Anterior Pituitary Powder Research Articles

Loss of mitotic response of the thyroid gland to TSH in hypophysectomized rats and its partial restoration by anterior and posterior pituitary hormones.

AbstractThe follicular cells of thyroid glands of intact rats respond to TSH challenge by an intense mitotic stimulation and a higher functional activity as measured by the epithelium height. In hypophysectomized rats the same intense response was observed when the thyroid was challenged immediately after hypophysectomy. However, one week later the mitotic response was almost completely absent and the functional response much weaker. This indicates that, although TSH is the main regulating factor of the functional activity and cell proliferation of the thyroid follicle, its effect is conditioned by additional factors. These factors may originate both from the anterior and/or posterior pituitary, since a significantly higher response was obtained by injecting anterior or posterior pituitary powder during the interval between hypophysectomy and TSH challenge. Prolactin, vasopressin and oxytocin also boosted the mitotic response to TSH. Posterior pituitary powder alone (without TSH challenge) was not mitogenic for the thyroid gland either in intact or hypophysectomized rats, while it was mitogenic for the adrenal glomerulosa. The low thyroid mitotic responsiveness does not reflect a general unspecific effect on mitoses of all cells, since the esophageal epithelium was not similarly affected by hypophysectomy or by replacement therapy.

Detection of Serum Vitellin in Hepatic Tissue of Chicks by the Use of Fluorescent Antibody Technique

Labelling antibodies with fluorescent dyes have been used to determine histological localizations of antigens in tissues of various viscera by many workers since COONS et. al. (1942) established a fluorescent antibody technique.This investigation was performed to study the mechanism of vitellin production in the hepatic tissue of chickens using the fluorescent antibody technique. An Indirect layer method was employed: this method differs from the direct method in which specific antibody (γ-globulin) is first attached to the antigen in the cell as a layer and then fluorescent anti-globulin (antibody of the layered intermediate antibody) is attached to it to detect the site of antigen. This technique has been conveniently used especially in cases where either pure form of antigen (such as serum vitellin) or high titer of desired antibody is difficult to obtain.Authors used the layering method using rabbit anti-laying hen serum (saturated with cock serum in order to remove the species-specific antibody) as an intermediate and fluorescent chick antirabbit-γ-globulin serum as a demonstrator.Anti-rabbit-γ-globulin chicken serum was conjugated with fluorescein isothiocyanate (F.I.T.C.).Tissue blocks of chicken liver were fixed in acrolein buffered solution (acrolein 10%, 0.01M phosphate buffer (pH 7.2) 90%), and then frozen rapidly on crushed dry ice. Frozen sections were cut at 7 to 10μ with a microtome in a cryostat.The tissue sections were first attached to the rabbit anti-laying hen serum and then fluorescein antibody-conjugates.Observations were made with an Olympus fluorescence microscope using a high-presure mercury-vapour lamp as a source of ultraviolet light.Results were as follows:1. Hepatic tissues of the laying hens and the immature hens treated with C.A.P. (chicken anterior pituitary powder) emitted distinctly yellow-green fluorescence when exposed to the ultra-violet light, but not the tissues of male chicks.2. Antigen (vitellin) was detected in the cytoplasm, but not in the nucleus.3. The fluorescence intensity in the hepatic cells seemed to be corresponding with the intensity of the serum vitellin reactions.

Influence of the anterior pituitary on the aldosterone secretory response to dietary sodium restriction in the rat.

