Ribonucleic acid labelling perfused livers of protein-fed and protein-deprived rats

Several observations suggest an increased RNA synthesis in livers of protein-deprived rats, though the RNA/DNA ratio is decreased. A number of hormones may be involved in these changes. Therefore, we studied in RNA metabolism in isolated perfused livers taken from protein-fed and protein-deprived rats. (3H)-orotic acid was given to the rats 2 h before liver explantation, and (14C)-orotic acid was added to the perfusate. Other rats, called controls in vivo, whose livers were not transplanted were also given (3H)-orotic acid followed by (14C)-orotic acid. The livers of these rats, which were not hormone supplemented, were labelled for the same length of times as the livers in vitro. The ratio specific RNA radioactivity/specific nucleotide radioactivity x RNA/DNA was determined and taken as a measure of the RNA synthesis per liver cell. In the controls in vivo, this ratio was significantly higher for protein-deprived than for protein-fed rats. In livers from the protein-fed rats, labelling in vitro increased significantly when growth hormone, hydrocortisone, insulin and tri-iodothyronine were added to the perfusate. Labelling was also significantly higher in these livers than in the controls in vivo. In livers from protein-deprived rats, the ratio in question was the same whether the hormones were added to the perfusate or not, and was significantly lower than in the controls in vivo. Differences in RNA labelling are thus obtained in our in vitro system. Gel electrophoresis of RNA demonstrated normal RNA labelling, showing that the system is suitable for studying liver RNA synthesis. Further refinement can be made by studying the labelling of UTP and CTP. The results might suggest that the liver from a protein-fed rat, explanted in vitro, may increase its RNA synthesis under the influence of the four hormones in question, and that the RNA synthesis of the liver of a protein-deprived rat is high in-vivo and that it might decrease, when it is explanted to in vitro conditions.     

Radioautographic studies with methionine-3H and cytidine-3H in protein-fed,protein-deprived and starved rats

Summary Male, growing rats were either fed on a protein-rich or a protein-free diet or starved. At various intervals before they were killed, they were given either cytidine-³H or methionine-³H subcutaneously. Radioautographs of several organs were prepared. Grain counts were performed on sections of liver, pancreas, kidney, adrenal, spleen, stomach, duodenum, heart, lung, striated muscle, testis, skin and cerebellum. They revealed inter alia an increased labelling, especially of ribose nucleic acid, in the liver during protein depletion and starvation. These changes were also found in the ribose nucleic acid labelling of nucleoli, nucleolus-associated chromatin and non-nucleolar nucleus in liver cell nuclei. Changes were also found in many other organs. The findings might be interpreted as signs of a shift of nucleic acid and protein synthesis during protein deprivation and starvation from less essential to more essential' organs. — Some aspects of the intracellular ribose nucleic acid and protein metabolism are discussed.The investigation was supported by a grant from the Swedish Medical Research Foundation.     

The ultrastructure of the liver in thyroid-fed rats

Summary Rats were fed on a 25% casein diet or the same diet supplemented with desiccated thyroid. The rats were killed after 16 days. Histological sections of the livers of the control rats show coarse, basophilic inclusions and abundance of glycogen in the cytoplasm. In the thyroid-fed rats there is a diffuse, cytoplasmic basophilia with basophilic rods and no or almost no glycogen. Under the electron microscope large areas of glycogen are to be seen in the cytoplasm of the control animals. Mitochondria and rough-surfaced endoplasmic membranes, often in large stacks, are found together. The liver cells of the thyroid-fed rats have little or no glycogen in their cytoplasm. Mitochondria, endoplasmic reticulum, and free ribosomes and polysomes are evenly distributed all over the cytoplasm. There seems to be an increase in the ratio of free to membrane-bound ribosomes and polysomes in the thyroid-fed rats. The possible significance of this observation in relation to RNA synthesis is discussed.This work was supported by grants from the Swedish Medical Research Council (Project No. K-69-12X-623-05) and the Swedish Cancer Society (Project No. 95-K69-03X).     

Elevated serum growth hormone in protein-deprived rats, and decreased liver RNA after hypophysectomy.

