Volume 50: 275-282, 2001

Detection of Estrogenicity by Bioassay on the Mouse Mammary Gland in Vivo

J. ŠKARDA

Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic

Received April 27, 2000
Accepted October 13, 2000


Summary
The wide chemical diversity of estrogenic compounds precludes an accurate prediction of estrogenic activity on the basis of chemical structure or radioimmunological assay and thus requires that the potency of these compounds is defined by bioassay. The mammary duct growth response in intact prepubertal and adult gonadectomized female and male mice of the C3H/Di strain was used to assess the estrogenicity of synthetic compounds or their derivatives. The vehicle for tested compounds should be free of estrogenic and other hormonal effects. Olive oil or sunflower oil exerted estrogenic activities and were thus unsuitable as vehicles for the tested compounds. The absence of estrogenic activity, high solubility of different steroid hormones, and the low incidence of the inflammatory reactions at the injection site were achieved by using a vehicle containing benzyl alcohol, benzyl benzoate, butylhydroxyanisole, butylhydroxytoluene, ethyl oleate and ethanol. The bioassay was primarily designed to examine the effect of tested compounds on mammogenesis. The duration of hormone treatment was chosen to be long enough for induction of duct growth but too short to induce lobuloalveolar differentiation. Females were treated for 10 days, males for 15 days. The proportional volume occupied by mammary epithelial structures was estimated by the modified Chalkley's technique. The mean coefficient of variation of quantitative evaluation of 10 different mammary glands obtained by two operators varied between 3.2 and 17.4 %. The mean coefficient of variation of quintuplicate determinations of each mammary gland by one operator was 10.1 %, and 11.1 % by the other. The correlation coefficient between results of two operators was 0.994. Estrogens are primarily defined by their ability to increase the mitotic activity of female secondary sex organs. However, our results have shown that progesterone alone, if administered in a high dose, stimulates mammary growth in both intact prepubertal and OV-X female mice similarly as the synthetic progestatial steroid norethindrone with inherent estrogenic properties. In contrast, progesterone alone had no effect, in young intact or adult castrated males, but norethindrone did stimulate mammary growth. These results demonstrated that the mammary gland of males is a suitable model for estrogen screening.


Key words
Mammary gland · Mouse · Estrogenocity · Bioassay · Progesterone

Reprint requests
Josef Škarda, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Přátelství 560, 104 00 Prague 10-Uhříněves, Czech Republic, e-mail: skarda@iapg.cas.cz


PHYSIOLOGICAL RESEARCH
© 2001 by the Institute of Physiology, Czech Academy of Sciences

ISSN 0862 - 8408

Issue 3