SUMMARY Studies of the rate of aldosterone production in vitro of adrenals of rats hypophysectomized before dietary sodium restriction showed that hypophysectomy not only prevented the increases in aldosterone production observed in intact, Na-deprived rats, but also depressed the level of aldosterone production to below that of intact rats maintained on a normal diet. Rats hypophysectomized for a similar period of time but maintained on the normal diet showed a similar decrease. Experiments on adeno- and neuro-hypophysectomized rats indicated that the pituitary factor required for the normal mineralocorticoid response to dietary sodium restriction resides in the anterior pituitary. Treatment of hypophysectomized rats during dietary sodium restriction with doses of a long-acting corticotrophin (ACTH) prevented adrenal atrophy and maintained a normal glucocorticoid response to intravenous injections of ACTH, but failed to increase aldosterone production rates in vitro to levels above that of intact rats on a normal diet; it also failed to restore the enhanced adrenocortical sensitivity to the stimulating effect of aldosterone production of intravenously injected ACTH which is characteristic of acutely hypophysectomized, Na-deficient rats. Treatment with anterior pituitary powder (8–12 mg./day) for similar periods, however, restored the aldosterone production of adrenals in vitro of hypophysectomized, Na-deprived rats to levels nearly indistinguishable from those of acutely hypophysectomized, Na-deprived controls. The same doses of anterior pituitary powder were shown not to have any demonstrable effect on the aldosterone production of adrenals in vitro of intact rats on a normal diet. These results are interpreted as indicating the existence of a pituitary factor other than ACTH which stimulates aldosterone secretion. This factor does not appear to act directly on the adrenal cortex or to stimulate the secretion of specific glomerulotropic substances, but probably exerts its effect by maintaining the normal functional capacity of some as yet undefined tissues which secrete glomerulotropic substances in response to dietary sodium restriction.

Somatotropin as the non-ACTH factor of anterior pituitary origin for the maintenance of enhanced aldosterone secretory responsiveness of dietary sodium restriction in chronically hypophysectomized rats

Somatotropin treatment in chronically hypophysectomized, sodium-deprived rats effectively restored to treated animals the distinct and enhanced aldosterone secretory responsiveness of the adrenal which characterizes the adrenals of intact rats subjected to dietary sodium restriction, but absent in chronic and nontreated, adenohypophysectomized or totally hypophysectomized rats subjected to similar conditions of dietary sodium restriction. Treatment with β 1−24ACTH(Zn) alone was ineffective, albeit adrenal weight and glucocorticoid secretory responsiveness were effectively maintained. The observed efficacious effect of somatotropin is similar and indistinguishable from the previously demonstrated effect produced by treatment of anterior pituitary powder in chronically hypophysectomized, sodium-deprived rats. The earlier findings that somatotropin is without any direct or specific stimulatory effect on aldosterone secretion and that treatment of intact, sodium-repleted rats with anterior pituitary powder is devoid of any adrenoglomerulotropic influence on the observed refractoriness of the aldosterone secretory response to stimulation justify the inference that somatotropin is the relevant non-ACTH anterior pituitary factor imperative for the presumed normal maintenance of agencies which mediate the characteristic adrenocortical changes in aldosterone secretory capacity in intact, sodium-deprived rats.

Immunological and biological cross-reactivity of chicken and mammalian gonadotrophins.

Rabbit anti-LH-S5 plasma was tested in the ovarian ascorbic acid-depletion (OAAD) assay against NIH-LH-B1, sheep anterior pituitary powder (SAP), chicken anterior pituitary powder (CAP) and chicken plasma. The anti-ovine LH destroyed the OAAD activity of LH-B1 and SAP but did not significantly affect CAP or chicken plasma. Agar immunodiffusion showed precipitin bands between theanti-ovine LH and LH-S5, LH-B1, porcine LH and SAP but not normal rabbit serum or CAP. When guinea pig anti-CAP plasma was allowed to diffuse on agar, bands formed with CAP and chicken plasma but not with LH-Bl or SAP. These results confirm and extend previous indications of a species difference between chicken and mammalian gonadotrophin. (Endocrinology 80: 1182, 1967)

Maintenance and function of corpora lutea in rabbits depend on estrogen.

