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Adrenal Tumors and Other Pathological Changes in Reciprocal Crosses in Mice II

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Adrenal Tumors and Other Pathological Changes in Reciprocal Crosses in Mice II Adrenal Tumors and Other Pathological Changes in Reciprocal Crosses in Mice II. An Introduction to Results of Four @eciprocai• 11' @rosses* G. W. WOOLLEY,tM.M. DICKIE,ANDC. C. LITTLE (Roscoe B. Jackson Memorial Laboratory, Bar Harbor, Maine) This report is to serve as an introduction to hy brief review of a few of the characteristics of the bridization studies which have been undertaken inbred strains used as parental strains in these re concerning adrenal cortical tumors and related ciprocal crosses may help to evaluate the results pathological conditions. It is planned that reports obtained in this now extensive pilot experiment. similar to that for the reciprocal DBA and CE hy It has been customary to characterize inbred brids (27) will also be made. strains according to certain factors, e.g. , mammary Hybridization studies have been used in many tumor incidence, susceptibility or resistance to the types of experiments with animals and with plants. mammary tumor inciter (MTI), response to Such experiments were of the type Mendel used gonadectomy, and incidence of various other types and on which he based his principles of unit inher of tumors. Some of these characteristics will be itance. Hybridization experiments with animals briefly considered, where possible, for the inbred can be used not only to determine chromosomal in parental strains used. (a) Strain DBA has a mod heritance, but also for purposes of evaluating other erate mammary tumor incidence ; following gon types of influences on the offspring, e.g., maternal adectomy, nodular hyperplasia of the adrenal cor influence influential in breast tumor occurrence in tex develops, and the accessory reproductive or mice. gans become feminized (10, 28, 29, 30). (b) Strain The effects of gonadectomy on certain inbred CE has no mammary tumors but is thought to be strains of mice have already been reported in de susceptible to the MTI. Old females in this strain tail (9, 10, 19, 23, 28—36),as well as some intensive sometimes develop granulosa-cell tumors of the studies on the pituitary (3, 5, 26) and the uterus ovary. Gonadectomy causes adrenal cortical car (2) of some F1 reciprocal crosses. Recently, the cinomas to develop, and concomitant with the tu findings on the reciprocal cross of strains DBA and mor is feminization and/or mascutinization of the CE have been reported (27). With this as a pat accessory reproductive organs (81—35).(c) Strain tern, it is possible to indicate how the other recip CS7BL/10 does not have mammary tumors and is rocal crosses thus far investigated compare with relatively resistant to the MTI. Gonadectomy this series. causes the “classical―castrate picture, no major Primarily, the entire series was undertaken to changes in the adrenals, and the accessory repro determine the effect of inheritance on the response ductive organs typically remain small and imma of the adrenals of the F1 animals to gonadectomy. ture. (d) Strain A/Wy has a low mammary tumor However, certain other factors could not be neg incidence in virgin females but a high (85—90per lected and have been included in the results. A cent) incidence in breeding females (17). The re sponse to gonadectomy in this strain is very simi * This work has been aided by grants to the Jackson Me morial Laboratory from the Commonwealth Fund, the Anna lar to the response of strain CS7BL/l0.' (e) Fuller Fund, the Jane Coffin Childs Memorial Fund for Medi In strain CSH, the mammary tumor incidence cal Research,the National CancerInstitute ofthe National In is high (93—96per cent) in both virgin and breeding stitutes of Health, Public Health Service, and the American females.' The adrenal response to gonadectomy Cancer Society. is similar to that of strain DBA.' (f) The yellow t Head, Division of Steroid Biology,Sloan-KetteringInsti tute, New York, and Visiting Research Associate, Jackson strain was introduced into the experiment because Memorial Laboratory. of its interesting genetic constitution and its dem Received for publication October 17, 1952. 1 Unpublished data. 231 Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 1953 American Association for Cancer Research. 