Die DNS-Synthese in Leber und Niere junger Ratten. Autoradiographische Untersuchungen mit3H-Thymidin

Summary Autoradiographic studies following a single i.p. injection of³H-thymidine were performed in liver and kidney parenchyma of new-born and young adult rats at different ages (1, 2, 4, 7, 12, 18, 24, 30, 60 and 120 days). In 1-day-old animals the tritium index (i.e. percentage of DNA synthesizing nuclei) of both organs is lower than in the rat embryo. From 1–4 days fluctuations occur, then the tritium index rises at 7 and 12 days. But now an exponential decrease is observed up to day 120. This depression and fluctuation of the tritium index probably depends on a postpartal functional transposition and especially in the liver on a structural transformation. The mean grain density in the labelled nuclei increases from 1 up to 24 days, which is likely connected with a very high amount of endogenous thymidine in new-born rats. From day 24 the mean grain density can be taken as a relative measure for the rate of DNA synthesis. The reduction in the rate of DNA synthesis from 24–120 days is explained as a consequence of the commutation from the rapid to the slow mode in cellular proliferation.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

DNA synthesis in the nuclei of differentiating muscle fibers of the silkworm,Bombyx mori L.

Summary Incorporation of³H-thymidine (³HTdr) into the nuclei of myofibril-containing myofibers of larvae of the silkworm,Bombyx mori, was shown by means of light microscope (LM) and electron-microscope (EM) autoradiography. The number of DNA-synthesizing myonuclei attains 42% 12–18 h after each molt. Thus in the developing silkworm DNA replication and myofibrillogenesis are coexisting and not mutually exclusive processes as is the rule in vertebrate somatic myogenesis.     

Autoradiographische Studien über die DNS-Synthese im Hypophysenvorderlappen der Ratte

Summary By autoradiographic studies with H³-thymidine it is demonstrated that the percentage of labelled nuclei is very low in the anterior pituitary of normal untreated rats and of rats 14 days after thyreoidectomy. Following castration, however, a marked increase of DNA-synthesizing cells occurs as well as an increase of mitotic figures.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft und das Bundesministerium für Wissenschaftliche Forschung.     

Association of DNA replication with nuclear membrane in larvae ofChironomus thummi

Summary In the larvae ofChironomus thummi, the newly replicating DNA has been found to be associated with the nuclear membrane, as evidenced by the isolation of DNA nuclear membrane complexes (M-band) of³H-thymidine labelled larvae.Acknowledgment. The authors thank Dr D. S. Pradhan for discussions and encouragement.     

Nucleolus DNA synthesis inVicia faba root-tip meristems

Summary Autoradiographs ofVicia faba lateral root-tip meristems were prepared following a 30 min pulse with³H-thymidine. 1/3 of all interphase nuclei in the meristem are labeled, most with a uniform distribution of silver grains. 2–5% of labeled nuclei show specific nucleolus DNA labeling.     

Zum Proliferations-Stoffwechsel der Rattenleber nach Teilhepatektomie. Autoradiographische Untersuchungen mit3H-Cytidin,-Phenylalanin und -Thymidin

Summary During the first wave of parenchymal liver regeneration in adult rats after partial hepatectomy, the cellular synthesis and migration of RNA and the metabolism of protein were studied by autoradiography following an injection of³H-cytidine or³H-l-phenylalanine and double injections of 1 of these precursors +³H-thymidine. The following results were obtained: the synthesis and migration of RNA and the metabolism of protein are enhanced under these conditions of proliferation. In spite of this, the relation of metabolic activity in nucleolus, karyoplasm and cytoplasm remains constant. By double injection techniques it is proved that no differences exist in migration of RNA into the cytoplasm and cytoplasmic protein synthesis between cells with or without DNA synthesizing nuclei.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Continuous labelling method for autoradiographic analysis of cell cycle parameters in steady state cell systems

Summary Under certain conditions, continuous availability of³H-thymidine in steady-state cell systems results in a linear increase of the fraction of labelled cells, the equation of which can be used to determine cell cycle parameters of the system investigated.This research was supported by the Deutsche Forschungsgemeinschaft.     

