Social defeat stress-specific increase in c-Fos expression in the extended amygdala in mice: Involvement of dopamine D1 receptor in the medial prefrontal cortex.

We recently reported that dopamine D1 receptor in the medial prefrontal cortex (mPFC) is activated by subthreshold social defeat stress and suppresses the induction of depressive-like behavior in mice. However, which mPFC projection(s) mediates this antidepressant-like effect remains poorly understood. Here we show that social defeat stress specifically increased c-Fos expression, a marker for neuronal activity, in distinct brain regions involved in emotional regulation, relative to novelty-induced exploration. Among these brain areas, D1 knockdown in the mPFC decreased social defeat stress-induced c-Fos expression in the interstitial nucleus of the posterior limb of the anterior commissure (IPAC), a subregion of the extended amygdala. Using retrograde adeno-associated virus vectors and transgenic mice expressing Cre recombinase under the D1 promoter, we also found that D1-expressing deep-layer pyramidal neurons in the mPFC send direct projections to the IPAC. These findings indicate that social defeat stress specifically activates neurons in distinct brain areas, among which the IPAC is regulated by dopamine D1 receptor in the mPFC perhaps through direct projections. Thus, this study provides hints toward identifying neural circuits that underlie antidepressant-like effects of stress-induced dopamine D1 receptor signaling in the mPFC.

A role for mDia, a Rho-regulated actin nucleator, in tangential migration of interneuron precursors.

In brain development, distinct types of migration, radial migration and tangential migration, are shown by excitatory and inhibitory neurons, respectively. Whether these two types of migration operate by similar cellular mechanisms remains unclear. We examined neuronal migration in mice deficient in mDia1 (also known as Diap1) and mDia3 (also known as Diap2), which encode the Rho-regulated actin nucleators mammalian diaphanous homolog 1 (mDia1) and mDia3. mDia deficiency impaired tangential migration of cortical and olfactory inhibitory interneurons, whereas radial migration and consequent layer formation of cortical excitatory neurons were unaffected. mDia-deficient neuroblasts exhibited reduced separation of the centrosome from the nucleus and retarded nuclear translocation. Concomitantly, anterograde F-actin movement and F-actin condensation at the rear, which occur during centrosomal and nuclear movement of wild-type cells, respectively, were impaired in mDia-deficient neuroblasts. Blockade of Rho-associated protein kinase (ROCK), which regulates myosin II, also impaired nuclear translocation. These results suggest that Rho signaling via mDia and ROCK critically regulates nuclear translocation through F-actin dynamics in tangential migration, whereas this mechanism is dispensable in radial migration.

Deficiency of mDia, an actin nucleator, disrupts integrity of neuroepithelium and causes periventricular dysplasia.

During development of the central nervous system, the apical-basal polarity of neuroepithelial cells is critical for homeostasis of proliferation and differentiation of neural stem cells. While adherens junctions at the apical surface of neuroepithelial cells are important for maintaining the polarity, the molecular mechanism regulating integrity of these adherens junctions remains largely unknown. Given the importance of actin cytoskeleton in adherens junctions, we have analyzed the role of mDia, an actin nucleator and a Rho effector, in the integrity of the apical adherens junction. Here we show that mDia1 and mDia3 are expressed in the developing brain, and that mDia3 is concentrated in the apical surface of neuroepithelium. Mice deficient in both mDia1 and mDia3 develop periventricular dysplastic mass widespread throughout the developing brain, where neuroepithelial cell polarity is impaired with attenuated apical actin belts and loss of apical adherens junctions. In addition, electron microscopic analysis revealed abnormal shrinkage and apical membrane bulging of neuroepithelial cells in the remaining areas. Furthermore, perturbation of Rho, but not that of ROCK, causes loss of the apical actin belt and adherens junctions similarly to mDia-deficient mice. These results suggest that actin cytoskeleton regulated by Rho-mDia pathway is critical for the integrity of the adherens junctions and the polarity of neuroepithelial cells, and that loss of this signaling induces aberrant, ectopic proliferation and differentiation of neural stem cells.

