Changes in rat liver cyclo(His-Pro) binding sites upon castration and testosterone administration

Histidyl-proline diketopiperazine [cyclo(His-Pro)] binding was compared in livers from male and female rats. Cyclo(His-Pro) binding of female rat liver was very much lower than that of male rat liver. Scatchard analysis showed that the sex difference in cyclo(His-Pro) binding was due to different binding capacity. Cyclo(His-Pro) binding of castrated male rat liver was significantly decreased. Testosterone replacement raised the binding to the control level, and an excess of testosterone increased the specific binding beyond the control level. The testosterone-induced changes in cyclo(His-Pro) binding were also due to variation in the binding capacity. These findings indicate that testosterone is an important factor in the regulation of cyclo(His-Pro) binding in the rat liver.     

Effect of castration and testosterone administration on the neuromuscular junction in the levator ani muscle of the rat

Summary The ultrastructure of the neuromuscular junction (n.m.j.) of the androgen-sensitive levator ani muscle was studied in normal adult male rats, in 8-month-old rats castrated at the age of one month and in castrated rats treated with testosterone propionate (TP). Castration does not result in significant changes of the n.m.j. The density of synaptic vesicles and the postsynaptic junctional folds remain practically normal in spite of marked atrophy of the muscle. TP administration for 7 days results in marked changes in preand postsynaptic structures. There is slow progressive depletion of synaptic vesicles, appearance of cisternae and coated vesicles in axon terminals, and coalescence of coated vesicles with the plasma membrane. Coated vesicles are also found inside Schwann cells and among junctional folds. Dense core vesicles appear both in the axon terminals and in the postsynaptic area. Collateral sprouting of terminal axons with the formation of new immature junctions is observed. After 35 days of TP administration depletion of synaptic vesicles continues. Glycogen -particles, mostly freely dispersed, occasionally seen in axon terminals 7 days after TP administration, subsequently increase in number. In the endplate zone of the muscle fibre increased protein synthesis is indicated by a rapid increase in ribosomes and irregularly located myofilaments and myofibrils. The appearance of n.m.j. after testosterone administration resembles that described after nerve stimulation; the degree of change is however less pronounced.The authors wish to acknowledge the skillful technical assistance of Mrs. L. Vedralová     

Proliferation of skeletal muscle satellite cells after castration and administration of testosterone propionate

Thirty-six neonatal pigs were randomly assigned to the following treatment groups: sham implanted gonadally intact males (B), sham-implanted castrated males (C), or castrated males implanted with testosterone propionate (C + TP). Four pigs from each group were sacrificed at 7, 14, or 21 days of age after a 6-hr continuous infusion of [3H]thymidine. Myofibers isolated from the triceps brachii were prepared for satellite cell enumeration by light microscope autoradiography. A developmental decline in labeled myofiber nuclei occurred in all groups, however, the greatest decline occurred in C (P less than 0.01). A treatment-by-age interaction was observed for percentage of labeled nuclei. Castration reduced total and labeled nuclei per millimeter myofiber (P less than 0.05), and C + TP had a higher percentage of labeled nuclei than C (2.8 vs 2.2%; P less than 0.05). Since triceps brachii muscles from 21 day B and C + TP were 120% (P less than 0.05) of C, the results indicate that postnatal growth of skeletal muscle is dependent on satellite cell mitotic activity and that testosterone enhances this activity in neonatal pigs.     

Quantitative analysis of muscle cell changes in compensatory hypertrophy and work-induced hypertrophy.

The cytological characteristics of two modes of muscle hypertrophy were studied in the extensor digitorum longus muscle of the rat. Comprensatory hypertrophy (CH) was produced by tenotomy of the tibialis anterior muscle and work-induced hypertrophy (WIH) was produced by forced swimming of the animal. While both methods produced an increase in muscle weight and cell size, these two parameters did not correlate. Morphometric analyses of the hypertrophied muscle cells demonstrated that in CH-muscle there was an increase in mitochondrial volume density, a decrease in myofibrillar volume density and no change in sarcotubular or nuclear volume density. WIH-muscle demonstrated an increase in sarcotubular volume density but no change in mitochondrial, myofibrillar or nuclear volume density. It is concluded that in CH-muscle, the cell volume increase is attributable to mitochondrial volume increase and that there is no increase in the contratile myofibrillar component of the cell. WIH-muscle, on the other hand, has a cell volume increase which is attributable to a proportional increase in these organelles.     

On the time course of surgically induced compensatory muscle hypertrophication of the rat plantaris muscle.

1. The time course of the hypertrophy and hyperplasia of the rat plantaris muscle was determined from measurements of total muscle mass and cross-section analysis of fixed muscle. 2. Muscle enlargement was induced by the surgical removal of the plantaris synergist muscles, the gastrocnemius and the soleus. 3. From the date of surgery through the third post-operative week, muscle enlargement is due to fiber hypertrophication (approximately 100% increase in diameter). After post-operative week three, muscle enlargement is due to a combination of hyperplasia and hypertrophy. At week four the cross-sectional areas return to control values. 4. The neuromuscular junction area was determined by measuring Karnovsky stained post-synaptic membrane. Only a modest 10-30% increase was noted at weeks 2 and 3 with a return to control levels at week 4. The differences were not statistically different.     

