Gonadectomized Mice Research Articles

Autoradiographic localization of dihydrotestosterone binding in the major salivary glands and other androgen-responsive organs of the mouse.

Mouse submandibular glands show an androgen-dependent sexual dimorphism, reflected in higher concentrations in males than in females of bioactive peptides, such as epidermal growth factor (EGF), nerve growth factor, and renin in the cells of the granular convoluted tubules (GCT). Biochemical studies have demonstrated androgen receptors in submandibular gland and other androgen-responsive organs in mouse. We have determined the cellular localization of these receptors using steroid autoradiography. Fifteen adult gonadectomized male mice were injected intravenously with 0.13 microgram or 0.26 microgram [3H]-dihydrotestosterone (SA 135 Ci/mM); some animals were pre-treated with cyclocytidine to stimulate secretion by GCT cells. Animals were killed 15 min, 1, 2, or 3 hr after isotope injection. Steroid autoradiographs were prepared, and some were stained immunocytochemically for EGF. Of the different cell types of submandibular gland, the acinar cells most frequently and intensely concentrated [3H]-DHT; GCT cells also concentrated the hormone, as did a small number of striated duct cells. In the other major salivary glands, the only cells that concentrated the androgen were interlobular striated duct cells in sublingual gland. In prostate, anterior pituitary, and brain a large number of cells concentrated androgen, as has been previously reported. Androgen binding by the GCT cells was a predictable finding, since androgen-induced alterations in composition and form of these cells are well documented. The intense androgen concentration by the acinar cells was an unexpected finding and suggests a hitherto unknown androgen regulation of these cells. An incidental finding was intense concentration of [3H]-DHT in the nuclei of the endothelial cells of the post-capillary venules of the cervical lymph nodes.

Seasonal variation in mammalian striated muscle mass and motoneuron morphology.

Feral white-footed mice are seasonal breeders that undergo predictable cycles of reproductive function. Photoperiod-induced fluctuations in gonadal function of white-footed mice were associated with morphological changes in perineal muscles and their motoneurons. Exposure to short daylengths resulted in testicular regression, decreased perineal muscle mass, and shrinkage of somata and nuclei of motoneurons of the spinal nucleus of the bulbocavernosus (SNB). These effects were reversed by reinstatement of long daylengths. Similar reductions in muscle mass and SNB soma size were seen following gonadectomy of white-footed mice. In addition, dendritic trees of SNB motoneurons were reduced in gonadectomized mice compared with dendritic arbors of intact mice or castrates provided with testosterone capsules. Androgen-mediated annual changes in muscle mass and motoneuron morphology appear to be a natural part of this species' physiology.

Influence of testosterone on hydronephrosis in the inbred mouse strain DDD.

Hereditary hydronephrosis was detected in all of the male mice of DDD inbred strain maintained at the National Institute of Animal Health Japan, but in only a few of the females. From the standpoint that male hormones are related to the development of hydronephrosis in this strain, the incidence and severity of the disease were investigated in gonadectomized mice treated with testosterone. In the males, the incidence of hydronephrosis was 50% (7/14) in the control (gonadectomized) group, and 73.3% (11/15), 100% (13/13) and 100% (12/12) in the 0.1 mg, 3.3 mg and 10 mg treatment groups respectively. The same tendency was observed in the female animals, though the incidence in each group was not so high. In both sexes the degree of severity increased in proportion to the dose of testosterone administrated. Blood testosterone levels were higher in intact DDD mice than in C57BL/6 mice, which had normal kidneys in both sexes. These results suggest that male hormones play a significant role in the development of hydronephrosis.

Nippostrongylus brasiliensis: Effects of testosterone on reproduction and establishment

Gonadectomy of the host reduced the establishment of Nippostrongylus brasiliensis in male mice, but elevated the recovery of worms from female hosts. Similarly, production of eggs by individual females of N. brasiliensis decreased and increased in gonadectomized male and female animals, respectively. Implantation of testosterone-filled capsules caused a dosage-dependent release of eggs by helminths in gonadectomized hosts of both sexes. Maximal production of eggs by nematodes was found when plasma testosterone in the host exceeded 20 ng/ml. Treatment of gonadectomized mice of both sexes with testosterone implants gave recoveries of worms that were similar to the number of helminths in intact male mice. Ligation of the bile duct of male mice that were implanted with testosterone reduced both establishment and reproduction of N. brasiliensis.

