Effect of the alkylating agent dipin on the induced proliferation of hepatocytes. I. The kinetics of the cell population]

The alkylating drug dipin was injected to mice 2 hours before a partial hepatectomy. Liver regeneration was characterized by a decrease of the intensity of 3H-thymidine label, an increase of the labeled cell index, absence of mitoses, constant number of binuclear cells. The analysis of these data has shown that dipin causes a sharp (more than by 2 times) increase of the S-period and prolonged (up to 6--20 days) blocking of cells in the G2-period. No phenomenon of unbalanced growth was recorded. No changes in duration of prereplicative period, or in the volume of proliferative pool were recorded. The increase of mitotic cycle periods resulted in the cell population synchronization: by the end of the second ay more than a half of hepatocytes were in S-period, by the end of the third day about 80% of cells passed to G2-period.     

Centrioles in the cell cycle of L cells

The structure of centrosome in non-synchronous L-cells culture during the cell cycle has been studied. In mitosis, mother and daughter centrioles, which differ in their ultrastructure, are located perpendicularly in the pole of the spindle. Microtubules, meeting in the pole area terminate mainly in electron-dense clottings of fibrillar matter surrounding the diplosoma. In telophase, disjunction of mother and daughter centrioles begins. At the beginning of G1-period, centrioles move off from each other for several micron, and then draw together again without forming diplosome. Pericentriolar satellites form on mother centriole of some cells at this time, they disappear at the beginning of S-period, replication of centrioles begins; daughter centrioles reach the size of mother centrioles in anaphase. During growth and maturation, centrioles in L-cells undergo structural changes similar to those described for SPEV cells (Vorob'ev, Chentsov, 1982). Several types of meeting points for microtubules exist in L-cells during the whole interphase: surface of centrioles per se, pericentriolar satellites, free foci.     

Structure of the centriolar complex in the hepatocytes of intact and regenerating mouse live

The structure of the cellular center in polyploid hepatocytes of intact and regenerating liver of adult mice has been studied. It was shown that the structure of the centriolar complex depends on stages of the cellular cycle. No pericentriolar structures (such as satellites, appendages and others) and cytoplasmic microtubules were found in the centriolar complex within G0-period. The satellites and appendages are formed in the half of the centrioles within G1-period. The microtubules can branch off some satellites; the daughter centrioles begin to form within S-period; there are diplosomes in the cells within G2-period, some mother centrioles are surrounded with the fine fibrillar halo. It is concluded that the structure of the centriolar complex within G0-period is distinguished by that within G1-period. The structure of the centriolar complex in polyploid hepatocytes has the same feature of reorganization in certain interphase periods of the cell cycle as in diploid cells of some cultured cells and the thyroid epithelium.     

Stimulation by insulin of DNA synthesis in cultured mouse fibroblasts]

With the aid of autoradiography, the effect of insulin on entering S- from G1-period of the mitotic cycle and on the rate of DNA synthesis of the mouse fibroblasts (L), was studied,--in the cells incubated for 24 hr in serum-free medium. In these conditions the cells were temporarily blocked in G1-period. Insulin (100 mcU/ml) increased by 1.5-fold the amount of cells in S-period as well as caused a marked stimulation of DNA synthesis.     

Der Zellzyklus in Fibroblastenkulturen vom Menschen

Summary Autoradiography using H³-Thymidin and Feulgen photometry of nuclear DNA-content were carried out on human euploid fibroblast cultures.The average durations of S- and G2-periods are about 9 hours and 5 hours respectively, with only minor variation. The duration of G1-period may vary from a few hours to more than 20 hours.A combined study of H³-thymidin uptake and Feulgen photometry on the same cell nuclei showed that all cells whose DNA-content places them into the S-period are labelled when they are fixed immediate after a H³-thymidin puls. After continuous H³-thymidin uptake for 5 hours, all S- and G2-nuclei are labelled, whereas the G1-nuclei remain unlabelled.The Feulgen histogramm as well as grain counts over labelled nuclei indicate a constant rate of DNA synthesis during S-period.     

Responses of rat left ventricle cardiomyocytes to constant 2G-hypergravity

2G-hypergravity during 33-days led to hypertrophy of cardiomyocytes (CMC) in rat left ventricle accompanied by capillary number and diameter growth. CMC ultrastructure was normal at all experimental stages. Number of intermitochondrial junctions (IMJ) rose after 33 days of 2G in subsarcolemmal pericapillary zone of CMC, indicating probably to enlarged CMC energy needs. Changes in myocardium achieved during 33 days of 2G were reversible at 39 days of 1G (except IMJ number). Repeated 5-days 2G influence (after 33 days of primary 2G influence followed by 39 days of 1G) led again to CMC hypertrophy and to IMJ number rise in all zones of mitochondria localisation. Primary 5-days 2G influence didn't result in CMC hypertrophy, IMJ number rose only in subsarcolemmal pericapillary zone.     

Structure of the centriole complex in mouse hepatocytes in the experimental induction of a G2 block

In the regenerating liver hepatocytes the centriolar cycle is retarded corresponding to the delay of the nuclear cycle up to the beginning of G2-block. A prolonged staying of cells in the premitotic condition results in the phenomenon that according to their DNA these cells correspond to the G2-period, whereas according to their centriolar complex structure they move into the following G1 and G0-periods, passing mitosis. Thus, in the G2-blocked hepatocytes there is a separation of the nuclear cycle and centriolar cycles. Moreover, during the diping action the centriolar capacity of forming cytoplasmic and mitotic microtubules is suppressed.     

