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1804.Full.Pdf Formation of Adenomata in Hypophyses of Rats Subjected to both Subtotal Thyroidectomy and Administration of'311, and its Prevention by Feeding of Desiccated Thyroid1 w. E. GRIESBACH,2 J@ L. CHAIKOFF, C. W. NICHOLS, JR., AND ROBERT C. GOLDBERG@ Department of Physiology, University of California, Berkeley, California SUMMARY The tumorigenic effect of subtotal thyroidectomy on the pituitary was studied in 212 Long-Evans female rats. Of the 133 pituitaries that contained adenomata after 2 years, 76 % were of the thyrotropic cell type. This finding confirms earlier work on the effect of long-term thyroxine deficiency. The feeding of desiccated thyroid reduced the @)ercentages of thyrotropic cell adeno mata from a mean of 60 to a mean of 33. Thyroid feeding reduced the percentages of all adenomata from a mean of 72 to a mean of 53. A single administration of 1 @ecof 1311did not increase the number of, or tendency towards, anaplastic changes in @)ituitary adenomata of subtotally thyroidectomized rats fed a stock diet. The same dose of 1311did not increase the incidence of pituitary adenomata iii unoperated animals fed either the stock diet or the stock diet supple mented with desiccated thyroid. Two rats that developed malignant thyroid car cinomata after receiving 1 @.icof131Jdid not show cell changes or adenomat.a in the pi tuitary. The absence of cytologic signs of thyroxine deficiency in the pituitary cells of these rats supports the view that irradiation per se might well have been the cause of the neoplasms found in their thyroid glands. The pituitary adenomata found in 124 unoperated rats, treated in the same way as the subtotally thyroidectomized groups with respect to ‘@‘Idosageand thyroid feed ing, closely resembled those found in the normal control group in number as well as in cell type. It has been knowii for almost 2 decades that hyperpla.sia study of the TSH-producing cells in the pituitary, lend as well as benign and nialignant neopla.sms develop ill support to this interpretation. The literature dealing thyroid glands of Irlice and rats in which, for prolonged with this subject has been reviewed by Bielschowsky (2) periods, a state of thyroxine deficiency has been estab and Bielschowsky and Horning (3). lished by any one of the following means : the feeding of a A matter of some interest as well as practical importance low-iodine diet (1, 2) ; the administration of anti-thyroid arose in the case of experiments in which irradiation was substances (5, 15, 25) ; irradiation of the gland by 131J, applied to the rodents' thyroid gland, either alone or in astatine (211At), or X-rays (7, 12, 14, 19) ; and subtotal combination with chemical or surgical means, for inducing thyroidectomy (6, 14). There is good evidence for the the thyroxine deficiency. In most cases, especially in the belief that the neopla.st.ic changes should be attributed to early work with large doses of 131J(up to 875 j@c),the irra the hyperplasia of thyrotropic hormone (TSH4)-producing diation had been sufficiently strong to destroy much or all cells in the anterior pituitary glands of the animals suffer of the thyroid tissue (11, 12). In later experiments the ing from long-terni thyroxine deficiency. Assays of TSH 131J was given in a considerably reduced dose (25—40 Mc). contetits of the hypophysis and blood, as well as cytologic It was shown that hereby the number of experimentally induced malignant thyroid neoplasms was definitely higher 1 This work was supported by a contract from the United States Atomic Energy Commission (UCB-34P11-3). (22). It could further be seen that hyperplasia of the 2 Department of Endocrinology, Medical Research Council, thyroid epithelium surrounding the neoplasms was a con Dunedin, New Zealand. stant feature of these glands. It was assumed that this 3 Department of Medicine, Permanente Medical Group and was significant for the strong and permanent action of Kaiser Foundation Hospital, Oakland, California. thyrotropic hormone. No signs of radiation damage were 4 The following abbreviations are used: TSH, thyrotropic hormone; PAS, periodic acid-Schiff; PTA, phosphotungstic acid; found. The question whether ilradiation per se can induce AF, aldehyde fuchsin. thyroid tumors was discussed in another study in which Received for publication March 4, 1965; revised June 7, 1965. either both lobes or a single lobe of the thyroid gland had 1804 Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 1965 American Association for Cancer Research. GRIESBACH et al.