Pregnant rabbits with one X-irradiated ovary which contained normal-appearing corpora lutea (CL) and interstitial tissue aborted after removal of the contralateral normal ovary containing all ovarian components. The suspicion that estrogen may be essential for CL maintenance and function was confirmed by the injection of estradiol (2–4 μg/day) into rabbits with only an X-irradiated ovary containing just CL and interstitial tissue. Pregnancy was maintained in these females and both the CL and the ovarian effluent blood contained chemically measurable quantities of progesterone. Withdrawal of estrogen caused cessation of progesterone synthesis and secretion and led to abortion. At doses used, neither purified NIH-LH nor crude anterior pituitary powder could, in the absence of estrogen, maintain either the morphology of corpora lutea, or the synthesis of progesterone, or pregnancy. The conclusion is reached and documented that estrogen may act directly on the CL but whether its action is only luteotrophic, or both luteotrophic and steroidogenic, remains to be determined. (Endocrinology80: 938, 1967)

ENDOCRINE CONTROL OF THE ADRENAL IN CHICKENS.

The sulfuric acid-induced fluorescence of corticosterone in the adrenal venous plasma of immature White Leghorn cockerels was studied following hypophysectomy, autotransplantation of the pituitary to the kidney, and following accidental or deliberate damage to the median eminence. That the fluorescence technique measures corticosterone in chicken plasma is supported by observations that the concentration of fluorescing material in the adrenal venous plasma of the chicken (a) was twice that found in the peripheral plasma, (b) increased following injection of ACTH or crude chicken anterior pituitary powder, (c) decreased after hypophysectomy, and (d) had the greatest intensity in that area of a thin-layer chroma togram which had the mobility of standard corticosterone. Significant decreases were observed in adrenal weights and in the corticosterone concentration in the adrenal venous plasma of hypophysectomized birds, and birds bearing pituitary autografts, when compared to sham-operated controls. These data indicate that the chicken adrenal is dependent upon a trophic substance for its regulation. In spite of near-normal histological appearance, the transplanted pituitary cannot secrete ACTH when removed from the sella turcica. After pituitary removal, the fluorescent material found in the adrenal venous plasma of the chicken remains high. These data indicate either that a nonspecific substance in the adrenal venous plasma of the chicken contributes this fluorescence or that the chicken adrenal is capable of producing corticosterone after the adenohypophysis is removed. The latter possibility is supported by the finding that damage to the median eminence significantly decreased the concentration of this fluorescent material when compared to the concentration observed in the plasma of hypophysectomized birds. (Endocrinology75: 192, 1964)

Effects of hormone administration on milk production of underfed rats.

Attempts were made to determine whether the reduction in milk output induced by underfeeding could be overcome by administration of hormones. Half-fed female rats were injected daily with one of the following preparations, from the 4th–17th days postpartum: 15 or 25 mg anterior pituitary powder; 1 mg growth hormone; 10 µg thyroxin/100 g body wt.; 0.5 or 1 mg cortisol acetate; 1 mg prolactin; 1 IU oxytocin; and the latter three hormones in combination. None of the hormones significantly increased litter weight gains, although cortisol acetate, prolactin, and oxytocin each produced slight gains during the later part of the lactation period. These results indicate that the decrease in milk production associated with underfeeding is due primarily to nutritional inadequacy and not to reduced endocrine function. It is believed that the suckling stimulus maintains the secretion of the hormones essential for milk production and removal at optimal or near optimal levels, even in the absence of adequate food intake.

Follicular growth and ovulation in hypophysectomized hens.

Growth of ovarian follicles was induced in hypophysectomized laying hens by daily intramuscular injection of mammalian FSH, PMS or chicken anterior pituitary powder (CAP). The mammalian gonadotropins caused a pronounced increase in the number of follicles entering the initial phase of yellow yolk deposition, but did not support growth of follicles to mature size. The disproportional accumulation of small follicles resulted in the complete obliteration of the normal follicular hierarchy. CAP produced no ovcrstimulation of the smaller follicles and; in a significant number of birds, established a follicular order similar to that found in intact hens. In some CAP treated hens follicles developed to mature size by the end of the eight day experimental period. With a timed sequence of injection of ovulation-inducing substances up to five consecutive daily ovulations were obtained. The number of ovulations was limited by the ability of follicles to attain ovulable size and was independent of the type of gonadot...

