Spontaneous Basophilic Tumors of the Pituitary Glands in Gonadectomized Mice*T

Spontaneous Basophilic Tumors of the Pituitary Glands in Gonadectomized Mice*t

MARGARET M. DIcKIE, M.A., AND GEORGE W. WOOLLEY, PH.D.

(Roxcoe B. Jackson Memorid Laboratory, Bar Harbor, Maine)

The presence of spontaneous pituitary tumors following estrogenic treatment (16). Burrows described and lesions following the appearance of adrenal such a tumor in a male mouse following estrogenic treat cortical carcinomas presents a new line of evidence ment (5). Gardner and Strong have shown that the pituitary response to esttogen treatment was strain lim for the close relationship of adrenal cortex and ited and that the strain C57 Black mice were more pituitary disfunctions in experimental animals and susceptible than other strains (16). Pituitary tumors of of these disfunctions to tumor formation and their the above type were observed following implantation of possible inherited tendencies. In man, there are pellets containing an estrogenic substance, diethyl not sufficient data to say conclusively whether the stilbestrol, in strain ce mice in this laboratory (45). adrenal cortex or anterior pituitary abnormalities Castration changes in the pituitaries of rats have are the primary causal agents in the development been described by many workers, Addison (1), Biggart of various syndromes, such as Cushing's syndrome, (2), Guyer (20), and Severinghaus (30). They have (Cushing'sdisease)and adrenal virilism.In fact noted that there is an increase in the number of baso the clinical evidence of these syndromes is so con philes and that these basophiles become vacuolated to fusing that it is difficult to try to ascribe the causal such an extent that they are called â€œsignet ringâ€• cells (30). These changes, however, have not been found to agent to any one endocrine gland and/or its secre any extent in the mouse hypophysis following gonadec tions. Kepler (@) recently has given a critical re tomy (13). view of Cushing's syndrome and its association -Theeffectsofearlygonadectomyinmiceofvarious with Crooke's changes of the basophile cells of the strains have been intensively studied by Woolley, anterior pituitary, i.e. hyalinization of the basophile Fekete, and Little (14, 41, 4@,48). It was observed that cells. He made several postulates about the mecha the strains used reacted differently to gonadectomy nism that might be involved, to be discussed later, Experiments were carried out with several inbred hut did not definitely say that either the anterior strains, namely, JAX, C57 Black, C57 Brown, dba, cc, pituitary or the adrenal cortex was the primary A and CSH.Their resultswillbe summarizedbriefly here. In strains C57 Black, C,57 Brown and A the acces cause. Hyperadrenocorticism, however, usually sory reproductive organs remained small, the submaxil plays an essential part in this syndrome. In the lary gland unstimulated, that is, small and dark; no experimental animals under discussion in the pres obvious changes in the hypophyses or extensive changes ent report it is definite that the adrenal cortical in the a4renals were noted (46). However, the dba and carcinomas precede the gross pituitary abnormali cc mice reacted quite differently. The dba gonadecto ties by several months. The material may be useful mized animals developed nodular hyperplasia of the in helping to elucidate the intimate interplay of adrenal cortex; this modified cortex seemed to be sup pituitary and adrenocortical hormones. plying hormonal stimulation to the organs ordinarily in fluenced by the hormones of the gonads themselves. The A review of the literature indicates that spontaneous accessory reproductive organs in gonadectomized female hypophyseal abnormalities in mice are extremely rare. mice were stimulated and, as histologically determined, Slye found only one abnormality in 11,000 mice cx they were feminized (14). A few mammary tumors also amined (32). Tumors of the mouse hypophysis inter appeared in this experimental group. The pituitaries preted as chromophobe adenomas have been produced were examined and some abnormalities noted but

a This work has been aided by grants to the Roscoe B. changes were not studied in detail at that time. The cc Jackson Memorial Laboratory by the Commonwealth Fund, mice not only developed nodular hyperplasia of the the Anna Fuller Fund, the Jane Coffin Childs Memorial Fund adrenal cortex but, later, adrenal cortical carcinomas, for Medical Research, and the National Advisory Cancer the latter first appearing in the hyperplastic areas. These Council. carcinomas produced both masculinizing and feminizing t Animal tissues were collected previous to the fire of hormones as evidenced by the histological picture of the October 23, 1947 and most of the tissues were saved. Some de submaxillary glands and the accessory reproductive or tailed notes and slides have been missing since that date. gans in both gonadectomized sexes (42, 43, 12). Changes

