The pituitary-testicular axis was investigated in the streptozotocin diabetic male rat to determine the relationship between hormonal alterations and Leydig cell morphology. Male Wistar rats (60 days of age) were made diabetic with a single intravenous injection (65 mg/kg body weight) of streptozotocin and were studied with non-diabetic controls at 7–11, 14, 21, and 28 days after treatment. The observations on these animals were compared to those from diabetic rats (28 days) treated with 1–2 U protamine zinc insulin/100 mg body weight. Rats with untreated diabetes had hyperglycaemia and weighed less than controls. Untreated rats also had low plasma testosterone levels associated with decreased prostate and seminal vesicle weights. Insulin treatment partially or completely prevented these abnormalities. Studies of testosterone production y perfused testis suggested Leydig cell insensitivity to LH as one possible cause of decreased androgen blood levels in diabetic rats. Lipid droplets not found in Leydig cells of normal rats were seen in diabetic animals. Plasma FSH levels were also low in untreated diabetic rats but were normal in insulin

The pituitary‐testicular axis in the streptozotocin diabetic male rat: evidence for gonadotroph, Sertoli cell and Leydig cell dysfunction

Ventral prostates taken from orchiectomized mice show no detectable development when cultured for four days on a chemically defined basal medium, but show extensive development if insulin is added to the basal medium.1 In these earlier studies with retrogressed prostates,1' 2 insulin added in vitro enhanced the incorporation of C14-amino acid into protein, increased the incorporation of labeled precursor into ribonucleic acid, and promoted histological repair of cells in the secretory epithelium. In contrast, under these same conditions of culture, testosterone, a potent hormonal stimulant for the prostate in vivo, gave no demonstrable repair of the retrogressed organ in vitro. The present experiments with prostates from orchiectomized donors demonstrate for the first time that testosterone added in vitro can evoke two of its principal physiological effects, namely, augmentation of citrate secretion and stimulation of protein synthesis, provided that insulin also is added to the culture medium. M\\Ioreover, testosterone added alone in vitro augments both protein synthesis and citrate secretion in partially repaired prostates. M3faterials and Mlethods.-iiale mice of the Swiss white strain weighing 25-27 gm were orchiectomized at 9 weeks of age; the testis, epididymis, and surrounding fat pad were removed, together with as much of the vas deferens and associated fat as feasible. Three weeks after gonadectomy,' mice were sacrificed by decapitation. Ventral prostates were dissected free of the urethra and placed in culture for 4 days at 360; the gas phase was 5% C02-95% 02. After the first 2 days in culture, explants were transferred to fresh medium and cultured for an additional 48 hr; in experiments designed to assess the effect of the hormones on protein synthesis, this second medium contained DL-leucine-1C14 (0.055 ,c/ml). Cultures were randomized, and each entry in Tables 1-3 is a comI)osite of pooled data from two or three experiments; explants were so distributed that a proportionate number in each group was tested against a similar number in every other group. Culture medium: The basal medium was made up of 4 parts Waymouth's tissue culture medium,4 prepared from stable stock solutions,' and 1 part agar (a freshly autoclaved preparation containing 165 mg agar/10 ml of glass-distilled water). Concentrated solutions of the hormones6 and of leucine-1-C\"4 were prepared as follows: DLleucine-1-C14 (1 mg/ml in 0.03 N HCl); insulin (1 mg crystalline zinc insulin/ml in 0.03 N HCI); testosterone (an aqueous suspension, 0.31 mg/ml of buffer solution to which Tween 80 had been added).7 Determination of citrate; prostates partially repaired: Preliminary studies disclosed that the amount of citrate formed by retrogressed prostates during 4 days of cultureeven though two or three glands were pooled-was too small to be measured; hence, only partially repaired organs were used in these studies. Prostates were taken from orchiectomized mice that had been pretreated with androgen; two subcutaneous injections of testosterone lpropionate (500 Ag/O.10 ml in aqueous suspension) were given, one injection at 96-100 hr and the other at 48-52 hr prior to sacrifice. Two prostates were used for each citrate determination. Explants were taken off the medium, placed in homogenizing tubes and quickly frozen by plunging the tubes into a cold ethanol bath to which (Irv

Regulation by testosterone and insulin of citrate secretion and protein synthesis in explanted mouse prostates.

Male rats rendered diabetic by IV streptozotocin (65mg/kg body weight) were treated with exogenous insulin or testosterone Charcoal-coated dextran and polyacrylemide gel electrophoresis techniques were employed in studying the characteristics of androgen (R1881) binding to prostate cytosol protein In comparison with normal (N) rats, the replacement therapy of diabetic (D) animals with insulin (D+I) or testosterone (D+T) was able to restore epididymal weight (N = 040 ±004 g; D = 018 ± 002 g; D+I = 042 ± 005 g; D+T = 040 ± 006 g) and total prostate weight (N = 024 ± 002 g; D = 015 ± 002 g; D+I = 024 ± 005 g; D+T = 035 ± 006 g) Testicular endogenous content of testosterone was restored after insulin treatment (N = 154 ± 13 ng/testis; D = 41 ± 5 ng/testis; D+I = 142 ± 9 ng/testis), and significant improvements of serum testosterone levels were also achieved (N = 540 ± 64 ng/100 ml; D = 238 ± 37 ng/100 ml; D+I = 358 ± 18 ng/100 ml) Prostate cytosol of streptozotocin-diabetic rats had strongly lowered capacity for 3H-R1881 binding compared with controls (94 and 12 fmol/mg protein, respectively) Testosterone treatment produced a 33-fold improvement of this lowered value, whereas the increment seen with insulin was less (15-fold) It is emphasized that some of the improvements caused by insulin replacement therapy in diabetic animals are due to the partial restoration of testosterone secretion Thus, the combined actions of insulin and testosterone (instead of insulin alone) seem to be of major importance in the maintenance and regulation of accessory sex glands function

/pdf/androgen-receptors-in-the-diabetic-rat-3wcfbi5yq3.pdf

Androgen receptors in the diabetic rat.

Chapter 14 – The Prostate Gland in Organ Culture

Publisher Summary This chapter discusses the morphological and biochemical parameters of androgen effects on rat ventral prostate in organ culture. The rat ventral prostate is a typical target organ for androgens. The ventral prostate gland of the adult rat consists of ducts and alveoli separated by thin strands of fibromuscular stroma and lined with cuboidal or mostly columnar secretory epithelium, which often projects in folds into the lumen of alveoli. The columnar cells exhibit a well-defined supranuclear clear zone corresponding essentially to the Golgi apparatus. A time–course study investigates the action of testosterone added to insulin-containing medium. It shows that, in the surviving explants kept in the absence of androgen, protein synthesis decreases for 2 days and then increases. The effects of insulin and testosterone initially observed in maintenance experiments with total protein and RNA are confirmed in various ways. The results of electrophoretic analyses indicate that the increased incorporation of radioactivity into protein is certainly not only a matter of amino acid pool, which could not produce such uneven changes of the different proteins as those observed.

Morphological and Biochemical Parameters of Androgen Effects on Rat Ventral Prostate in Organ Culture

Effect of insulin and lack of effect of testosterone on the protein of ventral prostates from castrated mice maintained as organ cultures.

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Effect of insulin and lack of effect of testosterone on the protein of ventral prostates from castrated mice maintained as organ cultures.