Neural interaction of gonadotropin-regulating hormone immunoreactive neurons and the suprachiasmatic nucleus with the paraventricular organ in the Japanese grass lizard (Takydromus tachydromoides).

Our previous study demonstrated that the paraventricular organ (PVO) in the hypothalamus of the Japanese grass lizard (Takydromus tachydromoides) showed immunoreactivity against the light signal-transducing G-protein, transducin. This finding suggested that the PVO was a candidate for the deep-brain photoreceptor in this species. To understand functions of the PVO, we investigated distributions of transducin, serotonin, gonadotropin-releasing hormone (GnRH), and gonadotropin-inhibitory hormone (GnIH) in the lizard's brain. We immunohistochemically confirmed co-localization of transducin and serotonin in PVO neurons that showed structural characteristics of cerebrospinal fluid (CSF)-contacting neurons. GnRH-immunoreactive (ir) cells were localized in the posterior commissure and lateral hypothalamic area. Some of the serotonin-ir fibers extending from the PVO to the lateral hypothalamic area contacted the GnRH-ir cell bodies. GnIH-ir cells were localized in the nucleus accumbens, paraventricular nucleus, and upper medulla, and GnIH-ir fibers from the paraventricular nucleus contacted the lateral processes of serotonin-ir neurons in the PVO. In addition, we found that serotonin-ir fibers from the PVO extended to the suprachiasmatic nucleus (SCN), and the retrograde transport method confirmed the PVO projections to the SCN. These findings suggest that the PVO, by means of innervation mediated by serotonin, plays an important role in the regulation of pituitary function and the biological clock in the Japanese grass lizard.

Histological changes in immune and endocrine organs of quail embryos: exposure to estrogen and nonylphenol.

Effects of 17beta-estradiol and p-nonylphenol were examined in the immune and endocrine organs of Japanese quail embryos. The test compounds were injected into the yolk of embryonated eggs. Injection of estrogen resulted in (1) disappearance of lymphoid cells and flattened development of plicae in the bursa of Fabricius, (2) decreased area of thyroid follicles and height of simple cuboidal epithelial cells in the thyroid, (3) increased follicular appearance of the thymus, and (4) development of an ovotestis in male embryos. Injection of nonylphenol did not induce flattened plicae in the bursa of Fabricius or development of an ovotestis in male embryos, but it increased the disappearance of lymphoid cells from the lymphoid follicles in the bursa, decreased the height of simple cuboidal epithelial cells surrounding the thyroid follicle, and increased the follicle-like structure in the thymus in male embryos. These results suggest that nonylphenol has estrogenic effects, but these are low compared to those of estrogen itself.

Effects of estrogen (17 beta-estradiol) and p-nonylphenol on the development of immune organs in male Japanese quail.

The aim of this study was to determine the effects of estrogen and endocrine disrupters on the immune organs of Japanese quail. We used p-nonylphenol (NP) as a model endocrine disrupter. Birds were injected with 17 beta-estradiol (E2) in doses of 1, 0.1 and 0.01 ng/g body weight and NP in doses of 1000, 100 and 10 ng/g body weight from 4 to 7 weeks of age. The data indicate injection of E2 during the growth phase resulted in cyst formation, increased connective tissue, disappearance of lymphoid cells and disappearance of plicae in the bursa, disappearance of lymphocytes in the thymus and a greater population of trabecular arteries and veins in the spleen. Injection of NP tended to induce many empty vacuoles and increased connective tissue in the bursa of Fabricius but did not affect the structures of the spleen and thymus. We also studied whether apoptosis is involved in the histological changes of the bursa and found that apoptosis did not play an important role in the changes of the bursa. These results suggest that E2 and NP may affect the immune organs and the effect on the lymphoid tissue in immune organs is higher with E2 treatment than with NP.

Vitellogenin-immunohistochemistry in the liver and the testis of the medaka, Oryzias latipes, exposed to 17beta-estradiol and p-nonylphenol.

Vitellogenin (VTG) produced in male fish has been used for a biomarker to study endocrine disrupters. However, the characteristics of VTG produced in male fish have not been studied well. In this study, we investigated the localization of VTG in the liver and the testis of male medaka (Oryzias latipes) treated with 17beta-estradiol (E2) and p-nonylphenol (NP). The male fish were exposed to 1 microg/L E2 and 500 microg/L NP for 1-12 days. Control groups were kept in water including only vehicle. The frozen sections of the liver and the testis were stained with immunohistochemical methods using an antiserum against medaka VTG as the first antibody. In the E2 and NP treated liver, the hepatocytes showed immunoreactivity. In particular, the cytoplasm close to the cell membrane surrounding the sinusoids was strongly immunopositive. In the testis of both treatments, the interstitial tissues and the cells (spermatocytes) in the seminiferous tubules were immunopositive. The concentration of VTG became gradually higher in both tissues with longer treatments. These results suggest that germ cells in the testis treated with E2 and NP are able to incorporate and accumulate VTG.

Photoperiodic response of serotonin- and galanin-immunoreactive neurons of the paraventricular organ and infundibular nucleus in Japanese quail, Coturnix coturnix japonica.

We investigated the photoperiodic response of serotonin- and galanin (GA)- immunoreactive (ir) cells in the paraventricular organ (PVO) and infundibular nucleus (IF) of the Japanese quail and the interaction of these cells with gonadotropin-releasing hormone (GnRH)-ir neurons in the hypothalamus. Serotonin-ir cells were located in series from the PVO to the IF, and were connected with each other. The number of serotonin-ir cells differed significantly between light and dark phases on the short days (SD), but did not differ between light and dark phases on long days (LD). GA-ir cells were also found in the PVO and IF. The number of GA-ir cells under SD conditions was significantly greater than under LD conditions but did not change diurnally. Both serotonin-ir and GA-ir fibers ran along the GnRH-ir cells in the nucleus commissurae pallii. Serotonin-ir and GA-ir fibers were connected with the GnRH-ir fibers in the external layer of the median eminence (ME). We confirmed that GA-ir fibers were closely associated with serotonin-ir neurons in the PVO and IF. GA-ir neurons have at least 2 routes of regulating GnRH neurons directly, and indirectly via the serotonin-ir cells in the PVO and IF.