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Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 1949 American Association for Cancer Research. DIcKIE AND WooLLrnrâ€”Spontaneous Basophilic Tumors 373 in the hypophyses were observed in a few mice but not various animals were removed and sectioned. Most of studied intensively. these tissues were stained with haematoxylin and eosin. After differences in the reactions of the various strains to early gonadectomy became known, it was de GROSS OBSERVATIONS ON THE cided to study reactions after gonadectomy on animals PITUITARIES of the F1 generation of reciprocal crosses of some of these All gross observations on the pituitaries and strains. As the animals were examined it became evident that frequent hypophyseal abnormalities were occurring mammary glands were made with the aid of a dis several months after the appearance of adrenal tumors secting microscope (7 x ocular and 10 X objec in some of the crosses. Concurrent with this, there was tive). very extensive and unusual mammary gland develop The mouse hypophysis lies on the sphenoid bone ment. and is covered by a tough membrane, the intrasel The other data from these crosses will be reported lar dura (Fig. 1). At autopsy the anterior lobe is separately and only the pituitary abnormalities and the usually light pink in color and the intermediate relationships of the mammary glands to these abnor and posterior lobes have an opaque color, the pars malities will be discussed in the present report.

MATERIALS AND METHODS Approximately 800 F1 mice from reciprocal crosses l)etween dba and C57 Black, dba and Ce, ce and C57 Black, and A and C3H strains were gonadectomized from I to 3 days after birth. The mice were then re turned to the nest and allowed to mature and age with out any further treatment. Virgin females and unmated males from these crosses were used for control animals. All animals, experimental and control, were maintained on a diet of Purina fox chow and water ad libitum, and at a temperature of approximately 70Â°F. Animals from both the experimental and control 1. groups were autopsied at monthly intervals from 15 days and 1 month up to 24 months of age and beyond. FIG. 1.â€”Diagrammatic sketches of die mouse pituitary. Gross and histologic observations were made on the A. dorsal view of whole normal gland as seen at autopsy. Pars endocrine glands and accessory reproductive organs for intermedia is lined and pars nervosa is stippled. B. longitudinal comparative studies. The present report is on the macro section of the gland, the residual lumen is between the pars an scopic and histological findings of the pituitary and tenor and the pars intermedia. C. cross section of gland, tv the mammary glands from these animals. Numerous ab third ventricle, d the dura, as the subdural space, am the arach normalities were encountered in the hypophyses of some noid membrane, and b the brain. of the experimental animals but not in the control ani mals. The pituitaries of the control animals and of the nervosa being slightly whiter than the pars inter experimental animals that did not show gross abnormal media. Normal hypophyses in mice measure about ities were fixed in 10 per cent formalin in normal saline, @.16mm.laterally X 1.00 mm. anterioposteriorly. embedded in paraffin, and sectioned at 8 micra. Several In older mice the glands are slightly smaller, longitudinal sections were cut from each pituitary and 1.90 mm. X 0.9 mm. (measurement on micro stained with Mayer's haematoxylin and aqueous eosin. scopic sections). For the study of the abnormal pituitaries a good dif In some of the gonadectomized F1 mice, the ferential staining method was necessary. Trials with a pituitaries were enlarged and nodular. For in number of fixatives in conjunction with various staining stance one abnormal gland measured 3.7 X @.6 methods showed that the mouse pituitary was difficult to stain differentially, as compared to the rat or the mm. Two kinds of nodular areas were noted in hamster. A method was finally developed using either a these glands. The first type was a well defined modified Zenker-formol solution or a modified Bouin so opaque nodule or lesion at the periphery of the lution for fixation. The sections were stained with a corn anterior lobe that did not alter the normal contour bination of Mallory's and McFarlane's triple staining of the gland (Fig. @).The second type was a pro methods (1 1). Abnormal glands were cut in cross section truding hemorrhagic nodule (Fig. 3). These nod and serially sectioned at 4 micra. ules could be found at either or both tips of the A few normal pituitaries beyond 12 months of age anterior lobe and sometimes were so extensive that were fixed and stained in the same way as the abnormal they pushed the intermediate and posterior lobes pituitaries for purposes of comparison. The mammary glands were left on the skins of the aside but did not seem to invade them. Char animals and were fixed in modified Tellyesniczky's fluid. acteristically they gave the impression of penetrat These glands were then studied with the aid of the dis ing deeply into the anterior lobe. secting microscope and some representative glands from These nodular areas occurred after 14 months of

age, sometimes both kinds in one gland, sometimes The alveolar development that was found along only one. Table 1 shows the data on the eight F1 with the hypophyseal changes was very pro crosses that were made. It shows that there are nounced; the alveoli were large and sometimes the strain variations and emphasizes the fact that the ducts were concealed by the alveolar growth. This pituitary tumors occur at 14 months of age or growth was similar in gonadectomized animals, later. In all cases the individuals that showed both females and males. The only difference was hypophyseal tumors had well developed adrenal that all five pairs of mammae were present in the tumors. The table does not show whether each gonadectomized females, while in the castrated a(lrenal possessed a tumor, but includes all the males some of the glands were absent, probably animals that had one or more adrenal tumors. because the rudiments for some of the glands were never formed.