232 Cancer Research onstrated ability to modify the incidence of mam RESULTS mary and lung tumors. Homozygous yellow ani GENERAL COMMENTS ON THE ANIMALS OF mats die early in embryonic life; therefore, all yet THE VARIOUS CROSSES low animals are heterozygotes, and matings to Both body size and activity separated the F, ani these yellows produce two classes of offspring that, mats into two main groups : those in which CE is phenotypicatly at least, differ from each other by one of the parental strains and those without any only one gene. It is known that yellow outcross CE inheritance. animals often become relatively obese compared DBA x CS7BL/1O and the reciprocal F, ani to other animals. The response to gonadectomy is mats did not seem much larger in body size than not known. strain CS7BL/1O animals but were larger than strain DBA/2 mice. As a whole, CS7BL X DBA MATERIALS AND METHODS and the reciprocal animals were only moderately The experimental methods may be divided into two sec active and not skittish or nervous. tions : (a) the manner in which the crosses were made and (b) A X C3H and the reciprocals were fairly short the actual experimental procedure used for collecting data on the animals raised. of body, i.e., they did not appear to have so great a) Five series of crosses were made as follows: a body length as any other F1 animals. In activity 1. DBA/2Wy 9 X C57BL/lOd' and the reciprocal they were the most calm and quiescent of all the C57BL/10 9 X DBA/2ci'. F, animals raised. 2. DBA/2 9 x CE/Wyd' and the reciprocal CE 9 X On the other hand, DBA X CE and the recip DBA/2O'. rocal were long and rangy and the most active of 3. CE 9 X C57BL/lOd' and the reciprocal C57BL/ 109 XCEO―. all the F, animals. CE X CS7BL and the recipro 4. A/Wy 9 X C3H/Jaxc? and the reciprocal C8H 9 X cats, although somewhat calmer, were large and Ad'. rangy like the other CE hybrids. @ 5. YEL x CEd' which produced two types of F1 off The yellow animals became moderately obese spring, yellow and agouti. No reciprocal cross was made. and were very docile, while their nonyellow sibs were very similar to the other CE hybrids in size In designating crosses hereafter in the text, the female par and activity. ent is always listed first. b) The experimental procedure differs very little from that Some indication of these size differences may be used in previous studies on the response of various types of ascertained by comparing body weights of the ani mice to gonadectomy. Animals were gonadectomized at birth mats of the various F, groups (Tables 1 and 2). to 8 days of age and then allowedto mature and age without further treatment. Intact virgin females and unmated males ADRENALS were raised as controls and aged with no treatment. All ani mals, experimentand control, weremaintained under uniform INTACT A@mw.@s environmental conditions and fed Purina Fox Chow and water Size.—The adrenals of alt intact animals were ad libitum. measured, and total adrenal, cortical, and medul One of each of the intact females, males, gonadectomized lary size were computed. These measurements of females, and males was autopsied at 15 days, 1 month, and at monthly intervals up to 24 months of age and beyond to obtain the adrenats of intact females and intact males of a developmental series. An exception to this was the A X CSH each reciprocal cross were compared and analyzed and C3H X A set of crosses which were autopsied at bimonthly statistically (Tables 3 and 4). Average measure intervals beginning at 2 months of age. There were several gaps ments of each pair of crosses were determined, and in the YEL x CE series that were not filled, because the ani these were compared with all other pairs of crosses. mals were killed in the fire of 1947 and the cross was not started again. The results of these computations are given in Tissues regularly observed and saved for histological study Tables 5—10. were the submaxillary gland, both adrenals (which were serial From this statistical analysis it can be observed sectioned), the ovaries (the left was serial sectioned) or testis that the differences between pairs of crosses are (when present), uterus, vagina, the accessory sex organs (in eluding seminal vesicles and prostates) of the males, the thy greater and more significant than between recipro roid, and the pituitary. Other organs with any observed gross cats. The animals with the largest adrenals were abnormalities were also saved. All tissues were ordinarily fixed YEL x CE, CE x CS7BL and DBA X CE and in Tellyesnicky's fluid, cut at 8 gsand stained with hematoxylin their reciprocals, i.e., those with a CE parent. and aqueous eosin. The pituitary glands, when abnormal, were fixed in modified Zenker's solution, serially sectioned at 4 p, Structure.—Changes observed in the adrenals of and differentially stained. The skins of all animals were fixed the intact animals can be divided into (a) those in and saved so that the mammary glands could be studied. the cortical elements and (b) those in the medul Measurements on all tissues, where indicated, were made lary area. with an ocular micrometer as described previously (27). Statis Subcapsular “A―cellsnoted in DBA X CE and tical analyses, the significance of the mean difference of these various measurements, were determined by Fisher's method CE x DBA F, females by 3 months (27) were (11) for small samples using available paired observations.
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