Kazuko Aoki

Life-span Studies on Spontaneous Tumor Development in the Medaka (Oryzias latipes)

A total of 961 medaka, separated chronologically from the first to the fifth year of life, were examined for spontaneous tumor development. While no liver tumors were found in either male or female medaka under the age of 1 year and the incidence in 2‐year‐old fish was relatively low (males 1.9% and females 1.7%), they became more common with advancing age. The incidence was higher in females than in males from 3 to 5 years of age, reaching 7.1% in 5‐year‐old female stock. These liver tumors included a total of 12 adenomas and 9 hepatocellular carcinomas. The hepatocellular carcinomas were histologically well differentiated and were all observed in female medaka. Spontaneous tumors occurring in organs other than the liver were rare and sporadic. Four squamous cell carcinomas, 5 melanomas and 4 lymphosarcomas were observed with no sexual or pronounced age bias being evident. The squamous cell carcinomas developed in the surface epithelium with local invasion into the dermis. Melanomas occurred in the abdominal cavity and demonstrated systemic invasion into various parts of the body. Three out of the 4 lymphosarcomas arose from the inner part of the operculum suggesting that these tumors were of thymic origin. They also showed extensive invasion. The data indicate a particular susceptibility of older female medaka to liver but not other tumor development.

Age and strain dependence of O6-methylguanine DNA methyltransferase activity in mice

The activity of the DNA repair protein O6-methylguanine DNA methyltransferase (MT) was compared in liver extracts from female ICR and male C57BL/6 mice at various ages (3-130 weeks old). Similar patterns of overall enzyme activity were observed in both strains with O6-MT activity being relatively low in young mice (3 or 8 weeks old). However, the activity significantly increased after adolescence (middle age), thereafter decreasing with old age (over 100 weeks old) to a level equivalent to that found in young mice. In an additional strain difference study, O6-MT activities in liver extracts from 4 strains of mice were compared at 5 and 30 weeks of age. Although a similar age-associated increase of enzyme activity in adolescence was confirmed in all 4 strains investigated, the closed-colony ICR mice differed from the inbred strains in demonstrating significantly higher levels of O6-MT activity in females than in males. However, the same tendency was also observed in a comparison of the sexes in 30-week-old C3H/HeN, C57BL/6 and BALB/c mice.

Mutation induction by different dose rates of gamma rays in near-diploid mouse cells in plateau- and log-phase culture

Induction of mutation to 6-thioguanine resistance was studied in cultured near-diploid mouse cells (m5S) in plateau and log phase after exposure to gamma rays at dose rates of 30 Gy/h, 180 mGy/h, or 13 mGy/h. In plateau-phase culture, lowering the dose rate from 30 Gy/h to 13 mGy/h resulted in an increase in cell survival and a marked decrease in induced mutation frequency. On the other hand, in the log-phase culture, the magnitude of the dose-rate effects was not as marked as in the plateau-phase culture, particularly within a dose range below 5 Gy. These results, together with those indicating the inverse dose-rate effects in growing mouse leukemia cells (Radiat Res. 115, 273-280, 1988), demonstrate the significant influence of cell growth that takes place during protracted irradiation, particularly for the induction of mutation.

Age dependence of O6-methylguanine-DNA methyltransferase activity and its depletion after carcinogen treatment in the teleost medaka (Oryzias latipes)

O6-Methylguanine-DNA methyltransferase (O6-MT) is considered to play an important role in the repair of DNA lesions induced by alkylating carcinogens in a wide range of animals. The activity of O6-MT was compared in liver extracts from the teleost medaka (Oryzias latipes) at various ages (3-5 years old) reared under natural conditions. O6-MT activity decreased significantly with advancing age. When medaka were exposed continuously to the alkylating agent methylazoxymethanol (MAM) acetate at levels of 0.1, 0.15 and 0.3 ppm in water, O6-MT activity was markedly reduced from days 1 to 7, with a slight increase thereafter. Furthermore, when fish were exposed to MAM acetate at levels of 1-2 ppm for 1 h and then maintained in normal tap water, O6-MT activity remained suppressed for 2 weeks, followed by a partial recovery.

METAL BALANCE SHIFT INDUCED IN SMALL FRESH WATER FISH BY SEVERAL ENVIRONMENTAL STRESSES

Balance of essential elements in organisms might be changed by environmental stresses. Small fresh water fish, Medaka, was burdened with X-ray irradiation (total dose: 17Gy), keeping in salty water (70% NaCl of sea water) and keeping in metal containing water (10ppm of Cr and Co). These stresses are not lethal doses. Essential elements in liver, gall bladder, kidney, spleen, heart and brain in the stress-loaded fish were measured by PIXE method and compared with a control fish to determine the effect of the stresses. Various changes of the elemental contents were observed. Effect of X-ray irradiation was the smallest among the stresses. Relatively high content elements such as P, S, Cl and K were hardly affected with the stresses examined in this work. The effect of Cr on the metal balance seems to be larger than the other stresses. As PIXE method can analyze many elements in a small sample simultaneously, change of elemental distribution in small organisms induced by environmental stresses can be determined readily.

MEDAKA, A USEFUL EXPERIMENTAL SYSTEM FOR CHEMICAL AND ENVIRONMENTAL CARCINOGENESIS

Publisher Summary This chapter describes the application of the medaka in chemical and radiation carcinogenesis. The medaka, Oryzias latipes, is a convenient system to study chemical and environmental carcinogenesis at molecular, cellular, and tissue levels. It is economical and easy to breed. Chemicals can be administered as solution added to the water in aquaria or be injected into the fish body. The fish is sensitive to water-soluble carcinogens including diethylnitrosamine, methylazoxymethanol (MAM) acetate, and N-Methyl-N´-nitro-N-nitrosoguanidine. The presence of cytochrome P-450 system indicates a possibility to use this fish as a test system for other carcinogens that need metabolic activation. Tumor formation can frequently be detected from the body surface as experienced in melanomas and hepatomas. The small size of the fish permits routine serial sectioning for histological examination.