Determination and characterization of hydroxylated polychlorinated biphenyls (OH-PCBs) in serum and adipose tissue of Japanese women diagnosed with breast cancer.

In the present study, we analyze and characterize the levels of PCBs and OH-PCBs in serum and breast adipose tissue, and measure TH (total 3,5,5'-triiodothyronine (tT3) and total l-thyroxine (tT4)) levels in serum of Japanese women diagnosed with breast cancer (n = 51) to know how PCBs affect thyroid hormone homeostasis. Concentrations of Sigma(90)OH-PCBs in serum and adipose tissues were 630 pg g(-1) serum wet wt. and 140 pg/ g(-1) lipid wt., respectively, and the levels of OH-PCBs in adipose tissues were 4-5 times lower than in serum. The tT3 and tT4 concentrations in serum (n = 20) were 1.3 ng g(-1) serum wet wt. and 62 ng g(-1) serum wet wt., respectively. Among all the OH-PCBs, 4'-OH-CB101/120, 4-OH-CB107/4'-OH-CB108, 3'-OH-CB138, 4-OH-CB146, 3-OH-CB153, 4'-OH-CB172, and 4-OH-CB187 were the dominant isomers in serum. No significant correlation between the concentrations of the TH and OH-PCBs in serum was observed (p > 0.05). The results indicated that the relationship between the OH-PCB levels and the thyroid hormone markers was rare. From analysis of two kinds of human biological samples, it was estimated that OH-PCBs have little effect on thyroid hormone homeostasis.

Hydroxylated polychlorinated biphenyls (OH-PCBs) induce vitellogenin through estrogenic activity in primary-cultured hepatocytes of the Xenopus laevis.

Vitellogenin (VTG)-inducing activities of 21 kinds of hydroxylated polychlorinated biphenyls (OH-PCBs) were investigated by an assay system using the primary-cultured hepatocyte of the adult male Xenopus laevis. An enzyme-linked immunoabsorbent assay (ELISA) was able to detect VTG at a minimum detection limit of 0.2ngmL(-1). The levels of VTG production in hepatocytes of male X. laevis exposed to six kinds of OH-PCB isomers (2'-OH-CB30, 3'-OH-CB30, 4'-OH-CB30, 4'-OH-CB50, 4'-OH-CB61 4'-OH-CB65) were significantly higher as compared to the control group (p<0.05). The relative ranking of the OH-PCBs was E2 (100%), 4'-OH-CB30 (2.0%)>2'-OH-CB30 (0.15%)>4'-OH-CB61 (0.046%)>3'-OH-CB30 (0.013%)>4'-OH-CB50 (0.002%)>4'-OH-CB65 (0.0001%). Although there were some differences between the estrogenicity values by the VTG-inducing activities and yeast two-hybrid assay for human estrogen receptor alpha (ERalpha). The results from VTG-assays suggested that an important factor for determining estrogenicity is the positions of the hydroxyl group and chlorine with the highest activity resulting from a para-hydroxyl group. The OH-PCB structures of high rank order in the VTG-assay had no chlorine substituted phenolic ring. Moreover, the meta-chlorine substituted phenolic ring is an important factor in determining estrogenicity. The OH-PCBs with meta-chlorine substituted phenolic ring did not show estrogenicity during VTG-assay.

In vitro evaluation of atmospheric particulate matter and sedimentation particles using yeast bioassay system.

Little information on the evaluation of airborne particulate matter (APM) and sedimentation particles from subway stations is available. The thermal metamorphism of train wheels generating toxic particles in subway stations is a possibility. In this study, the toxicity and physiological effects of particles from subway stations were evaluated using a yeast bioassay system. Estrogenic and antiestrogenic activities of APM in APM extracts from subway stations were determined. No estrogenic activity was found in the APM fractions and their S9-activated APM samples. Sedimentation dust samples also showed no estrogen activity. In contrast, extracts from sedimentation dust samples showed antiestrogen activity. Marked yeast toxicity was observed in the samples extracted from sedimentation dust. Potent yeast toxicity was also found in the S9-activated extracts from sedimentation dust. The results suggest that sedimentation dust from a semiclosed area of a subway system has antiestrogen activity, although both the origin and generation system of this activity are uncertain. These pollutants in sedimentation dust may change to a more toxic form in vivo by S9 activation.