Relationship between DNA and cholesterol synthesis in kidney after refeeding

Summary DNA and cholesterol synthesis were investigated in the kidneys of fasted-refed rats. Refeeding resulted in an increase in kidney DNA synthesis, as measured by³H-thymidine incorporation, starting at 72 h. The increase in DNA synthesis was accompanied by a stimulation of cholesterol synthesis, as measured by¹⁴C-acetate incorporation into cholesterol.     

Autoradiographische Bestimmung der mittleren Verweilzeit neutrophiler Granulozyten im Blut der Ratte mittels Dauerinfusion von3H- Thymidin

Summary Labelling of neutrophil granulocytes in the blood and sputum of rats was followed by autoradiography for 120 h after starting a continuous infusion of³H-thymidine).     

Cell cycle changes after denervation of the newt limb regenerate,studied in vitro

Summary The proliferation of the mesenchyme of medium bud stage blastemas ofPleurodeles, measured by³H-thymidine incorporation and mitotic index, decreases about 40–50% under denervated conditions when cultivated in vitro for 4 days; epidermis is not affected in this case. Autoradiography of blastemas after³H-thymidine long term labeling shows that 3/4 of the mesenchymal cells and 1/4 of the epidermal cells are cycling when the blastema is innervated; there is no significant change of these percentages when the blastema is denervated. The results show, contrary to in vivo experiments, that denervation does not provoke an exiting from the cell-cycle but only lengthening of the cycle of the mesenchymal cells (probably of the G₁ phase).     

Mechanism of age-dependent involution in embryonic chick notochords

To study the possible mechanism of the age-dependent involution of the notochord, isolated mesenchymefree notochords of chick embryos were cultured in vitro and compared with their counterparts in vivo. Two different aspects were evaluated: (1) DNA synthesis measured by [³H]thymidine incorporation and visualized by autoradiography and (2) cell death quantified by counting the number of pyknotic nuclei. The results demonstrate that [³H]thymidine uptake by notochords shows an age-dependent decrease in vitro as well as in vivo. The number of [³H]thymidine-labelled notochord cells, however, is higher in vitro than in vivo. At the same time, there is an age-dependent increase in pyknosis in the notochord in vivo and in vitro. So, during the aging process, the number of both pyknotic nuclei and of [³H]thymidine-labelled nuclei suggest a high turnover of notochord cells in vitro. From these results, we can conclude that the process of involution in aging notochord seems to be controlled by a programmed intrinsic process, which might be influenced partially by the microenvironment in vivo.     

Einbau von H3-thymidin in zellen und gewebe vonHelix pomatia L. (gastropoda,pulmonata)

Summary It has been demonstrated by injections of H³-Thymidine into adult and juvenile snails (Helix pomatia) that the mitotic activity in the central nervous system, the connective tissue and epidermis was generally higher in juvenile animals than in adult ones. Data are given for the amount of triated thymidine labelled nuclei in the tissues, which are investigated.     

Decrease in [3H]ouabain binding sites in heart and brain from spontaneously hypertensive rats

Summary A decrease in the number of binding sites (B_max) for [³H]ouabain was observed in the heart (37%) and the brain (22%) of spontaneously hypertensive rats (SHR) when compared with age-matched control Wistar Kyoto rats. No variation was detected in the affinity constant (K_D).     

Endogenous oscillatory activity and TTX-sensitive spikes of the heart muscle in early juveniles of the isopod crustaceanLigia exotica

The heart beat of early juveniles of the littoral isopodLigia exotica occurred at a frequency of 250 to 350/min, associated with rhythmic activity of the heart muscle. Each burst was composed of a slow depolarizing potential with superimposed spike potentials. The spike potential was eliminated by perfusion with TTX-containing or Na⁺-free saline. In TTX-saline, the slow potential was unchanged in frequency and amplitude. By current injection into the heart muscle, the rhythm of the slow potential was phase-shifted and its frequency was changed in a membrane potential-dependent manner. These results show that the heart ofLigia early juveniles acts as an endogenous muscle oscillator generating oscillatory slow potentials and Na⁺-dependent spikes.     

