Miriam E. Simpson

Influence of Hypophysectomy and Growth Hormone on Cartilage Sulfate Metabolism.

Summary Hypophysectomy of immature growing rats results in a markedly diminished sulfate uptake of costal cartilage as judged from the cartilage to plasma inorganic sulfate specific activity ratios. The injection of growth hormone partially restore this ratio to normal under the conditions employed. The plasma inorganic sulfate specific activities of hypophysectomized rats are several fold higher than those of normal rats.

Similarity of response of thymus and lymph nodes to administration of adrenocorticotropic hormone in the rat

Summary Administration of purified ACT to the male rat produces by its action on the adrenal cortex a striking reduction in the weight and size of the thymus gland and the cervical lymph nodes. This same effect is not manifest in the adrenalectomized animal. This finding furnishes further evidence for the existence of a definite relationship between adrenal cortex and thymus and lymph nodes. Functional interpretation of this relationship awaits further experimental and clinical study.

Disturbance of carbohydrate metabolism in normal dogs injected with the hypophyseal growth hormone.

Clinical and experimental data have been accumulating indicating a functional relationship between the anterior lobe of the hypophysis and the pancreatic islets. (See especially Houssay et al.) Due to the importance of such a relationship we think it justifiable to note briefly some remarkable findings bearing on this matter. Two litters of purebred dachshund, one consisting of 2 males, the other of 2 females were secured. One male and one female were injected daily intraperitoneal with the anterior hypophyseal growth hormone, 21 free of gonad stimulating hormone, for a period of about 8 months. Skeletal growth and body weight greatly exceeded that of the litter-mate controls, the weight of the injected animals soon being double that of their controls. Skin overgrowth and folding were present in both but was particularly prominent in the male. After 8 months of daily injection the male developed polydypsia, polyuria, polyphagia and became emaciated. The animal suffered from skin infection with loss of hair. He was inactive and evidently sick. Fehlings test of the urine was strongly positive. Only a trace of albumin was present in the urine. Fasting blood sugar was 232 mg. %. When injection was stopped for one week the animal improved, but rapidly failed again on resumption of injection. At present, 4 months after cessation of injection, the animal still has sugar in the urine. The test for albumin is now negative. The volume of urine has decreased and the animal has almost completely recovered. The female dachshund has now been injected one year. Though responding markedly to the growth hormone she has never shown clinical or laboratory evidence of disturbance of carbohydrate metabolism. Urine and blood sugar have remained normal.

Influence of lactogenic preparations on production of traumatic placentoma in the rat

Summary Normally occurring or artificially induced lutein tissue in the rat does not produce progestin, as shown by the placentoma test. The only pituitary preparation which has been shown to stimulate the production of progestin by such lutein tissue is lactogenic hormone. Therefore, besides its classical mammotrophic activity and crop-sac stimulating function, the “lactogenic hormone’ is important in activation of the corpus luteum, and for this reason should be considered a part of the gonadotrophic complex.

Hypothalamic Control of Pituitary Function and Corpus Luteum Formation in the Rat.

Summary and Conclusions 1. Bilateral hypothalamic lesions ventral to the ventro-medial nucleus in female rats resulted in persistent vaginal cornification in the majority of animals. Thyroid morphology and I131 uptake in obese rats with constant cornification indicated increased thyroid activity. The ovaries of those showing constant cornification contained follicles of all sizes but no corpora lutea. The interstitial tissue was not atrophic. 2. The follicles were readily converted into corpora lutea by administration of luteinizing substances, both human chorionic gonadotrophin and pituitary interstitial cell stimulating hormone. The interpretation is made that secretion of luteinizing factor by the pituitary may be impaired by small, bilateral hypothalamic lesions ventral to the ventro-medial nucleus.

Hormonal Requirements for Pregnancy and Mammary Development in Hypophysectomized Rats.

Summary The same daily doses of purified lactogenic hormone (60 I.U.) and estrone (10 I.U.) capable of inducing beginning lobulo-alveolar mammary growth in the hypophysec-tomized rat, also ensured successful implantation in rats hypophysectomized and injected from the day of sperm. Since only about one-half of the estimated number of implantations were found to have developed normally through mid-pregnancy it follows that these 2 pure substances in the doses used did not adequately substitute for the intact pituitary of a pregnant rat. When injected alone neither lactogenic hormone nor estrone permitted implantation to occur.

Specificity of the Epiphyseal Cartilage Test for the Pituitary Growth Hormone.