MICROSCOPIC OBSERVATIONS ON THE PITUITARIES The Opaque Nodules.â€”The opaque nodules that occurred in the hypophyses of the experimental animals were not encapsulated. They were corn posed of densely arranged cells. The cells were mainly of two types : (a) small cells, closely packed in disorderly arrangement @ L81 3. and, (b) very large cells arranged in small acinar

FIG. 2.â€”Diagram of the mouse pituitary showing possible clusters separated from one another by delicate sites of the opaque nodules. reticular fibers. Both types of cells could be found in one opaque nodule, or there might be only one FIG. 3.â€”Diagram of the mouse pituitary showing a large hemorrhagic nodule that has pushed the pars intermedia aside type in an opaque nodule. slightly. The small cells had light staining cytoplasmic basophilic granules and remnants of a Golgi ap GROSS @)BSERVATIONS ON THE paratus comparable in form to the negative image MAMMARY GLANDS of the Golgi apparatus in the normal mature Concurrent with the gross hypophysea.l changes, basophile (i.e., a doughnut shaped negative image there was very extensive alveolar development of associated with basophiles). The nuclei of these the mammary glands. It has been reported pre cells were slightly different from those found in viously that following the appearance of adrenal normal anterior lobe cells. The chromatin was cortical carcinomas in gonadectomized mice of the found only in a light ring at the periphery and cc strain there was some mammary development, there was a large yellow-staining nucleolus as well such as growth and extension of ducts, the appear as several very small fuchsinophile nucleoli. While ance of end buds (4@, 43) and some alveolar devel a few fuchsinophile nucleoli are present in normal opment. Where this latter occurred, it was cor nuclei, no yellow nucleoli were observed in the related with abnormal hypophyseal changes (46). nuclei of the cells of normal glands. Some of these

TABLE 1 FINDINGS FOR GONADECTOMIZED MALES AND FEMALES OF EIGHT F1 CROSSES AT Two AGE GROUPS

AdrenalPituitary.AdrenalPituitarySaxNo. Caoss MICEtumorstumors99OF MICEtumorstumorsNo. or dbaQ XC57d' d'ci@ 8 0 0 16 4 0 C579 Xdbad' 99 8 0 0 16 10 4 d'd' 8 1 0 14 3 0 (lba9 Xced' 99 8 5 0 19 19 9 d'oâ€• 8 1 0 24 24 18 (e9 Xdbad' 99 8 6 0 21 21 12 d'd' 8 1 0 16 16 14 @ ce9 XC57O@ 99 8 3 0 15 14 4 d'd' 8 0 0 15 9 0 c579 Xced' 99 8 4 0 20 20 7 oâ€•Oâ€• 8 2 0 13 11 2 A9 XC3Hd' 99 4 0 0 18 14 4 d'c,@ 4 0 0 13 8 2 C3H9 XAOâ€• 99 4 2 0 10 10 1 d'd'8 40 00 011 119 111 1

The scale in which all cell sketches are reproduced is basophile. There is degranulation of the transitional cells aiid 2 cm. = 0.01 mm. several nuclei are lobular. From 18 months old gonadectomized FIG. 4.â€”Large transitional basophiles and a degranulating male (ce 9 X dbad') F1 hybrid WIK 450. Mag. X439.