Gonadectomy, sex hormones and the growth of tetrathyridial populations of Mesocestoides corti (Cestoda: Cyclophyllidea) in mice

Gonadectomy in mice considerably depressed the volumes of intraperitoneal tetrathyridial populations, orchiectomy being more efficient than ovariectomy. Testosterone propionate accelerated the growth of tetrathyridial populations in gonadectomized mice of both sexes. Oestradiol benzoate was less efficient, but considerably increased the invasion of livers both in intact and in gonadectomized mice. The average size of tetrathyridia was inversely proportional to the size of the populations: tetrathyridia from an orchiectomized mouse were the largest, those from an orchiectomized and testosterone treated mouse, the smallest, and those from an orchiectomized mouse treated with oestradiol, intermediate in size.

Ovarian Influence on Pulmonary Carcinogenesis by Hydrazine Sulfate in BALB/c/Cb/Se Mice<xref ref-type="fn" rid="FN2">2</xref>

Daily administration of hydrazine sulfate, 1.13 mg/mouse for 150 doses, to BALB/c/Cb/Se female mice in various hormonal states varied the incidence and the average number of pulmonary tumors per tumor-bearing mouse as follows: in intact virgins, 90.0% and 3; in breeders, 100% and 14; in gonadectomized mice, 60.0% and 5. Histologically, in intact virgins, 3.3% were carcinomas and 96.6% were adenomas; in breeders, 47.2% were carcinomas and 52.7% were adenomas; in gonadectomized mice, 4% were carcinomas and 96% were adenomas. Many pulmonary carcinomas induced in breeders infiltrated the thoracic wall and the mediastinal organs and metastasized to the adrenal glands and myocardium. The greater biologic and morphologic malignancy of the induced tumors observed in breeders, as compared to those in intact virgin and gonadectomized mice, was associated with an increased ovarian hormone production.

Effect of adrenalectomy and gonadectomy on protein turnover of liver, kidney and spleen of mice.

Half-lives of protein in cell fractions of liver, kidney and spleen from adrenalectomized and gonadectomized mice were determined. The effect of fasting, protein-free and high-protein diet and cortisol administration to adrenalectomized animals was also examined. Decrease in turnover rate of protein was observed in all tissues examined from adrenalectomized and gonadectomized mice, however the degree of decrease was more remarkable in tissues of adrenalectomized mice than in those of gonadectomized mice. Daily administration of a large dose of cortisol accelerated the degradation of labeled proteins. Fasting caused a remarkable retention of protein in liver and kidney but not in spleen and no significant change in turnover rate of proteins was observed in the sham-operated control group, therefore, it was concluded that the changes in half-lives observed in adrenalectomized and gonadectomized mice were mainly attributable to the endocrine deprival and not to the result of change in food intake after the operation.

Development of goitre after methylthiouracil treatment of mice with hypothalamic lesions.

SUMMARY Lesions aimed at the area of the paraventricular hypothalamic nuclei were made by means of stereotaxic electrocoagulation in intact and gonadectomized mice, which were subsequently treated with methylthiouracil. Lesions could be placed with an acceptable degree of precision and with moderate postoperative mortality. Correctly placed lesions made in intact females and castrated males caused a marked inhibition of goitre development, indicating a depressed secretion of thyrotrophin. However, in intact males and spayed females inhibition of goitre formation could not be demonstrated convincingly.

Oxidative enzymes in postcastrational adrenal tumors of mice.

