Enzyme Immunoassay of 17β-Estradiol, Estrone Conjugates, and Testosterone in Urinary and Fecal Samples from Male and Female Mice

ELISA measures of 17β-estradiol, estrone conjugates, and testosterone were adapted for fecal and urinary samples from laboratory mice. We will report on validations of these assays and data from interacting males and females. Unconjugated gonadal steroids were consistently measurable in urine and feces of both males and females. Females that were parturient following insemination excreted relatively low levels of urinary testosterone compared to non-parturient females. The results are consistent with evidence that elevated androgens and estrogens are incompatible with intrauterine implantation of fertilized ova, and suggest that steroids in male urine could contribute to pheromonal action. These methods permit repeated noninvasive measurement of steroid activity in this species.

Key words:

Mice - Testosterone - 17β-Estradiol - ELISA - Pregnancy - Excretions - Bruce Effect

References

• 1 Carroll J B, Abbott D H, George L M, Hindle J E, Martin R D. Urinary endocrine monitoring of the ovarian cycle and pregnancy in Goeldi’s monkey (Callimico goeldii). .  J Reprod Fertil. 1990;  89 149-161
• 2 Graham L H, Goodrowe K L, Raeside K I, Liptrap R M. Non-invasive monitoring of ovarian function in several felid species by measurement of fecal estradiol-17β and progestins.  Zoo Biology. 1995;  14 223-237
• 3 Lee J V, Whaling C S, Lasley B L, Marler P. Validation of an enzyme immunoassay for measurement of excreted estrogen and testosterone metabolites in the White-Crowned sparrow (Zonotrichia leucophrys oriantha).  Zoo Biology. 1995;  14 97-106
• 4 Munro C J, Stabenfeldt G H, Cragun J R, Addiego L A, Overstreet J W, Lasley B L. Relationship of serum estradiol and progesterone concentrations to the excretion profiles of their major urinary metabolites as measured by enzyme immunoassay and radioimmunoassay.  Clin Chem. 1991;  37 838-844
• 5 deCatanzaro D, MacNiven E. Psychogenic pregnancy disruptions in mammals.  Neurosci Biobehav Rev. 1992;  16 43-53
• 6 Bruce H M. A block to pregnancy in the mouse caused by the proximity of strange males.  J Reprod Fertil. 1960;  1 96-103
• 7 deCatanzaro D, MacNiven E, Ricciuti F. Comparison of the adverse effects of adrenal and ovarian steroids on early pregnancy in mice.  Psychoneuroendocrinology. 1991;  16 525-536
• 8 Harper M JK. Estrogenic effects of dehydroepiandrosterone and its sulfate in rats.  Endocrinology. 1969;  84 229-235
• 9 deCatanzaro D, MacNiven E, Goodison T, Richardson D. Estrogen antibodies reduce vulnerability to stress induced failure of intrauterine implantation in inseminated mice.  Physiol Behav. 1994;  55 35-38
• 10 deCatanzaro D, Muir C, O’Brien J, Williams S. Strange-male-induced pregnancy disruption in mice: Reduction of vulnerability by 17β-estradiol antibodies.  Physiol Behav. 1995;  58 401-404
• 11 Greenwald G S. Luteolytic effect of estrogen on the corpora lutea of pregnancy of the hamster.  Endocrinology. 1964;  76 1213-1219
• 12 Burdick H O, Whitney R. Acceleration of the rate of passage of fertilized ova through the fallopian tubes of mice by massive injections of an estrogenic substance.  Endocrinology. 1937;  21 637-643
• 13 Suginami H. Endocrine regulation of early embryonic development and implantation.  Horm Res. 1995;  (Suppl) 44 1-3
• 14 Pfaff D W. Estrogens and brain function.  New York:; Springer-Verlag, 1980
• 15 Bruce H M. Effect of castration on the reproductive pheromones of male mice.  J Reprod Fertil. 1965;  10 141-143
• 16 deCatanzaro D, Smith M, Muir C. Strange-male-induced pregnancy disruption in mice: Potentiation by administration of 17β-estradiol to castrated males.  Physiol Behav. 1995;  58 405-407
• 17 Zacharias R, deCatanzaro D, Muir C. Novel male mice disrupt pregnancy despite removal of the vesicular-coagulating and preputial glands.  Physiol Behav. 2000;  68 285-290
• 18 Reynolds E. Urination as a social response in mice.  Nature. 1971;  234 481-483
• 19 Drickamer L C. Rates of urine excretion by house mouse (Mus domesticus): Differences by age, sex, social status, and reproductive condition.  J Chem Ecol. 1995;  21 1481-1493
• 20 deCatanzaro D, Zacharias R, Muir C. Disruption of early pregnancy by direct and indirect exposure to novel male mice: Comparison of influences of preputialectomized and intact males.  J Reprod Fertil. 1996;  106 269-274
• 21 Rajendren G, Dominic C J. Effect of cyproterone acetate on the pregnancy-blocking ability of male mice and the possible chemical nature of the pheromone.  J Reprod Fertil. 1988;  84 387-392
• 22 deCatanzaro D, Baptista M AS, Spironello-Vella E. Administration of minute quantities of 17β-estradiol on the nasal area terminates early pregnancy in inseminated female mice.  Pharmacol Biochem Behav. 2001;  69 503-509
• 23 Kemeny D M. A practical guide to ELISA.  Oxford, England:; Pergamon Press, 1991
• 24 deCatanzaro D, Muir C, Sullivan C, Boissy A. Pheromones and novel-male-induced pregnancy disruptions in mice: Exposure to females is necessary for urine alone to induce an effect.  Physiol Behav. 1999;  66 153-157