In vitro synthesis of C14-corticotropin by isolated rat anterior pituitary: effect of adrenalectomy.

A method is described for the measurement of C14-corticotropin synthesis by isolated rat anterior pituitary tissue. Intact pituitary glands were incubated with C14-phenylalanine; carrier anterior pituitary powder (porcine) was added and corticotropin extracted with hot glacial acetic acid and purified with oxycellulose. The oxycellulose concentrate contained corticotropin (as measured by its ability to cause enlargement of the adrenal gland of hypophysectomized rats) and was free of C14-phenylalanine; on dialysis the radioactivity in the oxycellulose concentrate behaved like purified corticotropin. The rate of C14-corticotropin synthesis by isolated rat anterior pituitary was linear for 4 hr and was increased in pituitaries from adrenalectomized rats. Adrenalectomy did not increase amino acid incorporation into total protein of anterior pituitary; the effect of adrenalectomy was only apparent when pituitary protein was fractionated and oxycellulose concentrates prepared, which is for a specific biological effect of adrenalectomy on corticotropin synthesis.

Mammary lobulo-alveolar growth induced by anterior pituitary hormones in adrenoovariectomized and adreno-ovariectomized-hypophysectomized rats.

Summary1. Mature virgin female Car-worth rats of the CFN strain were adreno-ovariectomized and were then injected thrice daily for 10 days with saline, 2 mg bovine STH, 30 I.U. ovine prolactin, 20 mg bovine anterior pituitary powder, or with a combination of 1.33 mg bovine STH and 20 I.U. ovine prolactin. Treatment with STH and prolactin together or with anterior pituitary powder induced moderate to extensive mammary duct and lobulo-alveolar growth, while treatment with prolactin and STH alone induced mainly duct and end bud development. 2. Two groups of rats were hypophysectomized and 3 days later were adreno-ovariectomized. They were then injected thrice daily for 10 days with saline or with a combination of 1.33 mg STH and 20 I.U. prolactin for 10 days. The latter treatment induced extensive duct and moderate lobulo-alveolar growth. Completeness of ovariectomy, adrenalectomy and hypophysectomy was checked in all rats upon autopsy. Sixteen additional adreno-ovariectomized rats which were given tap water...

Concentration of corticotrophin by percolation.

Percolation of commercial anterior pituitary powder with decreasing concentrations of ethanol (95 → 70%) and increasing concentrations of acetic acid (0 → 0.5 M) yielded a peak of corticotrophin activity with 70% ethanol containing 0.5 M acetic acid. The material assayed at about 14 units/mg peptide. Further purification by curtain electrophoresis yielded a preparation with 21 units/mg peptide.

Studies of pathways of glucose metabolism of endocrine tissues.

Pathways of glucose oxidation using glucose-1-C14 and glucose-6-C14 have been studied in vitro in adrenal, ovary, testis, pituitary and parathyroid slices. The results indicate the existence of an active hexose monophosphate pathway in all endocrine glands studied. Although in similar experiments TSH in vitro was capable of stimulating glucose oxidation in thyroid (1), no effects of FSH, LH, chorionic gonadotrophin or anterior pituitary powder were observed in ovary or testis. Neither ACTH nor 3′5′ cyclic adenosine monophosphate inhibited glucose oxidation in slices of adrenal cortex. Possible causes for this absence of inhibition are discussed in light of the recent work by Haynes on the mechanism of action of ACTH (6). The hexose monophosphatc pathway for glucose metabolism in endocrine tissue is a source of TPNH, and this cofactor may be of importance in the regulation of hormone production.