Endocrine effects generated by photooxidation of coplanar biphenyls in water using titanium dioxide.

The oxidative photodegradation behaviors of selected three coplanar polychlorinated biphenyls (PCBs), (CB77, CB81, and CB169) using titanium dioxide (TiO(2)) in water were investigated. The main purposes were to clarify the structural relation between the original PCBs and the intermediates derived by TiO(2) oxidation and to evaluate the estrogenic and thyroid hormonal activity in the treated three coplanar PCBs during the oxidative reactions. Approximately 90% of the three coplanar PCBs decomposed within 180 min. Intermediates from the decomposition of the three coplanar PCBs, such as some hydroxylated-PCBs (OH-PCBs), carboxylic intermediates, phenolic intermediates, and other intermediates produced by the cleavage of a benzene ring were identified and quantified. In the degradation pathways, the produced amounts of OH-PCBs increased within 60 min of irradiation time. The estrogenic activity and thyroid hormonal activity of the intermediates from the three coplanar PCBs in water at 0, 60, 120, 180 and 240 min of irradiation time were assessed by using a yeast two-hybrid assay system for human estrogen receptor alpha (hERalpha) and human thyroid hormone receptor alpha (hTRalpha). The maximal estrogenic activities were induced by the solutions of decomposed PCBs with irradiation time at 60-120 min similar and slightly lower than those after the irradiation time. We found that the solutions occuring during the irradiation times of 60-120 min contained several 4-OH-PCBs substituted with OH and Cl at para- and para'-positions having estrogenic activity. The thyroid hormonal activity was not detected in the decomposed three coplanar PCBs solutions.

Oxidative degradation of BPA using TiO2 in water, and transition of estrogenic activity in the degradation pathways.

The oxidative degradation behavior of bisphenol A (BPA) using titanium dioxide (TiO(2)) in water was investigated. The main purposes were to clarify the relationship with estrogenic activity from the degradation pathways demonstrated by oxidation of BPA in water. Approximately 99% of the BPA decomposed within 300 min, and gas chromatography (GC) mass spectrometry (MS) and liquid chromatography (LC) MS analysis revealed many intermediates during the degradation process. Intermediates by decomposition of BPA, such as hydroxylated-BPA (OH-BPA), carboxylic intermediates, phenolic intermediates, and other intermediates produced by the cleavage of a benzene ring were identified and quantified. Estrogenic activities of the degradation pathways of the BPA in water were assessed by using a constructed yeast two-hybrid assay system for human estrogen receptor alpha (hERalpha) and Japanese medaka fish (Oryzias latipes) estrogen receptor alpha (medERalpha). Estrogenic activity for hERalpha and medERalpha was reduced to less than 20% of the initial activity for BPA after 240 min of UV irradiation. However, estrogenic activity for medERalpha was increased by 110% from the initial activity for BPA at 60 min of UV irradiation. It was estimated that medERalpha assay was more sensitive for BPA and the intermediates than was the hERalpha assay. From these findings, we estimate that the intermediates by the oxidation of BPA have the behaviors of xenoestrogen to the aquatic wildlife in the environment.

Production mechanism of hydroxylated PCBs by oxidative degradation of selected PCBs using TiO2 in water and estrogenic activity of their intermediates.