—Hypophyseal Adenoma after Thyroidectomy and 131J 1805 TABLE 1 ADENOMATA IN PITUITARIES OF INTACT FEMALE RATS PvruITARIEsGROUPTREATMENTNo. ADRNOMATATotal CLASSIFICATIONOP OF RATS ADENOMATACYTOLOGIC EXAMINED (76%)INoneVI1 47WITH 21% 45Thyrotrophs3 (14%)Gonadotrophs2Chromophobes16 (60%)VIIDesiccated@c‘@‘Iinjected3410293 (30%)16 (70%)VIII1 thyroid in diet5723403 (13%)416 (82%)Total17171429@c1311plus desiccated thyroid in diet3317510314 (13%)10 (14%)52 (73%) TABLE 2 ADENOMATA IN PITUITARIES OF SUBTOTALLY THYROIDECTOMIZED FEMALE RATS PITUITARIES WITH CYTOLOGIC CLASSIFICATION OF ADENOMATA No. OF RATS ADENOMATA No. or Gaow' TREATMENT ADENOMATA r'ANAPLASTIC EXAMINED CHANGES IN: CLASSIFIED Total % Thyrotrophs Gonadotrophs Chromophobes II Subtotal thyroidectomy only 67 42 63 36 (86%) 3 6 (14%) 45 8 (18%) III Subtotal thyroidectomy plus 49 40 82 33 (83%) 4 5 (13%) 42 10 (24%) 1 pc 1311injected IV Subtotal thyroidectomy, 1 @IC 60 32 53 22 (69%) 3 8 (25%) 33 5 (15%) 1311 injected plus desiccated thyroid fed V Subtotal thyroidectomy plus 36 19 53 10 (53%) 2 7 (37%) 19 2 (10%) desiccated thyroid fed Total 212 133 63 101 (76%) 12 (9%) 26 (20%) 139 been irradiated (21). There, most of the carcinomata 1 and 2) received by i.p. injection 1 j@tCofcarrier-free 131J were found in rats in which both lobes had been irradiated, in 0.1 ml of normal saline. Twenty-four hr later the but 2 carcinomata also appeared in singly-irradiated lobes isthmus contained about 2 % of the radioactivity. and 1 in an opposite, shielded lobe. Hence, it could not One week after either the subtotal thyroidectomy or be decided whether thyroid carcinogenesis resulted from 1311 injection, feeding of the thyroid-supplemented diet prolonged TSH stimulation or was a direct consequence of was begun. The diet was prepared by adding desiccated the irradiation. In a more recent paper, Goldberg et al. thyroid powder (Lilly Thyroid, U.S.P., No. 58) to the (14) reported finding a papillary and a follicular carcinoma stock diet to yield a concentration of 250 nig/kg. It was in intact thyroid glands of rats that received but a single shown in a separate experiment that the uptake of ‘@‘Iby /hc of 131J The present report deals with the 383 pitui the thyroid glands of the rats fed this diet was almost com taries excised from the rats whose thyroids had been de pletely suppressed. scribed in this latter study (14). In these pituitaries, The rats were maintained for 2 years, and the survivors many adenomata were found; they are described in detail, were killed with ether. Those that failed to survive for particularly with regard to the hormonal conditions that the 2 years usually died of chronic respiratory disease, may have influenced their development. and autopsies were not performed on all of these animals. The thyroid glands of the rats of Groups I and VI-Vill MATERIALS AND METHODS were excised and weighed. In all other rats, the tracheas A total of 876 female, Long-Evans rats were randomly along with the paratracheal tissues were removed en bloc selected at 5-6 weeks of age, divided into 8 groups, and and fixed in 10 % neutral formalin. The pituitary glands treated as shown in Tables 1 and 2. After weaning, all were removed and fixed in sublimate-formol (9 : 1, v/v). rats were maintained on a nutritionally adequate diet These glands were sectioned at 2.5 and 5 @,and sections (Diablo Double-Check Labration) containing 3 @sgof from each were mounted on 4 slides, with equal distribution iodine/gm5 (20). onto each slide of the material cut at various depths of the Subtotal thyroidectomy was performed under light block. They were treated as follows : Slides 1 and 2 with ether anesthesia. Both lateral lobes of the thyroid were periodic acid-Schiff (PAS) stain and PAS stain in combi removed, leaving only the isthmus, which weighed about nation with Orange G-phosphotungstic acid (PTA), re 0. 1 mg. One week later, each rat of certain groups (Tables spectively ; Slide 3 with aldehyde fuchsin (AF) usually 5 We are indebted to Dr. G. La Roche for this determination. followed by Orange G-PTA, without previous oxidation Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 1965 American Association for Cancer Research. 1806 Cancer Research Vol. 25, November 1965 TABLE 3 SUMMARY OF CYTOLOGIC FINDINGS IN THE PITUITARIES OF RATS WITH THYROID CARCINOMA GroupTreatmentpituitaryIISubtotal of ratsTyreoftlurroidType of pituitary neoplasmClassification of cells in thyroidectomyPapillaryNone3 cysts in chromophobic areas; few TSIF' foundIISubtotal and LH cells; acidophils not thyroidectomyPapillary(a) Large adenoma; (b) small TSH and “T―-granulated cells; few anaplastic adenoma(a) acidophils; (b) atypical cells, probably basophilsIVSubtotal thyroidectomy + 1 pc cells; LH cells and groups of presentVIInjection1311 injection + addition of acidophils desiccated thyroid to dietPapillaryNone“T―-granulated @ of 1 1311FollicularNoneTSH cells, LII cells, acidophils present in quantitiesVIIIInjection normal of 1 sc 13h1+ addition cells, LH cells present; acidophils in of desiccated thyroid to dietPapillaryNoneTSH creased in numbers a TSH, thyrotropic hormone; LH, luteinizing hormone. with permanganate ; and Slide 4 with the rapid Mallory The number of gonadotroph adenomata was not changed stain of Churg and Prado (4). The last is well suited for by thyroid feeding, but that of chromophobe adenomata counterstaining AF-stained tissues.
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