Five oxidative enzymes (succinate, glucose-6-phosphate, isocitrate, β-hydroxybutyrate and 3β-hydroxysteroid dehydrogenases) were demonstrated by histochemical methods in the adrenal glands of 3 strains of mice—DBA /2J, C57B1/6J and A/Jax—3, 6, 9 and 12 months after gonadectomy during development of postcastrational adrenal tumors. Intact animals sacrificed at the same time intervals served as controls. Areas of “A” cell hyperplasia, present in the adrenal cortex of both intact and gonadectomized mice, were characterized by low activity of all 5 enzymes. In contrast, “B” cells, which were present only in the adrenal adenomas of gonadectomized mice, were characterized by high glucose-6-phosphate and isocitrate dehydrogenases, moderate 3β-hydroxysteroid dehydrogenase and low succinate and β-hydroxybutyrate dehydrogenases. This suggested that the “B” cells were metabolically more active than “A” cells. Steroid dehydrogenase was the only enzyme which could be correlated with production of sex hormones by the a...

EFFECT OF GONADAL HORMONES ON SUBMANDIBULAR GLAND AMYLASE ACTIVITY IN MALE AND FEMALE C57BL-6J-6J MICE.

Submandibular gland amylase activity in C57BL/6J mice of both sexes was determined in intact controls, in gonadectomized animals, and in gonadectomized animals following the administration of testosterone propionate or estradiol benzoate. Amylase activity was 3 times higher in the glands of intact males than in those of intact females. Castration caused a reduction in the amylase activity in the glands of males to less than £ the control value, but not to the low level found in females. Activity in the glands of ovariectomized females remained unchanged. The administration of testosterone propionate to gonadectomized males and females raised the activity to the same value in both sexes, a level higher than that observed in intact males. The administration of estradiol benzoate to castrated males raised amylase activity to a value significantly higher than in ovariectomized females receiving the same treatment. An increase in amylase activity does not appear to be dependent upon an increase in submandibular gland weight. These results are discussed in the light of observations reported by previous investigators. (Endocrinology 76: 776, 1965) /CONSIDERABLE interest in the horv>< monal control of submandibular gland activity in mice and other rodents has been shown since Lacassagne (1) demonstrated a sexual dimorphism of this gland in 1940. Raynaud and Rebeyrotte (2, 3) have reported that the amylase activity is higher in the saliva of male mice than in the saliva of female mice and that castration of males lowers amylase activity to female levels. In contrast, Junqueira et al. (4) were unable to detect a difference in amylase activity in extracts of salivary glands from mice of either sex. In an abstract of a paper presented before the American Society of Zoologists in 1957, Harvey (5) reported that following castration there is a reduction in amylase activity in extracts of glands from males, but not in those from glands of ovariectomized Received August 17,1964. This study was supported in part by USPHS Research Grant A-4357 from the National Institutes of Health, by USPHS Research Grant CA-00362-15 from the National Cancer Institute, and by Institutional Grant 83701G from the American Cancer Society. females. She reported further that the administration of either testosterone or estradiol following gonadectomy stimulated an increase in amylase levels in the glands of both sexes. Comparable levels for intact animals of both sexes were not given; nor were any quantitative data presented. Unfortunately, Dr. Harvey died before she was able to prepare a comprehensive account of her work. In the present study, data were obtained from the analysis of submandibular gland homogenates which demonstrate unequivocally that amylase activity in C57BL/6J mice is higher in the glands of males than in those of females, and that testosterone propionate stimulates an increase in activity in the glands of gonadectomized mice of both sexes. In addition, submandibular gland amylase activities found following the administration of graded doses of estradiol benzoate to gonadectomized male and female mice are presented. Materials and Methods One hundred and ten adult C57BL/6J mice, 54 males and 57 females, were used. Eight mice of each sex were reserved as normal, intact con-