Quantitative studies of compensatory testicular hypertrophy following unilateral castration in the boar

Unilateral castration of Large White X Landrace boars at monthly intervals up to 5 months of age, with the remaining testis being removed 2 months later, resulted in compensatory hypertrophy of the testis which decreased with age. In pigs 3 and 4 months old there was significant hypertrophy of the testis but at 5 and 7 months of age testicular weight of the hemicastrates did not differ significantly from control values. The increase in the testicular weight of unilaterally castrated pigs was correlated with an increase in the number of Sertoli and germ cells at 3 months of age and germ cells at 4 months of age occupying the seminiferous epithelium. This was correlated with increased total seminiferous tubule length and larger cross-sectional area of the tubule. Sertoli cell occupancy did not differ significantly between unilaterally castrated and intact boars.     

Satellite cells of the rat soleus muscle in the process of compensatory hypertrophy combined with denervation

Compensatory hypertrophy was induced in the rat soleus muscle by sectioning the tendon of the ipsilateral gastrocnemius and plantaris muscle. Seven days after tenotomy of synergistic muscles, when soleus hypertrophy attains about 40%, the number of satellite cells (expressed as percentage of all muscle nuclei found in the same cross-sections) as revealed by electron microscopy, was increased from 5.8+/-0.06% in the normal soleus muscle to 16.6+/-1.26%. After four days' denervation of the soleus muscle the percentage of satellite cells was increased to 7.2+/-0.62%. In experiments where hypertrophy of the soleus muscle was combined with denervation three days after tenotomy of synergists, and examined after another four days (during which time it loses, as has previously been shown, over 40% of its predenervation weight), the number of satellite cells was greatly increased to 29.9+/-3.42%. This increase is apparently due to two independent processes which take place during the first postoperative period: a) mitotic division of satellite cells during the early stages of compensatory hypertrophy and b) pinching off of muscle nuclei from rapidly atrophying muscle fibres due to subsequent denervation. Activation of satellite cells was mainly manifested by expansion of smooth and especially of rough endoplasmic reticulum, a rich Golgi complex, high pinocytotic activity, increased number of ribosomes and by nuclear changes. Concomitantly with the increased number of satellite cells, proliferation of fibroblasts, macrophages and mast cells could be observed.     

Time course adaptations in rat skeletal muscle isomyosins during compensatory growth and regression

The purpose of this study was to ascertain the time course of change during both compensatory growth (hypertrophy) and subsequent growth regression on myosin isoform expression in rodent fast-twitch plantaris muscle in response to functional overload (induced by removal of synergists). Peak hypertrophy of the plantaris muscle (92%) occurred after 9 wk of overload. After 7 wk of overload regression (induced by a model of hindlimb unweighting), muscle weight returned to within 30% of control values. Myofibril protein content (mg/g muscle) remained relatively constant throughout the overload period but became significantly depressed relative to control values after 7 wk of regression. However, when expressed on a per muscle basis (mg/muscle) no differences existed at this time point (t = 7 wk regression). The distribution of native myosin isoforms in the myofibril protein pool of the overloaded plantaris muscle reflected a progressive increase (23% at t = 9 wk; P less than 0.001) in the relative proportion of slow myosin (Sm). This change was also accompanied by increases in intermediate myosin (Im) as well as the repression of the fast myosin one (Fm1) isoform (P less than 0.001). These shifts in Sm and Fm1 isoform expression were gradually reversed during the regression period, whereas Im remained elevated relative to control values. These adaptive changes in myosin isoform expression during both hypertrophy and regression were further supported by concomitant shifts in both myosin adenosinetriphosphatase (ATPase) activity (decreased during overload) and slow myosin light chain (SLC) expression. However, during regression the changes in myosin isoform expression and myosin ATPase were not as synchronous as they were during overload. Estimation of the mixed myosin heavy chain (MHC) half-life (t 1/2), using a linear model that assumes zero-order synthesis and first-order degradation kinetics, revealed t 1/2 values of approximately 19 and 10 days for the overload and regression periods, respectively. Collectively these data suggest that 1) skeletal muscle myosin isoforms and corresponding ATPase activity are in a dynamic state of change, although not completely synchronous, in response to altered muscle stress, and 2) the kinetics of change in the mixed MHC protein pool are slower during compensatory growth compared with regression of growth.     

