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The Female Mammary Gland in Two Strains of Albino Mice * Lydia M

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The Female Mammary Gland in Two Strains of Albino Mice * Lydia M A COMPARATIVE STUDY OF THE LIFE HISTORY OF THE FEMALE MAMMARY GLAND IN TWO STRAINS OF ALBINO MICE * LYDIA M. GIBSON (FTo~the Department of Pathology, Yale University School of Medicine) In searching the literature nothing could be found concerning the life history of the mammary gland in mice, although much work has been done on other rodents. Since the mouse is used so extensively in cancer research as an experimental animal, and since cancer of the mammary gland is a well recognized entity in certain strains of mice, it was thought that it would be interesting to make a comparative study. To do this, two strains of mice were necessary, one showing a low incidence of mammary cancer in the females, and the other showing a high incidence. The material presented here represents an attempt to work out the development and distribution of mammary tissue in two such strains of mice. The study is a preliminary one, and the material is limited, but it was decided that should the two strains exhibit any remarkable differences, a more extensive survey might prove both interesting and profitable. The high tumor incident strain has been designated through- out as SPTM, and the low tumor incident strain as Loomis 1. Bagg strain was abandoned because of lack of material. Data concerning the animals used in this study are found pre- ceding the plan of work. PURPOSE. By gross and microscopic study (1) to trace the development and distribution of mammary tissue in female mice; (2) to compare two strains: (a) one having a low mammary car- cinoma incidence (no strain that has been studied carefully has been known to be tumor free) and (b) a high spontaneous mam- * A thesis submitted to the Faculty of the Yale University School of Medicine in candidacy for the degree of Doctor of Medicine. 570 mary tumor strain; (3) to find, if possible, when deviations from the normal begin to occur, and to trace these deviations to the developed neoplasm. MATERIAL:The material for this work is drawn from the fol- lowing sources : I. Control Strains: (1) Bagg (low mammary tumor). Re- ceived one pair of breeders March 20, 1928. Albinos from stock of albino strain established 1913. Mice received from dealer in Ohio, inbred by brother and sister mating, no new blood added; breeders selected for vigor and breeding qualities; no other infor- mation available except that these mice have shown a low in- cidence of mammary carcinoma. (This refers to female mice). (2) Loomis 1. Four males and ten females received April 12, 1928; ten females and two males received May 25, 1928; three females and one male received April 12, 1928; donated by Dr. Lewis, Phipps Institut,e, Princeton. Original stock from Cam- den, N. J. Sixteenth generation of brother and sister matings (G. stock). No mammary carcinoma ever observed in this strain. II. Spontaneous Mammary Carcinoma Strain: Twenty-one females and two males received March 22, 1928, from the State Institute, Springville, N. Y. Thirteen females and eight males were received on July 23, 1928. Original stock obtained from Abbie Lathrop, Cranby, Mass. No information was secured except that the mice were from high tumor stock. None were identified as high or low representatives or in any other way. They were bred at random by M. C. Marsh at Springville, N. Y. At least nine inbred strains were carried out, before finding the one that is the present high tumor strain (known as strain 3). The mice used in this experiment were offspring of this high tumor strain. Tumors observed in these mice were predomi- nantly mammary carcinoma, and these neoplasms have been observed only in females. This is the twenty-fifth generation; the last few generations have yielded 80 per cent tumors. Sarcomas are rarely observed in these mice. Tumors develop in the fifth to*the nineteenth month. The highest development is reached in a year and a half to two years. No mammary car- cinoma has ever been observed in male mice of this strain. 572 LYDIA M. GIBSON Diet: All mice were fed on an adequate mixed diet, and a sup- plementary feeding of milk powder was given to the nursing mothers. Breeding: Strains have been kept separate and inbreeding carried on. Details of exact data concerning mating, gestation, weaning, and age will be found in the individual records. Plan of Work: Using histological preparations for study: Control Spontaneous Tumor Strain 1. Trace the development of 1. Procedure same as control. the mammary glands from the first indications of the milk line in the embryo to the development of the nipples with the underlying immature gland tissue. This requires sectioning embryos of different stages in their development. 