Influences of the gonad on the thymus in the mouse

In mice of an inbred Japanese dd-strain, age changes in the thymus weight were studied for both sexes from 5 days until 300 days of age. Further studies were made on effects of gonadectomy on the thymus in both male and female mice which were gonadectomized at 35 days and 100 days of age. 1. The thymus grows markedly at the same rate in both sexes in early life until at about 35 days it reaches a maximum in weight. Then it decreases in weight due to age involution. During adolescence and early maturity until about 100 days the decrease in thymus weight is relatively rapid, and it is more prominent in males than in females. During sexual maturity after 120 days until 300 days so far examined, the decrease in the thymus weight is very slow, and the thymus is significantly heavier in females than in males. 2. The sex difference in the thymus weight is no longer evident in gonadectomized mice. The thymus of the male responds to castration by a remarkable increase in weight, while the thymus of the female is little influenced by ovariectomy. From these considerations it is surmised that the sex difference in the thymus weight is due to that the thymus in the male is suppressed by the testis, namely, the male gonadal hormone.

THE NATURE OF THE SEXUAL DIMORPHISM OF THE SUBMANDIBULAR GLAND OF THE MOUSE1

The weights of the submandibular glands and their water, lipid and nonlipid fractionswere determined in intact, gonadectomized and androgen-treated gonadectomized mice of both sexes. It was found that although the glands are markedly heavier in intact males than in females, the composition of the tissue is the same in both sexes. The loss of weight after gonadectomy is accounted for solely as loss of water in the female, whereas in the male there is loss both of water and dry tissue residue. Treatment with androgen results in the increase in the absolute amount of water and dry tissue residue in both sexes. The present data indicate that the larger size of the glands in intact male and androgentreated mice is due primarily to differential growth of the terminalsecretory ducts.

Effects of estradiol-17-beta-n-valerate on endosteal ossification and linear growth in the mouse femur.

Bone responses were studied in gonadectomized mice treated with amounts of estradiol-17-beta-n-valerate ranging from 0.1 to 20 µg. Linear growth was accelerated more significantly in femurs from orchidectomized mice than those from ovariectomized mice. Small amounts of the estrogen accelerated further linear growth of the femur, while at the same time they promoted the resorption of metaphyseal trabeculae. Larger amounts of estradiol valerate inhibited resorption of metaphyseal trabeculae, and stimulated endosteal osteogenesis. Some of the possible mechanisms underlying these observations are discussed.

The influence of sex hormone status on the susceptibility of mice to chloroform-induced necrosis of the renal tubules.

SUMMARY 1. Adult male mice of two strains developed extensive necrosis of the renal tubules after exposure to low concentrations of chloroform vapour. Adult females showed no renal damage after equivalent exposures. 2. Adult females became fully susceptible to necrosis after treatment with androgens. The susceptibility of males was greatly reduced by treatment with oestrogens. 3. Castration removed the susceptibility of the males in one strain but did not completely remove it in another. The residual susceptibility of castrates was abolished by adrenalectomy. 4. Male mice under 11 days old were insusceptible to necrosis even after massive doses of androgen. Between 11 and 30 days they were susceptible if given androgen. Thereafter they became spontaneously susceptible. 5. Liver damage occurred in nearly all exposed mice and was not correlated with sex hormone status. 6. Susceptibility could be induced in gonadectomized mice by methyl testosterone, testosterone propionate, dehydroepiandrosterone and progesterone, and by large doses of cortisone acetate. 7. Preliminary exposure to carbon tetrachloride completely protected susceptible males against renal damage by chloroform.

OVARIAN TUMOURS IN MICE IN PARABIOTIC UNION

Spontaneous ovarian tumours in mice are extremely rare. The incidental finding of ovarian tumours during Furth & Butterworth’s (1936) attempt at the experimental production of leukaemic changes by means of X-rays was a fortunate coincidence, which has increased our knowledge of these tumours. The study of ovarian tumours produced in mice by roentgen irradiation has since been continued by Furth & Boon (1947), Furth (1949) and Bali & Furth (1949). Ovarian tumours following X-ray treatment in mice was confirmed by several authors s. a. (Geist et al. (1939), Kaplan (1947), Lick et al. (1949), Gardner (1950), Kaplan (1950) and Mühlbock (1951). More recently, ovarian tumours were observed in non-irradiated ovaries grafted into the splenic or pancreatic tissue of gonadectomized rats and mice (Biskind et al., 1944, Furth et al., 1947, Li et al., 1947 a, Biskind et al., 1948, Li et al., 1949, and Lankner et al., 1950). The pathogenic mechanism of these tumours is a matter of considerable interest. There is evidence that the development of tumours in intra-splenic grafts is the result of a hormonal imbalance (Biskind et al., 1944, Li et al., 1947 b, Gardner, 1948, and Li et al., 1949). This hormonal imbalance consists in a relative overproduction of gonadotrophic hormone by the hypophysis so that the exactly adjusted hormonal relationship between the pituitary and the ovarian hormones is changed. This prolonged over-production of gonadotrophic hormone leads to the development of ovarian tumours.