Summary 1. Purified anterior hypophyseal hormones other than the growth hormone were administered to hypophysectomized rats for a period of 4 days and measurements made of the width of the uncalcified portion of the proximal epiphyseal cartilage of the tibia. 2. While adrenotropic hormone slightly decreased the cartilage width, thyroxin and the thyrotropic as well as the mammotropic hormones caused slight enlargement of the epiphyseal cartilage. The effect was smaller, however, than the response caused by a single growth hormone unit, at all dose levels tested, and the effect did not show gradation with dosage which is a characteristic of the growth substance.

Relation of Prolan to the Anterior Hypophyseal Hormones.

In our preceding paper we have shown the incapacity of the hormone found in the urine of pregnant women (Prolan) to affect sexual development and function in hypophysectomized animals. It is now well established that Prolan is also for some reason limited in its effect on the gonad development which can be produced within 100 hours in immature normal animals, as measured in terms of ovary weight. We can, for example, state that no matter how frequently or how much Prolan is administered the average weight of the ovaries produced in 26-day-old rats at the close of 4 days of treatment is about 60-70 mg. and the maximum in several hundred of our experiments was 102 mg. We, and other investigators, have pointed out that in contrast with this the implantation of anterior hypophyseal tissue is very markedly more effective in the ovary weights produced in young rat hosts, and we have shown that increasing “doses” of the implants also give increasing weights of the young ovaries. As will be detailed in another communication, similar very marked effects on the immature gonads can now be secured by certain fractions out of extracts of anterior hypophyseal tissue. Here also, in contrast to Prolan, increasing doses give increasing effects on the youthful gonads. With such preparations, we have, for instance, in 100 hours produced ovaries weighing 190 mg. We searched for a point of view which would harmonize these 3 experiences. That Prolan is totally ineffective in hypophysectomized animals and that it is only partially effective in young animals when compared with cases to which actual hypophyseal substances were administered seemed to us best explained by regarding Prolan only as an activator to a substance in the hypophysis itself which we may call a pro-hormone, a substance totally absent, of course, in hypophysectomized animals and small in amount in very young normal animals.

Potent, sterile and low-protein extracts of the growth hormone from the anterior hypophysis

The following procedure has been used in obtaining extracts of the growth hormone of the anterior hypophysis for producing experimental gigantism in rats and provoking the growth of hypophysectomized puppies in the hands of Dr. Frederick L. Reichert. 2,000 gm. of beef anterior hypophysis. The anterior lobes of beef hypophyses had been stored for a period of between 18 and 24 months with solid CO2 in an insulated container in a room held at −5.0° F., after having been ground in a Wiley laboratory mill. 12,000 cc. distilled water. 3,000 cc. 0.2/N Ba (OH)2 Centrifugation: The alkaline mass was mixed by an electric household stirrer (“kitchen mechanic”); then centrifuged in a Sharpies laboratory centrifuge. Large nozzles were used in the first centrifuging in order to remove the bulk of the insoluble material; the extract was then centrifuged at a rate of about 10 cc. per minute. The centrifuge was cooled with salt and ice at all times and the extract was kept surrounded with ice. Neutralization: 0.2/N H2SO4 was added slowly with constant stirring by the same stirrer. Phenol red was used as indicator and the end point chosen was about pH 8.0, about 500 cc. being required. The Ba not already precipitated by H2SO4 was removed by a slight excess of Na2SO4. Centrifugation: Sharples centrifuging was then repeated, finishing as before at a rate of 10 cc. per minute. Filtration: A standard 14 cm. Seitz asbestos filter was used. A pressure of about 10 1hs. per square inch was applied by the use of a tire pump. Time required for filtration of I liters was about 45 minutes; towards the end, the rate of flow through the filter had decreased to half the original rate. Precautions were taken to maintain sterility of the solution after filtration. The extract was stored in 25 cc. lots in rubber-capped bottles and kept frozen until needed for injection.

Pheochromocytomas of adrenals in male rats chronically injected with pituitary growth hormone.

Summary The long term administration of pituitary growth hormone to adult male rats of the Long-Evans strain resulted in progressive growth in body weight and length. Many neoplasms developed in the rats during this prolonged period, but the incidence in treated rats was not significantly different from that in controls, with the exception of tumors of the adrenal medulla. Pheochromo-cytomas occurred in 9 of 16 growth hormone injected rats and in none of the controls.