Small lymphocyte production and lymphoid cell proliferation in mouse bone marrow

Summary In mice given³H-thymidine systemically during temporary circulatory occlusion of one hind limb, comparison of the labeling of rapidly-renewing small lymphocytes in the tibial marrows demonstrated that these cells were locally produced. Labeling by³H-thymidine infusion revealed that many marrow large lymphoid cells, presumptive small lymphocyte progenitors, had a marked proliferative activity and rapid turnover which varied according to cell size, was maximal in young mice and declined with increasing age.This work was supported by a grant from the Medical Research Council of Canada. The technical assistance of Mrs E. D. Watson is acknowledged.     

Effect of epidermal growth factor on growth and maturation of fetal and neonatal rat small intestine in organ culture

Summary Small intestinal explants from pre- and post-natal rats were incubated in an organ culture system in the absence and presence of epidermal growth factor (EGF). The rate of synthesis of small intestinal DNA and protein as well as the activity of lactase and alkaline phosphatase increased rapidly between 17 and 20-day gestational age, whereafter they declined. The maximal incorporation of³H-thymidine and¹⁴C-alanine into DNA and protein, respectively, was significantly stimulated by EGF (100 ng/ml). EGF had no effect on the activity of either lactase or alkaline phosphatase in the small intestinal explants.Acknowledgment. The project was supported by grants from the Veterans Administration Research Service. The authors wish to thank Dr. M. C. Geokas, Chief, Department of Medicine, Veterans Administration Medical Center, Martinez, CA, for providing us with excellent laboratory facilities and for his encouragement in this study.     

Zur kompensatorischen Nierenregeneration. Autoradiographische Untersuchungen nach3H-Thymidin-Dauerinfusion

Summary By continuous infusion of³H-thymidine, autoradiographic studies were performed to compare cellular proliferation in the compensatory regenerating kidney of rats after contralateral nephrectomy or contralateral temporal (1 h) ischaemia with physiological cell renewal in controls. Under each experimental condition, the highest labelling index is observed within the stratum labyrinthicum and then a gradual decrease occurs from there to the stratum subcorticale, zona intermedia and basalis. As to the topic of DNA synthesizing cells in the different zones of the kidney, there are only quantitative differences between physiological cell renewal of the controls and compensatory regeneration following contralateral nephrectomy and contralateral ischaemia.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

A differential effect of L-serine on the incorporation of3H-deoxycytidine and3H-thymidine into DNA of rat thymocytes

Summary The addition of L-serine to short-term cultures of rat thymocytes stimulated the incorporation of³H-deoxycytidine into DNA, but simultaneously depressed the incorporation of³H-thymidine into DNA.This investigation was supported by NIH grants No. CA-10291 and T12CA8021 from the National Cancer Institute.     

Untersuchungen über den Wirkungsmechanismus von Procarbazin (Natulan®)

Summary The cytotoxic methylhydrazine derivative procarbazine or Natulan® has a marked influence on the synthesis of deoxyribonucleic acid in Ehrlich ascites carcinoma cells. The incorporation of³H-thymidine into DNA — measured by autoradiographic methods — is inhibited.     

Thiamine transport by human intestine in vitro

Summary Surgical specimens of human gastrointestinal mucosa and muscle were incubated in vitro with thiaminethiazole-2-¹⁴C. Labelled thiamine uptake was uphill in mucosal tissues and downhill in muscle. Small intestinal mucosa accumulated labelled thiamine in a phosphorylated form, while gastric and colonic mucosa, as well as muscular layers of all gastrointestinal segments studied, did not.This work was supported in part by a grant from the Italian Research Council, Rome, Italy.