PHYSIOLOGICAL FACTORS AFFECTING THE RESPONSE OF EXPERIMENTAL WOUND HEALING TO ACTH

The formation of experimentally-produced granulation tissue can be inhibited by ACTH. It has been shown that pitressin tannate and chorionic gonadotrophin do not have this effect. No gonadotrophic activity could be detected in the sample of ACTH used. Hypophysectomized mice became progressively and rapidly less responsive to ACTH. The gonads were found to be essential for inhibition to occur. Adrenalectomized mice at the beginning of the breeding season, adrenalectomized pregnant mice, and adrenalectomized mice pre-treated with chorionic gonadotrophin showed inhibition of healing by ACTH. Adrenalectomized mice pre-treated with follicle-stimulating hormone or pregnant mares' serum gonadotrophin were not inhibited. Cortisone acetate causes inhibition in intact and gonadectomized mice. Neither testosterone, oestriol, nor oestradiol had any effect; but progesterone gave definite impairment under the conditions of the experiment. It is suggested that the gonads are capable of responding to ACTH under the influence of luteinizing hormone.

Development of adrenal cortical adenomas in ovariectomized mice injected with physiologic doses of sex hormone.

Since the development of adrenal cortical adenomas in gonadectomized mice has been inhibited by the implantation of pellets of estrogenic hormone, it might be inferred that withdrawal of gonadal secretion is of primary importance in precipitating the development of the castration-induced adenoma of the mouse adrenal cortex. The objective of the experiments being reported was to determine the effect of the administration of regulated, “physiologic” amounts of estrogenic or androgenic hormone on the induction of adrenal cortical tumors in susceptible ovariectomized mice. Mice of the NH strain were used in these experiments. NH females develop adrenal cortical tumors spontaneously following ovarian regression, and precociously if gonadectomized. Adrenal cortical adenomas are present in most NH mice of both sexes 5 months after removal of the gonads. Four NH males and one female whihc had been gonadectomized at 23 days of age received (by subcutaneous injection beginning 2 weeks after operation) 0.05 microgram of estradiol dipropionate in sesame oil at intervals of from 5 to 9 days. This treatment with estrogenic hormone provided for constant estrus, as determined by vaginal smear. These 5 animals were autopsied 6 months after the institution of hormone treatment. Administration of hormone was suspended 53 days before autopsy in 2 NH females which received similar treatment. Two ovariectomized NH females received the same treatment with the estrogen and in addition were injected weekly with 150 μg of progesterone in oil. Two additional ovariectomized females received the estrogen plus progesterone, but treatment was suspended 47 days before autopsy. Two NH females ovariectomized at 21 days of age were given 17.5 μg of testosterone propionate in oil weekly by subcutaneous injection beginning 2 weeks after operation. This treatment was continued until the animals were autopsied 5 1/2 months after ovariectomy. Three mice were similarly treated except that they received no injections of hormone for the 46 days preceding autopsy.

Neoplastic Transformation of Granulosa Cells in Grafts of Normal Ovaries into Spleens of Gonadectomized Mice&lt;xref ref-type="fn" rid="fn1"&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/xref&gt;

Neoplastic Transformation of Granulosa Cells in Grafts of Normal Ovaries into Spleens of Gonadectomized Mice&lt;xref ref-type="fn" rid="fn1"&gt;&lt;sup&gt;1&lt;/sup&gt;&lt;/xref&gt;