Serum growth hormone was significantly higher in rats fed on a non-protein than on a 25% casein diet. In both dietary groups, hypophysectomy provoked a decrease in the RNA/DNA ratio of the liver, and, especially in the protein-deprived rats, a decrease in the RNA synthesis per liver cell, measured as the product of the RNA/DNA ratio, and the ratio of specific RNA activity to specific nucleotide activity, two hours after administration of [3H]orotic acid. In the protein-deprived rats, full reconstitution of these figures was obtained with a combination of growth hormone, hydrocortisone and triiodothyronine, but not quite with growth hormone alone. The possible role of growth hormone in protein deprivation is discussed.     

CRH and NPY are not decreased in hypophagic, protein-deprived rats

To test if elevated CRH and decreased NPY might account for pituitary-adrenal activity and hypophagia in dietary protein deprivation, rats received normal or protein-free diet, or were food-or weight-restricted to match effects of protein deprivation. Protein or food restriction increased plasma ACTH. However, hypothalamic CRH mRNA was unchanged by protein deficiency and significantly decreased by food restriction when protein intake was > 50% of normal. Arcuate nucleus NPY mRNA increased in rats given protein-free diet, correlating with leptin rather than decreased feeding. We conclude that CRH and NPY gene expression does not explain adrenocortical axis or feeding activity in protein-deprived rats.     

Ring shaped nucleoli in liver cells of rats after treatment with Actinomycin D

Summary The incidence of ring shaped nucleoli was studied by means of light and electron microscopical methods in liver cells of rats injected intravenously with Actinomycin D in a dose of 150 g per kg body weight.Ring shaped nucleoli were most frequent 20 minutes after administration of Actinomycin D. The number of fragmented nucleoli was increased 20 minutes after administration of the drug and did not decrease even after 72 hours.Fibrillar ring like inclusions within nuclei and more perichromatin granules at the periphery of nucleoli were observed after treatment with Actinomycin D.

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Zusammenfassung Die Wirkung von Aktinomycin D (150 g/kg) auf die Leberzellen der Ratte wurde licht- und elektronenenmikroskopisch untersucht. 20 min nach intravenöser Aktinomycin-D-Gabe wurden ringörmige Nukleolen beobachtet. Die Zahl der fragmentierten Nukleolen war 20 min nach der Applikation gesteigert. Ihre relative Zahl änderte sich nach Aktinomycingabe nicht. Nach Behandlung der Tiere mit Aktinomycin D kann man in den Kernen der Leberzellen fibrilläre ringförmige Einschlüsse und eine größere Menge von perichromatin-granules an der Peripherie der Nukleolen beobachten.

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The authors are indebted to M. Mikoláová, V. Povolná, I. erníková and Dr. M. Titlbach for their help, to Dr. J. Chaloupka for providing Actinomycin D and to Professor Busch and Professor Wolf for many helpful suggestions.     

The amount and distribution of water, dry matter, and sugars in the digestive tract of rats fed xanthan gum

Diets containing a nutritionally adequate, high-maltose nutrient mixture and either 4% xanthan gum or 4% cellulose were fed ad libitum to rats. The feeding of this gum increased the combined weight of the small intestine and its contents by 110%. This effect was partially due to an enlarged intestinal cell mass and to extra dry matter in the contents but chiefly to a 400% increase in intraluminal water. Xanthan feeding enhanced greatly the persistence of sugars beyond the proximal quarter of the small intestine and increased their total recovery in the first three quarters of that organ by 150%. The xanthan-induced increase in intraluminal water in the small intestine was partially due to a slowed absorption of osmotically active substances from the gut.     

Satellite nucleoli in the megakaryocytic lineage of rats

The present study was undertaken to provide more information on the incidence of satellite nucleoli in developmental stages of the megakaryocytic lineage. Satellite nucleoli representing solitary silver stained nucleolus organizer regions (AgNORs) present in nuclei in addition to other nucleolar types were observed in all stages of megakaryocytic development. However, the incidence of satellite nucleoli was more frequent in mature megakaryocytes than in less differentiated immature megakaryoblasts and naked megakaryocytic nuclei representing the terminal stages of megakaryocytic development after loss of the cytoplasm transformed to thrombocytes. There is a possibility that the increased incidence of satellite nucleoli in mature megakaryocytes might be due to the loss of AgNORs from active nucleoli characteristic for immature cells. The decreased incidence of satellite nucleoli in naked megakaryocytic nuclei might reflect their disintegration in the terminal stages of the megakaryocytic development.