on pair bonding in mandarin voles (Microtus mandarinus), a socially monogamous rodent. Paternal deprivation was found to inhibit the formation of pair bonding in females according to partner preference tests (PPT). Paternal deprivation also reduced body contact behavior and increased aggression in males and females in PPT. During social interaction tests (SIT), paternal deprivation was found to reduce investigative and aggressive behaviors but increase body contact and self-grooming in females, and reduce staring, aggression, body contact and self-grooming in males when interacting with the opposite sex. Paternal deprivation reduced the expression of dopamine 1-type receptor (D1R) mRNA and dopamine 2-type receptor (D2R) mRNA in the nucleus accumbens of female offspring in later life, but enhanced mRNA expression of these two dopamine receptors in males. After three days of cohabitation the expression of D1R mRNA and D2R mRNA was negatively correlated for voles reared by two parents, but positively correlated in paternally deprived animals. Paternal deprivation reduced serum corticosterone levels in females but had the opposite effect in males. Three days of cohabitation did not alter corticosterone levels of PD females, but reduced it in PC females. Our results provide substantial evidence that paternal deprivation inhibits the formation of pair bonding in female mandarin voles and alters social behavior later in life. These behavioral variations were possibly associated with sex-specific alterations in the expression of two types of dopamine receptors and serum corticosterone levels induced by paternal deprivation.

Effects of castration on aggression and levels of serum sex hormones and their central receptors in mandarin voles (Microtus mandarinus).

Aggression in socially monogamous mandarin vole (Microtus mandarinus) was observed after castration. Levels of serum sex hormones and their central receptors were also measured using enzyme-linked immunosorbent assay and immunohistochemistry methods. The data indicate that adult males showed higher levels of aggression after castration. However, castration significantly reduced levels of serum testosterone, and the number of androgen receptor immunoreactive neurons in the anterior hypothalamus, bed nucleus of the stria terminalis, medial amygdaloid nucleus (P<0.01) and lateral septal nucleus (P<0.05). In addition, levels of estrogen receptor ß in the anterior hypothalamus and medial amygdaloid nucleus (P<0.05), bed nucleus of the stria terminalis and lateral septal nucleus (P<0.01) declined to varying degrees at weekly intervals. In contrast, serum 17ß-estradiol concentrations were up-regulated by castration and castration did not change levels of estrogen receptor α in the medial amygdaloid nucleus and lateral septal nucleus, but increased it in the anterior hypothalamus 3 weeks after castration (P<0.05). We suggest that higher levels of aggression induced by castration may be independent of serum testosterone and androgen receptors, and may be associated with high serum 17ß-estradiol concentrations, stable estrogen receptor α immunoreactive neurons in some brain regions and the relative ratio of the two estrogen receptors.