The oxidative photodegradation behaviors of selected polychlorinated biphenyls (PCBs) [2,3,3',4'-tetraCB (BZ number: CB56), 2,3',4',5-tetraCB (CB70), and 3,4,4',5-tetraCB (CB81: coplanar PCB)] using titanium dioxide (TiO2) in water were investigated. The main purposes were to clarify the structural relation between the original PCBs and the intermediates derived by TiO2 oxidation and to evaluate the estrogenic activity in the treated PCBs during the oxidative reactions. Approximately 90% of the three tetraCBs decomposed within 120 min. Intermediates by decomposition of three tetraCBs, such as some OH-tetraCBs and OH-triCBs, carboxylic intermediates, phenolic intermediates, and other intermediates produced by the cleavage of a benzene ring were identified and quantified. In the degradation pathways, the produced amounts of OH-tetraCB and OH-triCB increased within 60 min of irradiation time. Estrogenic activities of the intermediates from the three tetraCBs in water were assessed by using a yeast two-hybrid assay system for human estrogen receptor alpha (hERalpha). The maximal estrogenic activities were induced by the solutions of decomposed CB81 with irradiation time at 60 min. We found that the solutions at an irradiation time of 60-120 min contained several 4-OH-tetraCBs and 4-OH-triCBs substituted with OH and Cl at para- and para'-positions. It is presumed that the chemical structures of the 4-OH-PCBs are similar to that of 17beta-estradiol (beta-E2); these intermediates present strong estrogenic activities. Moreover, we learned that there is a high possibility of conversion from some low toxic PCBs congeners to strong estrogenic OH-PCBs.

A rapid and sensitive analysis of diphenylarsinic acid in water by gas chromatography/mass spectrometry.

A rapid and sensitive analytical method using pentafluorothiophenol (PFTP) derivatization was applied to detect diphenylarsinic acid (DPAA) in water. In this study, the optimum derivatization conditions, such as acid concentration, reaction time and reaction temperature, were investigated to develop a suitable procedure for DPAA determination. After extracting the derivatives into benzene, the determination was carried out by gas chromatography/mass spectrometry (GC/MS) with selected ion monitoring (SIM). The detection limit of the method was 9.4 microg/l, and the overall recoveries obtained from real environmental samples were 88.9 - 104.7% and coefficient variations were 5.1 - 13.9%.

Effect of estrogenic activity, and phytoestrogen and organochlorine pesticide contents in an experimental fish diet on reproduction and hepatic vitellogenin production in medaka (Oryzias latipes).

Endocrine-disrupting chemicals (EDCs) are giving rise to serious concerns for humans and wildlife. Phytoestrogens, such as daidzein and genistein in plants, and organochlorine pesticides are suspected EDCs, because their chemical structure is similar to that of natural or synthetic estrogens and they have estrogenic activity in vitro and in vivo. We assessed estrogenic activity and dietary phytoestrogen and organochlorine pesticide contents of various fish diets made in the United Kingdom, and compared them with those features of diets made in Japan that were tested in a previous study. Genistein and daidzein were detected in all of the diets. Using an in vitro bioassay, many of these diets had higher activation of estrogen beta-receptors than estrogen alpha-receptors. Organochlorine pesticides such as hexachlorobenzene, beta-benzene hexachloride (BHC), and gamma-BHC were detected in all fish diets. On the basis of these data, we investigated the effect of differing dietary phytoestrogen content in Japanese fish diets on hepatic vitellogenin production and reproduction (fecundity and fertility) in medaka (Oryzias latipes). Assessment of the effects of a 28-day feeding period on reproduction of paired medaka did not indicate significant differences in the number of eggs produced and fertility among all feeding groups. However, hepatic vitellogenin values were significantly higher for male medaka fed diet C (genistein, 58.5 +/- 0.6 microg/g; daidzein, 37.3 +/- 0.2 microg/g) for 28 days compared with those fed diet A (genistein, < 0.8 microg/g; daidzein, < 0.8 microg/g) or diet B (genistein, 1.4 +/- 0.1 microg/g; daidzein, 2.0 +/- 0.1 microg/g). Our findings indicate that fish diets containing high amounts of phytoestrogens, such as diet C, have the potential to induce hepatic vitellogenin production in male medaka, even if reproductive parameters are unaffected. Therefore, some diets, by affecting vitellogenin production in males, may alter estrogenic activity of in vivo tests designed to determine activity of test compounds added to the diet.