IN earlier work on the mammotrophic activity present in women's urine, intact weanling male mice of a Swiss Schneider strain (SAS3) were used as test objects (Hadfield and Young, 1956a, b). The injection of urine or urine extracts into such mice stimulated development of the mammary glands as shown b.N, extension of the duct system and an increase in the number of end buds or clubs.Similar activity was possessed by oestrone and by prolactin.Gradually the commercially-bred SAS3 mice lost sensitivity under test and they were replaced by mice of the inbred A2G strain. This was the only other strain out of 37 examined in which the mammary glands of weanling males were responsive to both prolactin and urine (Young, 1957).The mammotrophic activity in urine was believed to be due to protein hormone(s) from the hypophysis and it was therefore considered advisable to continue the work using hypophysectomized mice given partial steroid replacement JA7,ith oestrone and progesterone (Hadfield, 1957).The mammary glands of the hypophysectomized immature males of the A2('Astrain failed to respond to oestrone plus progesterone or to prolactin or urine alone but responded to combined treatment with all three hormones or to urine given with oestrone and progesterone. Serum from normal women was also effective in combination with the two steroid hormones (Hadfield and Young, 1958).The present paper reports a somewhat unsuccessful attempt to extend this work.We had hoped to be able to measure the mammotrophic activity in human urine and blood, in order to establish normal values and study variations. This has, however, proved virtually impossible with the test system used, because the responsiveness of the mice to prolactin has diminished. Neat urine is now seldom effective in the test and though mammotrophic activity has again been demonstrated unequivocallv in women's serum, the amount can be estimated onl very roughly. MATERIALS AND METHODS AnimalsMice of the A2G strain were bred in our laboratories from random matings of trios replenished every 3-4 generations with fresh stock from the Laboratorv Animals Centre.Males were hypophysectomized when 22-26 days old and 10-15 g. in weight. These were fed on diet 41B supplemented with rolled oats and had access to water throughout. After operation they were kept at a temperature of 78' C. The mice
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