câ‚¬lls were partially (legranulated, only a little hypophyses examined, both occurring in the an cytoplasm being found adjacent to the nucleus and tenor lobe. The first and most common type was at the periphery of the cell. lined by thin epithelial cells. The colloid was The cytoplasmic elements of the large cells were usually basophiic, only occasionally being always basophilic and the cytoplasmic elements acidophilic. These cysts were usually very small appeared either finely granular or foamy. The and occurred in clusters or singly. negative image of the Golgi apparatus that was ob The second type of colloid cyst was lined by servable I)y the staining methods used was corn cuboidal ciliated cells (Fig. 9). Some flat non paral)Ie to that found in normal ripe basophiles, ciliated cells were found occasionally. The appear doughnut shaped. Some of these cells had large ance of these non-ciliated cells might be due to a polymorphic and/or lobular nuclei and some cells gradual transformation from non-ciliated to cili were multinucleate. The nucleoli were very large, ated cell type. The colloid in these cysts was al filling almost the entire nucleus. They stained pale ways basophilic. blue and appeared to be vesicular. From the study of the glands it is evident that Occasionally among the above described small cysts are common in the anterior lobe of the pitui and large cells, very small cells were found with a tary. In the present series they were found most clear bluish gray cytoplasm and little or no granu often in control males and gonadectomized females lation. In the nuclei of the cells, the nucleoli were as is shown in Table @.These are by no means ab large and fuchsinophile staining. solute figures since none of the glands of the con There were many variations in the amount of trol animals and only some of the glands of the granulation of the cells that comprised the opaque experimental animals were serially sectioned, nodules. There were no mitotic figures observed in therefore many cysts may have been missed. any of these cells. These cells have been designated Another abnormality was found in several pitui transitional basophiles and the opaque nodules taries of gonadectomized and control animals. shall l)C called focal basophile adenomas. Groups of two to six cells were surrounded by nor tfh acidophiles were absent in the opaque areas mal anterior lobe cells. These encircled cells had while those that were found adjacent to the opaque very large nuclei and the cytoplasm was either nodules were very densely arranged and were very light staining or seemed to be absent. It was somewhat larger than average acidophiles in nor thought that they might be degenerating baso ma! glands. Normal acidophiles ranged from 9.@2 philes, but later they were diagnosed as phago @mcrato 10.41 micra while the acidophiles adja cytes.' cent to the nodules were from 10.17 micra to 13.81 In a castrate male F, (dba 9 X C,57 Blackc,â€•), @1 Inicra in diameter. Other than the size difference months of age six giant cells were found.2 the acidophiles seemed normal. The staining re In WK 706, a @6months old castrate male F1 actions of the a granules seemed normal and no (cc 9 X dbad') the pituitary contained a yellow abnormal nuclei were observed. The negative im nodule. Microscopic analysis showed that this nod age of the Golgi apparatus which forms a cap over ule was composed of cells containing yellow lipo the nucleus in acidophiles also appeared normal. chrome pigment. Close to this area cholesterol The Hemorrhagic Nodutes.â€”The nodules, which clefts separated by flattened epithelial cells were appeared to be hemorrhagic grossly, were also non found. This area of tissue joined both lobes but did capsulated. They frequently contained hernor not invade either lobe to any extent. Such cho rhagic cysts and were surrounded by scattered lesteatomas have been described and do occur oc clusters and strands of abnormal basophilic cells. casionally in the hypophysisâ€• @.In the adjacent In some nodules free blood cells were found amid anterior lobe amyloid degeneration occurred in the strands of abnormal basophiles. In some areas the blood vessel walls. basophiles showed evidence of loss of cohesion. The only abnormality that occurred in the con These basophiles were coarsely granular and the trol series, other than the hyaline cysts, was a negative image of the Golgi apparatus was hyper chromophobe adenoma that was found in a 30 trophied, and filled almost the entire cytoplasmic area. In glands with large hemorrhagic areas and I Diagnoses were made by Miss Elizabeth Fekete of this laboratory. hasophiles, usually no other cell types were pres ent. 2 There seems to be no simple explanation as to why these should have been present in the pituitary. Diagnoses by Miss Other ;lbnormalities.â€”There were several other Elizabeth Fekete of this Laboratory. abnormalities that are worthy of mention and that 3 Cholesteatomas are formed by epithelial cells that were do not fall into the types given above. left behind at the time of the closure of the neural crest and Two types of colloid cysts were noted in the grow slowly throughout life to form these benign tumors (10).

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FIG. 5.â€”Group of acidophiles, chromophobes and a baso tern slide, original negative lost, magnification unknown since phile from a normal section of an abnormal gland. From hy laboratory fire.) pophysis of a 17 months old gonadectomized male (dba 9 X ced') F1 hybrid WK 431. Mag. X439. FIG. 7.â€”Hypertrophied basophiles; note enlarged negative FIG. 6.â€”Hemorrhagic cysts lined by abnormal basophiles in image of the Golgi apparatus and large nucleoli. No other cell the pituitary of a 23 months old gonadectomized male (ce 9 X types present in this section. From 17 months old gonadecto dbad') F, hybrid WK 615. (Photomicrograph made from lan mized male (dba 9 X ccc?) F1 hybrid WK 431. Mag. X439.