Distribution of 14 elements in various rat tissues following hypophysectomy,thyroparathyroidectomy, adrenalectomy,and castration

The tissue distribution of 14 elements was simulatneously determined in rats 28 d after hypophysectomy (HPY), thyroparathyroidectomy (TPTY), adrenalectomy (ADY), and castration (CTN). The elements Na, K, Ca, Mg, Fe, S, P, Rb, Sr, Mn, Cu, and Zn were investigated in whole blood, plasma, brain, liver, kidney, heart, skeletal muscle, and bone. Additionally Mo was determined in kidney and liver. The following results were obtained: 1) With regard to hormone deficiency: HPY induced the most noticeable, variations on all the elements tested owing probably to the direct and indirect effects of adenohypophyseal hormones. ADY led to the expected modification of Na and K but also to a Sr accumulation and a Rb depletion. TPTY induced a sharp decrease in plasma and tissues Ca, an increase in plasma P, but did not disturb the two elements in bone. An increase of Rb in many tissues and of Fe in heart, kidney, and liver were also observed. CTN had little consequences except in bone whose Cu and Fe contents were increased: 2) With regard to element variations: K, Mg, and S underwent little change. Discriminations were revealed between elements such as K and Rb, Ca and Sr, Ca and Mg, and Cu and Zn. The changes of Rb and Sr were consistent with regulatory mechanisms. The accumulation of Fe and Cu in tissues such as liver after HPY, TPTY, and ADY, suggest that the hormonal deficiencies could worsen the hemochromatosis with Wilson's disease; 3), With regard to plasma and tissues: No correlation appeared in element levels between plasma and other tissues. Brain was the least affected and liver, kidney and bone the most.     

Purification, molecular properties and biosynthesis of a specific protein component induced under compensatory hypertrophy in the rat skeletal muscle

The compensatory hypertrophy in the rat skeletal muscle was induced by the method employing tenotomy and the protein components of the hypertrophied muscle were compared with those of the control muscle by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A protein component, referred to as 64-kDa protein, increased in quantity in the hypertrophied muscle. 64-kDa protein was purified to homogeneity from the extract of the rat skeletal muscle and its molecular properties were studied. Based on criteria which include molecular weight, amino acid composition and immunochemical reactivity, 64-kDa protein was indistinguishable from rat serum albumin. In vitro culture of muscle cells provided evidence showing that 64-kDa protein is synthesized in the muscle cells and rate of its synthesis is significantly greater in the hypertrophied muscle.     

Steroid glucuronyltransferases of rat liver. Properties of oestrone and testosterone glucuronyltransferases and the effect of ovariectomy, castration and administration of steroids on the enzymes.

1. Microsomal preparations from rat liver, kidney and intestine were tested for UDP-glucuronyltransferase activity by using oestrone, oestradiol-17 beta, oestriol, testosterone, cortisol, cortisone, corticosterone, aldosterone, tetrahydrocortisol and tetrahydrocortisone as substrates. The microsomal preparation from the liver glucuronidated oestrone, oestradiol-17 beta and testosterone. 2. The specific activity of the enzyme was significantly higher in livers from female rats than in those from male rats. 3. Testosterone was actively glucuronidated by both sexes. Cortisol, cortisone, corticosterone, aldosterone, tetrahydrocortisol and tetrahydrocortisone were not glucuronidated by any of the three tissues. 4. The non-ionic detergent Lubrol WX activates liver microsomal UDP-glucuronyltransferase 2-3-fold with oestrone and testosterone as substrates. 5. Oestrone glucuronyltransferase was inhibited by oestradiol-17 beta, predominantly competitively and by testosterone non-competitively. Bilirubin was a non-competitive inhibitor of oestrone glucuronidation. p-Nitrophenol had no effect. 6. Oestrone glucuronyltransferase could not be stimulated by either acute or prolonged treatment of animals with phenobarbital, whereas a single dose of 3-methylcholanthrene led to a moderate stimulation. 7. Ovariectomy leads to a 56% decrease in oestrone glucuronyltransferase activity; administration of oestradiol-17 beta induces the enzyme to normal activity after 12 days, and after 15 days the activity is twice the control value. Actinomycin D and cycloheximide block the oestradiol-17 beta-induced increase in enzyme activity. 8. Castration has no effect on the activity of testosterone glucuronyltransferase, nor does administration of testosterone influence enzyme activity. The results provide strong evidence for the existence of multiple steroid glucuronyltransferases in the liver of the rat.     

Impairment of renal compensatory hypertrophy by hypothyroidism in the rat

Renal compensatory hypertrophy (RCH) occurs in hypothyroid rate, but it is impaired when compared to RCH found in euthyroid controls. It is due to cellular hypertrophy as the DNA content does not change and the Protein/DNA ratio increases in the compensating kidney. RCH is enhanced by thyroxine (T4) with a rise in the DNA content of the compensating kidney, but the Protein/DNA ratio does not change indicating that hypertrophy is as important as hyperplasia. Corticotrophin (ACTH) given to eu and hypothyroid rats enhances RCH with an increase in the protein content of the compensating kidney without any change in its DNA content. In the hyperthyroid rats, the enhanced RCH is not further increased by ACTH and the rise in the kidney DNA content elicited by T4 is suppressed by ACTH. The Protein/DNA ratio is increased by ACTH in hypo, eu and hyperthyroid rats. The renotrophic action of ACTH is due to hyperadrenocorticism: it is related to an increased plasma testosterone level and to a disturbed Na+, K+ and glucose metabolism.