Molecular mechanisms of sex determination and gonadal sex differentiation in fish

We have used various genetic and molecular approaches to investigate the mechanisms of sex determination and gonadal sex differentiation in fish. DMY was identified as the sex-determining gene of medaka. In tilapia, endogenous estrogens act as natural inducers of ovarian differentiation, while DMRT1 may be important for testicular differentiation. The roles of these regulators in sex determination and gonadal sex differentiation were ascertained using a gene or hormonal blockade strategy.     

Serological properties and intrinsic antibiotic resistance of soybean bradyrhizobia isolated from Thailand

A group of Bradyrhizobium strains isolated from soybean plants in Thailand did not correspond to any known DNA homology groups of Bradyrhizobium japonicum and Bradyrhizobium elkanii reported by Hollis et al. (J. Gen. Microbiol., 123, 215–222, 1981). To clarify the phenotypic characteristics of the group, serological properties and intrinsic antibiotic resistance (IAR) profile of 94 Thai strains were compared with those of USDA and Japanese strains. Indirect ELISA tests for each Thai strain were performed agaiIl.st polyclonal antisera prepared against 15 USDA standard serotype strains of B. japonicum and B. elkanii. Among the 94 Thai strains tested, 36 which were previously identified as B. elkanii, with the exception of one strain, were strongly responsive to an antiserum prepared against USDA 31. The remaining 58 strains, with the exception of two strains, showed multiple cross reactions which were peculiar to the Thai strains. These serological reaction patterns did not correspond to any known serogroups labeled as B. japonicum and B. elkanii. In the IAR test, the taxonomically unknown Thai soybean bradyrhizobia exhibited a high level of resistance to neomycin (50 µg/mL), polymyxin (50 µg/mL), nalidixic acid (15 µg/mL), and kanamycin (15 µg/mL). Kanamycin could thus be useful in combination with neomycine and nalidixic acid for distinguishing between the unknown Thai strains and strains of B. japonicum and B. elkanii. Our results demonstrated that the unknown Thai strains were serologically and IAR-phenotypically remote from both B. japonicum and B. elkanii.     

The comparison of the collagen fiber contents and hepatic stellate cell distribution in male and female chicken livers

The difference in collagen fiber content, morphological properties and hepatic stellate cell (HSC) distribution was investigated in the liver of both sexes in chicken. Collagen fiber specimens were obtained by maceration treatment with NaOH solution. HSCs were detected using desmin‐specific immunohistochemistry. The ratio of liver weight to body weight was larger in the female than the male chickens. Collagen fiber content, the numerical density of HSCs and the percentage area displaying desmin immunopositivity were not different between the right and left lobes of the liver, in both male and female chickens. However, all of these parameters were larger in the males than the females. In the light microscopic observation, many HSCs in the male had large and elongated cytoplasmic processes. Conversely, HSCs with poorly developed cytoplasmic processes were frequently observed in females. Liver tissue is structurally stronger in male chickens than females and the activity and density of HSCs may be related to the collagen fiber content in chicken liver.     

Proteome analysis: a new approach to identify key proteins involved in acquisition of maturational competence and oocyte maturation of medaka oocytes

We have applied analytical methods of proteomics to identify key proteins that are involved in physiologically important processes on fish reproduction. We have improved methods of proteome analysis for fish oocytes and revealed changes of expression patterns of oocyte proteins and protein phosphorylation during acquisition of maturational competence and signal transduction of oocyte maturation of medaka oocytes.     

Food habits of sika deer as assessed by fecal analysis in Mt. Ohdaigahara,central Japan

Food habits of sika deer,Cervus nippon centralis temminck, in Mt. Ohdaigahara, central Japan, were investigated by the fecal analysis method from September, 1990, to June, 1991. The fecal analysis showed that the percentage ofSasa nipponica Makino et Shibata in the fecal composition was approximately 50% throughout the year, indicating that it was the main food plant. Bark and twigs were also found in the feces in all seasons, suggesting that the deer ate bark throughout the year. Even though the amount of the bark eaten per capita was small, the high density of deer may possibly cause serious bark damage. Barking can be one of the main causes of the mortality ofPicea jezoensis Sieb. et Zucc. var.hondoensis (Mayr) Rehder andAbies homolepis Sieb. et Zucc. in Mt. Ohdaigahara. A part of this study was presented at the 105th Annual Meeting of the Japanese Forestry Society (1994).     

Cytokeratin‐positive folliculo‐stellate cells in chicken adenohypophysis

Folliculo‐stellate (FS) cells are non‐endocrine cells found in the adenohypophysis and are identified in many animals by the S100 protein marker. Although keratin is another FS marker in several animals, there is no information on localization of keratin in the avian adenohypophysis. In this study, localization of cytokeratin in chicken adenohypophyseal cells was investigated immunohistochemically. Basic cytokeratin (bCK)‐positive cells were arranged radially in the cell cords with their cytoplasmic processes reaching the basal lamina. The cell bodies encircled a follicle in the center of the cell cord. Furthermore, the bCK‐positive cells were also S100B‐positive. Growth hormone, prolactin, adrenocorticotrophic hormone, and luteinizing hormone β‐subunit did not co‐localize with the bCK‐positive cells. In addition, the bCK‐positive cells had a laminin‐positive area in their cytoplasm. Transmission electron microscopy observed agranular cells equipped with several microvilli that encircled a follicle. These results indicate that bCK‐positive cells in the chicken adenohypophysis may be a predominant FS cell population and produce laminin. It is suggested that they function as sustentacular cells to sustain the adjacent endocrine cells and the structure of the cell cords in the chicken adenohypophysis.     

Estradiol inhibits hepatic stellate cell area and collagen synthesis in the chicken liver

Hepatic stellate cells (HSCs) are the main collagen‐producing cells in the liver. The HSC area and amount of collagen fibers are different between male and female chickens. This study was performed to confirm the effect of estradiol on collagen synthesis in the growing chicken liver. Blood estradiol levels in chicks were compared at 4 and 8 weeks of age, and the collagen fibril network in liver tissue was observed at 8 weeks by scanning electron microscopy. Intraperitoneal administrations of estradiol and tamoxifen to male and female chicks, respectively, were performed daily from 5 to 8 weeks of age. The areas of HSCs and collagen contents were measured in the liver tissue. The blood estradiol level was higher in females than in males, and the collagen fibril network was denser in males than in females at 8 weeks of age. Estradiol administration in males induced decreases in the HSC area and collagen content of the liver. Conversely, tamoxifen administration in females induced an increase in the HSC area but did not facilitate collagen synthesis. Based on these results, estradiol inhibits the area and collagen synthesis of HSCs in the growing chicken liver under normal physiological conditions.