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Fn. 8.â€”Regionof abnormal basophiles showing loss of co FIG. 10.â€”Section of abnormal pituitary showing enlarged hesion of these cells. From WK 431. Mag. X439. sinuses. Section is composed of transitional basophiles and hy pertrophied basophiles. From WK 448. Mag. X93. FIG. 9.â€”Cyst lined by ciliated epithelium at the periphery Fia. 11.â€”Mammary gland of a 20 months old gonadecto of the anterior lobe of the pituitary of an 18 months old gona mized male (dba 9 X ceoâ€•)F1hybrid WK 516. Note secretion dectomized female (ce 9 X dbac3') F1 hybrid WK 448. Mag. globules in the alveolar cells and ducts distended with secre X132. tion. Mag. x 132.

@ months old control female F1 (C,57 Bl@o@Ã§@, of the abnormalities that have been reported were dha aâ€•).' found to be disturbances of chromophobe cells and were observed following treatment with estrogens MICROSCOPIC OBSERVATION OF THE (5, 6, 16). Many spontaneous chromophobe cell MAMMARY GLANDS adenomas have been found and studied in rats of The mammary glands of most of the experi advanced age (37, 40), and following treatment mental animals with hypophyseal tumors were cx with various hormones (36, 38, 39). amined histologically and showed alveolar cells For purposes of differentiating the present series and alveoli filled with secretion and the ducts dis of tumors of the anterior pituitary from chromo tended with secretion (Figs. 11 and 1@). In some phobe adenomas that have been reported, it might of the glands there were areas of hyperplastic nod be said first of all that we are dealing with a differ ules (so-called precancerous lesions) and in four ent endocrine background than that used in the castrate males and a few gonadectomized females, study of chromophobe adenomas. Here the hor mammary gland carcinomas were found. The monal stimulation arises from adrenal cortical hor mothers of these F, animals were dba strain mice. mones in castrate animals. Chromophobe adeno These mice had the milk factor from strain dba, mas have been observed both in intact old mice and therefore with proper endocrine stimulation and rats and following estrogenic treatment in

TABLE 2

OCCURRENCE OF TwoANDCONTROL TYPES OF CYSTS IN THE HYPOPHYSES OF GONADECTOMIZED MIcE5Nuuaaa INTACTF, RECIPROCALHYBRID or CILIAnn CYSTS NoN-cIu&Tan CYSTS TOTALOFALLCYSTS SEX ANIMALS%186 No. % No. % No. 19.88uâ€• 14 7.52 23 12.36 37 14.61Total 178 7 3.94 19 10.67 26 17.309â€˜@&@r 364 21 5.76 42 11.54 63 9.85d' 213 7 3.28 14 6.57 fi 27.02Total 211 14 6.64 48 20.38 57 18.39S9 & aâ€• 424 21 4.95 57 13.44 78

There is little difference in the percentage occurrence between the gonadectomised and control groups, but there iss statistically significant sex difference in the occurrence of non-ciliated CySts in the control F1 hybrid mice.

the mammary tumors might be expected, at least both species. Chromophobe adenomas produce a in intact animals. In other experimental animals condition stimulating hypopituitarism when they where the milk factor was absent, the glands were occur following estrogen treatment, while in this well developed, but hyperplastic areas (so-called series, from the physiological condition of the ani precancerous lesions) and mammary tumors never mal as expressed in the accessory reproductive or occurred. gans, a state of hyperfunction exists. Part of the In studying the mammary glands of castrate hormonal stimulation is directly attributable to animals where no pituitary abnormalities oc the hormones secreted by the adrenal tumors but curred, there was no alveolar development ob the extreme alveolar development of the mam served. The amount of growth up to the stage of mary glands seems to depend upon the presence of alveolar formation of these glands depended on the the abnormal basophiic cells in the pituitary. extent of abnormality occurring in the adrenals. Reasons for believing that these various types of cells found in the abnormal areas of the pituitaries DISCUSSION were basophiles are manifold. Their staining reac The results show that these abnormal pituitaries tion with aniline blue showed that they had a present several significant facts. The first and per great affinity for that dye. The negative image of haps most important, is that in all cases the ad the Golgi apparatus, which is quite different in renal cortical tumors preceded the appearance of normal acidophiles and basophiles, was always like the gross hypophyseal tumors. Second, two types that found in mature basophiles even though at of nodules or tumors were present and both types times it was greatly hypertrophied. This hyper were composed of basophiles, and third, the pitui trophy indicates a heightened activity of the cell tary alterations were correlated with physiological (15). There was obviously great physiological ac changes in the animals that were manifested par tivity from the intense alveolar development of the ticularly in the mammary glands. mammary glands. Severinghaus states that with Very few spontaneous hypophyseal abnormali chromophobe adenomata there is no physiological ties have been reported previously in mice. Most alteration in the animal except that caused by de