The production of hypertension, nephrosclerosis and cardiac lesions by methylandrostenediol treatment in the rat.

The “hyalinosis syndrome” is the name applied, by Selye (1946, 1950), to the cardiovascular-renal changes which occur in salt-treated, unilaterally nephrectomized rats administered such steroids as desoxycorticosterone acetate (DCA) or Reichstein’s Compound S (desoxocortisone) for prolonged periods. The characteristic changes of this syndrome are hypertension, renal and cardiac hypertrophy, nephrosclerosis, glomerular hyalinization, “rheumatic-like” cardiac nodules and frequently periarteritis nodosa. Up to the present time such changes have been observed only after treatment with steroids known to possess appreciable mineralocorticoid activity. This fact, plus the observation that certain stresses, as well as the administration of lyophilized anterior pituitary powder (LAP) or alkaline pituitary extract (APE) also produced the hyalinosis syndrome, led Selye to postulate (1946, 1950) that similar pathological conditions in the human might be pathogenetically related to hyperfunction of the anterior pituitary-adrenocortical system involving the increased secretion of mineralo-corticoids. Selye and his co-workers have further observed that experimental nephrosclerosis and hypertension were more readily produced in female or castrate male rats than in intact males

Relation between adrenal and pituitary glands and urinary excretion of neutral reducing lipids in the rat.

The neutral reducing lipids were determined in the urine of rats. Adrenalectomy decreased the excretion of this material to one-half the control value. Neither ACTH, growth hormone, nor anterior pituitary powder significantly increased NRL excretion. Posterior pituitary powder increased the excretion approximately 3-fold. This response is considerably diminished in adrenalectomized rats.

Differentiation of growth hormone from the pituitary factor which produces diabetes.

SummaryGrowth hormone was prepared from a glacial acetic acid extract of anterior pituitary powder. The product was equal in growth-promoting activity to other purified preparations but did not produce diabetes when administered in relatively large doses to dogs, indicating of the pituitary hormone that the diabetogenic factor is distinct from the growth

Quantitative Effects of Implantation of Cattle Anterior Pituitary Powder on Gonads of Immature Rat

Summary1. A method is described for administration of pituitary powder by the tablet implantation technic. 2. Statistically significant responses were obtained with 50 mg of powder in case of testes, uteri, and ovaries of 22- to 24-day-old white rats. 3. These responses were measured by organ weight gains rather than by histological changes or vaginal smears.

Extracts of Anterior Pituitary Growth Hormone

SummaryPhosphate buffer extracts of growth hormone were highly active when assayed in terms of anterior pituitary powder. This method of extraction permitted a careful control of pH and gave high yields of hormone. These extracts had less tendency to form precipitates than those prepared with sodium hydroxide. The addition of urea retarded such precipitation.

Exophthalmos in Thyroidectomized Guinea Pigs by Thyrotropic Substance of Anterior Pituitary, and the Mechanism Involved

We pointed out1 that chronic, progressive, bilateral exophthalmos, produced in immature (2-3 months old) rabbits, maintained on a diet of alfalfa hay and oats, by the daily intramuscular injection of 0.05-0.1 cc. methyl cyanide, occurs in association with thyroid insufficiency (that is, (a) marked hyperplasia of the thyroid or (b) thyroidectomy). It is obvious, therefore, that exophthalmos is not dependent upon either a normal or abnormal thyroid secretion. Neither do the medulla and cortex of the suprarenal gland have any specific or initial relation to exophthalmos.To obtain further information on the nature of exophthalmos we have produced it in young normal and thyroidectomized guinea pigs by using acetic acid extracts of anterior pituitary powder (Armour) that Spaul,2 Uhlenhuth and Schwartzbach,3 and Loeb and Bassett4 have shown to contain the thyrotropic hormone and with which Loeb and Bassett have produced exophthalmos in guinea pigs. The data are given in the following table.It is clear that aceti...