2. Series of sections showing 2. Procedure same as control. the state of glands at birth. 3. State of glands at varying 3. Procedure same as control. age periods (one month, two months-puberty, three months-mature) . 4. Lactating breast. 4. Procedure same as control. 5. Adult resting breast. 5. Procedure same as control. 6. Series of sections of breast 6. Series of sections showing, tissues from adult females, different types of mastitis breeders and non-breeders, and a series similar to the from the active reproduc- control, concerning atrophic tive period to the atrophy of changes. In addition, a ser- old age. ies showing the occurrence and distribution of neo- plastic growths at different age intervals. FEMALE MAMMARY GLAND IN TWO ALBINO MICE 573 PROCEDURE.1. For the embryos, the animals were mated. The copulatory plug read and the date recorded, the animals were killed on the tenth, fourteenth, nineteenth days from the dis- covery of the plug. Embryos were removed from the uterus and fixed in 10 per cent formalin. After imbedding in paraffin, serial sections, 10 micra thick, were made. Several attempts to secure a nineteen-day embryo in the control strain were made, but in one case the uterus was opened and the embryos were partially absorbed, and in three other cases the animals refused to mate; in the fourth case the plug was read, but due to unfore- seen circumstances the animal came to term and delivered. Time shortage prevented further mating, and this embryo had to be omitted. 2. Skip serial sections, 10 micra thick, every tenth section used on all animals sectioned from birth through three-month devel- opment. Animals at birth were fixed in 10 per cent formalin, decapitated to cut down extra expense of sectioning, and cross sections through the whole body were made. From the tenth day to the third month, the following procedure was carried out: For the tenth day post-natal animals, the skin was removed by slitting it up the center of the back, cutting it at the root of the neck and at the base of the tail, and peeling it off glove-fash- ion from the body. The deposits of fat in the inguinal and tho- racic regions came with it. This skin was then spread out on a blotting paper and pinned in position, labelled, and fixed in 10 per cent formalin. Drawings were made of the location of the nipples and the fatty deposits. When the skin was fixed, it was divided by cutting down the mid-line of the ventral surface. One-half was used for sectioning and the other half was saved. The band of tissue between the thorax and the abdomen, which is free from nipples and fat deposit, was discarded. The tenth day post- natal animals were so small that the whole portion could be sec- tioned. But the areas so obtained in the first, second and third month development were too large to section in one piece, so the thoracic and inguinal deposits were divided into three portions and sectioned. Rough drawings were made for identification, e.g. thoracic deposit labelled th. A, B, C and ing. A, B, C. By 37 574 LYDIA M. GIBSON this scheme, which worked out fairly well, the whole deposit, in- cluding the nipples, was sectioned. For the older mice (six months onward), single sections were made from tissues taken either from the thoracic or inguinal deposits, as indicated in the notes.' It was not thought neces- sary to section the older mice so extensively, since this procedure was carried out thoroughly up to maturity, and a fair conception of the distribution was gleaned from the study of these slides. Note: Measurements were made on the embryos from the head end to the tail end. The embryo was laid on a centimeter ruler and its length was ascertained. Measurements Control Spontaneous Tumor Mice 10 day embryo L.I.G. 3 mm. 10 day embryo SPTM 3 mm. 14 day embryo L.I.G. 10.5 mm. 14 day embryo SPTM 11 mm. 19 day embryo L.I.G. 19 day embryo SPTM 15 mm. At birth L.1.C:. 15 mm. At birth Animals were not weighed or measured after this period. Slides were studied carefully and written up. Photographs were made of interesting areas and are included with the data. EMBRYONICDEYELOPMENT: The mouse follows the general plan of mammalian development. The mammary streak and the mammary line doubtless appear shortly after the tenth day of embryonic life. I was unable to identify the line in either of the embryos of this age, but remnants of the line are still recognizable in embryos of fourteen days. This line of heaped up epithelial cells extends from the root of the neck to the groin on each side of the body, lateral to the midline. Along this line the future nipples develop.
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