struction of the anterior lobe and invasion of ad terrupt the normal contour of the intermedia and jacent tissues. Furthermore, basophile cell dis very occasionally of the posterior lobe. turbances are almost invariably associated with The nodules found in the pituitaries of these changes in the adrenal cortex (30). mice were non-capsulated, but no invasion of other Another factor in favor of classifying the pres lobes or surrounding tissues was observed. Mi ent series of pituitary abnormalities as basophile toses were rare, but evidences of amitotic division were prominent, a condition thought to be non existent in a normal hypophysis. By &tlling this condition amitotic division the authors are merely calling attention to the fact that the multinucleate cells were dividing in this manner or mitoses were incomplete and the cytoplasm did not divide. An other supposition is possible, and that is that mi tosis was occurring too rapidly to permit the cyto plasm to divide. Selye has formed this opinion from studying the pituitaries of rats after hormone treatment (@8). The large transitional basophiles were in orderly acinar groups, while the small transitional cells and the abnormal basophiles were in dense clusters. These small cells may have, as Severinghaus states, @ 1aÂ¼.(@ . passed a phase of hypersecretory activity and have become exhausted. The nodules formed by large @ @@1r1@: @t@t4@@aâ€˜@ , transitional basophiles and small transitional cells follow closely Ewing's description of diffuse hyper plasia in the pituitary in man. @ â€¢@-â€˜!;:..@. â€œOnsections the growth may be diffuse . . . or @ i,.- -,@ f(t @, )â€œ .@( focal adenomas may appear as opaque spots. . . .â€œ @@@ @*.::-@â€˜ : , â€¢@ ,, C â€¢ @@@@ @;â€˜a; .. .. â€¢.â€¢â€¢â€¢â€¢.@@@â€˜.. â€œThearrangement of cells is orderly and mitoses @@@ .@:i-_:Is@.@. -@ â€¢1. @,â€¢,,.. ...â€¢.. . .%:@. are rare. While the gland usually remains solid, small cysts filled with colloid may develop and -I ... hemorrhages may occurâ€• (1@). The relations of cell types and various hormones have been established. Acidophiles are associated with growth-promoting factors; these cells are said to be absent in dwarf mice (33) and there is an overabundance of them in acromegalics, in man (17). Pregnancy affects both acidophiles and basophiles (30). Basophiles are modified by thy Flo. 12.â€”Mammary gland of 18 months old gonadecto roidectomy (31), states of thyroxine deficiency mized female (dba 9 X ccc?') F1 hybrid WK 461. The alveolar (18), and thyroid hyperplasia and adenomata (19). spaces and the ducts are distended with secretion. Mag. X132. Also, evidence has been secured that basophiles

FIG. 13.â€”Mammary gland a(lenoma (at right) and carcino secrete the adreno-corticotropic hormone (@5). No ma (at left) from a 16 1 @â€˜2monthsold gonadectomized male where as yet in the present series have been found (dba 9 X ceoâ€•)F1 hybrid WE 425. In the adenomatous area morphological differences correlated with the pres there is some secretion in the alveoli. Mag. X93. ence of these pituitary tumors other than the unusual development of the mammary glands. disturbances is the size of the tumors. Where From the literature very little reference was found, chromophobe adenonias occur they are described however, to mammary gland growth allied with as large, filling the entire base of the skull and pituitary disfunction. It has usually been modified exerting pressure on the brain (16, 40, et a!.). How by steroid hormone injection. It is known that ever, the tumors described here and those baso mammary development follows treatment with phile adenomas reported elsewhere (17) have never some steroid hormone preparations, and in certain attained this great size. They are discrete nodules strains chromophobe adenomas occur. It is reason not of sufficient voluire to exert great stress on sur able to assume that the development of the mam rounding tissues. They have been observed to in mary glands is due to the interaction of the hor

Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 1949 American Association for Cancer Research. DIcKIE AND W00LLEYâ€”Spontaneous Basophiic Tumors 381 mone with various pituitary factors, or the chro glands are so closely related that the mammary mophobe adenomas are transitional basophile gland development cannot be discussed unless the types and hence would secrete the factors neces role of the pituitary is taken into consideration sary for the mammary gland growth or at least (16). part of its growth. Evidence against the latter is The presence of mammary tumors in certain of the difference in tumor size, and in spontaneous these animals with anterior pituitary tumors and chromophobe adenomas no physiological action on also in the parental strain dba would seem to es the accessory reproductive system has been re tablish for gonadectomized animals the factors corded. That this is true is shown by our observa necessary for mammary tumor production. It is tions that with the presence of the hypophyseal known that in intact animals the proper genetic nodules, the excessive mammary gland develop constitution (such as in strains dba or C3H), hor ment was the constant feature. The mammary monal influences and Mammary Tumor Inciter gland development depends in part on the release (M.T.I.) are necessitiesfor spontaneous mammary of gonadotropic factors (34), a reason for believing tumor appearance generation after generation (3). that the substances associated with the abnormal Now in these animals under discussion, it is pos hasophiles are gonadotropin-like. sible to postulate that the genetic constitution and In parabiotic experiments, where a castrated rat was joined to an intact rat, it has been ob TABLE 3 served that there is an increase in the number of HYPOTHESISTO SHOWTHAT MAMMARYTUMORPRODUC basophiles in the pituitaries of both animals, in TION IN GONADECTOMIZED MALE AND FEMALE MICE terpreted as storage of gonadotropic hormones IS DEPENDENT UPON FouR5 FACTORS (@7, 3,5). Adrenal Hypertrophy of the Golgi bodies, and degranu tumors (br lation occur; these latter indicate an increase in Geneticmonalconsti Pitui Mam secretion and its release (35). Thus it might be as influ tary mary Straintutionenee)dba++Ce M.T.l.t tumors tumors sumed with pituitary basophilism, that : (1) the + + + storage and release of gonadotropins is greatly in +C57 .â€”. + creased. A greater production of gonadotropins Blackâ€”â€”dba9 might affect the mammary glands in these experi. XC57Blackoâ€•++C57Black + Â± Xdbao'++dba9Xceo'++ce9Xdbaoâ€•++ce9XC57Blacko'â€”+C57Black9 + mental mice particularly since the gonads are not + + + present and the hormones from the adrenals may + either not regulate the gonadotropic action of the + 9 Xceoâ€•â€”+ + pituitary as the gonads do, or (@) the adrenal corti * The genetic constitution of the animal, the hormonal influences which cal secretions may be slightly different from those are supplied by the adrenal tumors, the presence of mammary t umor inciter, and the presence of basophilic pituitary tumors. of a normal gonad, or (3) the pituitary secretions t Mammary tumor inciter. may act directly. It is not the purpose of the authors to discuss M.T.I. are still necessary and the hormonal influ the hormonal aspects involved in the present series ences that were supplied by the gonads are now at this time, since the substances elicited from the supplied by the abnormal hypophyses and ad adrenal cortical tumors have not yet been identi renals, to prepare the mammary glands for tumor fled, but to say briefly that the substance or sub development (Table 3). Further, that where the stances elicited from the pituitary and adrenal pituitary tumors are present the mammary glands glands seem to be of a gonadotropic and gonadal are better prepared than with .adrenal-cortical nature respectively. The effects produced on the tumors alone. mammary glands and accessory reproductive or In the F1 reciprocal hybrid mice where the gans are similar to the effects produced by the hor pituitary abnormalities were very infrequent, it is mones from the ovaries or combinations of ovaries evident that the only change other than the pitui and testes when they are present. It is well known tary tumor is the excessive mammary gland devel that there are close relationships between the opment. Even though other animals in these hy hypophysis and gonads and the hypophysis and brid groups had adrenal tumors, the extensive adrenals. Recently much evidence has been re mammary development appeared only when the ported on the interrelation of the pituitary and the pituitary was abnormal. mammary glands through the action of mam The pituitary cell changes in some respects are n@cgenic hormones that affect the duct extension similar to those described by Crooke (7), Severing and lobule alveolar growth (@3, @6).Gardner has hans (30), and Mellgren (@5), in the pituitaries of stated that the hypophysis and the mammary patients with Cushing's syndrome. Basophilia has

been known to occur as a part of this and other sponsible for, and producing, at least one other syndromes. McLetchie (@4) has stated that the tropin, one that affects mammary development castrate condition and Cushing's syndrome have since this seems to be the case in these animals. much in common as in the latter there is frequently Woolley has transplanted an adrenal tumor from a gonadal atrophy. Selye reports that it is very prob cc mouse with a basophilic pituitary tumor and able that in these cases tumorous hypophyses are hyperplastic mammary glands. The transplanted capable of producing an excess of such pituitary adrenal tumor did not cause overdevelopment of hormones as adrenotropic, mammotropic, lacto the mammary glands in the new hosts, cc gonadec genic and perhaps other factors (@9). McLetchie tomized mice (44). It may be assumed that the declares further that: pituitary tumor either directly or indirectly, rather

â€œ. . . it is the rule rather than an exception to than the adrenal tumor itself was responsible for find adreno-cortical hyperplasia associated with the mammary gland growth. Here then, in mice at basophil adenoma and again in some cases of least is a factor that seems to be missing or that adreno-cortical carcinoma, basophilia, that is the has taken another form of expression in man. The relative increase in basophil cells, is present . . present situation then would be analogous to hypersecretory processes of the basophile cells and eosinophilic tumors in man (@) where there are the adrenal cortex are complimentary, the one other factors influenced by the anterior lobe cells. producing the otherâ€• (@4). This would further advance the possibility that In the adreno-genital syndrome, and in inter the pituitary is the primary site of the disturb sexuality, it has been thought that the pituitary ances rather than the adrenals. becomes abnormal first and then the adrenal Castration in rats and mice causes increased changes follow (4). In Addison's disease however, gonadotropic potency. One might assume that the the adrenals become abnormal and the hypophys gonadotropins are transformed into a substance cal changes follow (8) . In this experimental series either identical with adrenocorticotrophic hor the gross adrenal changes are primary. In some mone (ACTH) or similar to ACTH that can act on crosses adrenal cortical tumors occur as early as 8 the adrenals. Other workers have given serious months of age, while the hypophyseal changes are consideration to the idea that since one cell type not grossly observed before 14 to 18 months in the seems to produce so many hormones, the cell types dba X cc crosses and later in other crosses. It is may in effect secrete one or two basic precursors possible to postulate that the pituitary may be (17) that act in various ways according to the thrown into unbalance primarily by castration, cx demands made upon it by other glands. ert its effect on the adrenals in the absence of the It is felt however, that the investigation of these gonads, and the adrenals in turn then become ab tumors is by no means complete. Since there is a normal. The abnormal adrenals then secrete excess gross macroscopic change and histological change, or unusual hormones which react on the pituitaries perhaps there are identifiable biochemical changes and they, then, become abnormal. also. Too, the developmental stages need to be It seems pertinent to further compare the syn studied more completely. drome in mice with Cushing's syndrome in man.4 No cell types have been identified as exact coun This may well be an animal counterpart of Cush terparts of Crooke's change, the hyalinization of ing's syndrome. The latter syndrome is a baffling the basophile cells. It might be speculated that the problem both as to cause and therapy (9). Kepler large and often multinucleate abnormal cells in (@1)in an analysis of the present status of the these mice are similar to the hyaline basophiles or problem has made several assumptions that might Crooke's cells, but the hyalinization in the ha be applied to the mice. He states that the pituitary sophiles of the pituitaries of the mice is less acute, may become overactive primarily and cause the if present at all. These abnormal basophile cells in development of adrenal hyperplasia or neoplasia the mice compare more closely with the hyper and then the abnormal adrenal cortices function trophic amphophiles described by Meligren and excessively. If the abnormal basophilic cells are the others (@,5).Mellgren believes that these cells, the primary cause of the adrenal disturbance, unless amphophiles, are later stages in the degranulation they produced only the adrenotropic agent and no and hyalinization of the basophile cells. This might other tropins, some manifestations should remain be the case in the mouse pituitaries since these following removal of the adrenal tumors. Now, anterior pituitary abnormalities seem to be in well rather than suppose that the basophiles are secret advanced stages of alteration at the time of au ing only one hormone, let us assume they are re topsy. Collection of the early stages was not pos

4 Cushing's syndrome indicates only those cases in which an sible in this series because the experiments were adrenal tumor is present. well advanced at the time the pituitary abnormali

ties were first noted macroscopically. It is hoped gross changes. occurred eventually after the adre that more information on analogies and tM@i1t@vel nals were tumorous. oprnental processes will be learned by a study of pituitaries of younger gonadectomized animals. REFERENCES From the preceding discussion, it is very evident 1. ADDISON,W. H. F. The Cell Changes in the Hypophysis of that while the abnormalities of the pituitaries are the Albino Rat after Castration. J. Comp. Neurol., 28: a fact, their function, i.e. the function of the cells 441â€”463, 1917. 2. BIGGART,J. H. The Hypophysis of the Human Castrate. composing the abnormalities, with respect to the Bull. Johns Hopkins Hosp., 54: 157, 1934. secretions they may produce is very speculative. 3. BITmmt, J. J. 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CORRECTION The title of the last column of Table I in the paper by Bass and Feigelson, Cancer Research, 8: 507, October 1948, should read â€œDry weight meanâ€•instead of â€œWetweight mean.â€•

Margaret M. Dickie and George W. Woolley

Cancer Res 1949;9:372-384.

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