Differentiation of mucous neck cells into parietal cells: a new concept of mitochondrial biogenesis

Parietal cells of the gastric fundic mucosa are small and contain only a few tiny mitochondria when they begin to differentiate from mucous neck cells. The canalicular ATPase activity characteristic of mature parietal cells is discrete in these young cells, whereas areas of very high activity are apparent in the Golgi complex, reticulum, nuclear envelope, mitochondrial wall, and plasma membrane. Close relations and contacts occur between mitochondria and these organelles, and the size and number of mitochondria increase progressively. These relations, as well as mitochondrial ATPase activity (a true differentiation marker), cease once the mitochondria become as numerous and large as those of a mature parietal cell. Our observations suggest that a secondary form of mitochondrial biogenesis, involving the massive participation of other organelles and independent of the classical mechanisms inherent in mitosis, occurs in parietal cells at the beginning of G1 phase during the 6 days of their maturation.     

Dynamics of 3H-thymidine and 3H-deoxycytidine incorporation into the nuclei of a rabbit kidney cell culture during the S period

Using a combined cytophotometric-autoradiographic method a study was made of 3H-thymidine and 3H-deoxycytidine incorporation rates into the interphase nuclei of rabbit kidney cell culture during the S-period. The rate of 3H-deoxycytidine (10(-4) M--10(-6) M) incorporation into nuclei increases throughout the first part of the S-period and decreases from its middle to the end. The patterns of variations of 3H-thymidine and 3H-deoxycytidine incorporation rates into the nuclei of cultured rabbit kidney cells during the S-period were identical.     

Morphometric study of nucleolus-organizing regions of chromosomes from swine embryo kidney cells during Go-, G2-period and mitosis

Quantitative characteristics of nucleolus-organizing regions of chromosomes (NORs, or fibrillar centers, FCs) and some other nucleolar components have been studied with the aid of complete series of ultrathin sections of PK-cells. It has been found that: 1) the number of FCs per cell in the G0-period, in the G2-period and at metaphase is equal to 7.0, 33.7 and 8.0, respectively; 2) volumes of individual FCs in the G0-period (0.033 micron 3), G2-period (0.014 micron 3) and at metaphase (0.025 micron 3) are different; 3) the total volume of FCs, calculated for a haploid set of chromosomes, do not differ in the G0 (0.105 micron 3) and G2 (0.107 micron 3) periods, but exceed twice the FCs volume at metaphase (0.04-0.05 micron 3). These data show that the activation and inactivation of ribosomal genes in interphase PK-cells are not accompanied by a change in the total volumes of FCs and are probably connected with the "fragmentation" and fusion of FCs. Complete inactivation of ribosomal genes at mitosis leads to a decrease in the total volumes of FCs; 4) the nucleolus volume is proportional to the volume of the dense fibrillar RNP-component; in the G2-period the nucleolus volume also correlates with the number of FCs (r = 0.99); 5) the volume of the dense fibrillar component within individual fibrillar complexes--the structures corresponding to one nucleolus-organizing region--is not constant. This is an indirect evidence for the differences in the functional activity of NORs of different chromosomes.     

麦蛾柔茧蜂幼虫唾液腺的显微与超微形态观察

本文采用解剖学观察、显微摄影、透射电镜等方法对麦蛾柔茧蜂Habrobracon hebetor幼虫唾液腺的显微形态、超微结构以及发育特性进行了观察和分析。麦蛾柔茧蜂幼虫唾液腺为一对无色透明至乳白色的管状腺体,自口腔沿中肠两侧向后延伸,单侧腺体在中部先分支、后合并成一不规则环状,端部呈单盲管状。唾液腺管道长度随幼虫龄期增加而呈线性增长。对唾液腺切片进行超微结构观察,发现腺管由两类差异明显的单层细胞组成,I型细胞微绒毛层较厚,胞内除有丰富的内质网和线粒体之外,还含有大量囊泡,并观察到囊泡运输分泌颗粒的现象;II型细胞微绒毛短,胞内的内质网和线粒体数量丰富。本文研究为深入探究寄生蜂幼虫的消化生理以及寄生蜂-寄主互作机制奠定了基础。     

Study of the chondriome in SPEV cell culture after treatment with actinomycin D

It is shown that the 12-hour treatment of cell with actinomycin D (AMD) in the concentration of 0.05 microgram/ml disturbs a correlation between the morphological cycle of mitochondria and phases of the mitotic cell cycle which is characteristic of intact cells. An increase in the total number of mitochondria independent of phase is observed in all the cells in comparison with intact cells. At the same time a decrease in the amount of branched organellae and appearance of giant mitochondria are discovered. All mitochondria are in the condense form. These changes, perhaps, are a result of the inhibition of the rRNA synthesis in the nucleus and of the protein synthesis in the cell found with it by AMD. The possibility of the immediate interaction of AMD with membrane components of the cell, which induces changes in the ion concentrations and peroxidation of the membrane lipids is not excluded.     

Urethane inhibition of DNA synthesis in the hepatocytes of the regenerating liver in mice

Hepatocarcinogen urethane (ethyl carbamate) inhibits DNA synthesis in the regenerating mice liver when administered at the peak of stimulated proliferation--46 hours after partial hepatectomy. The inhibition is temporary and reversible. The maximum inhibition of 3H-thymidine incorporation in the cells is observed 12 hours after urethane administration, and the effect is removed following 20 hours after administration. Another effect of urethane consists in the lengthening of the period of DNA synthesis by 1.38 times, as estimated by the Quastler-Sherman method, though it does not affect the length of G2-period or mitosis. Possible mechanisms of the effect of urethane on the initiation of DNA synthesis and on the rate of DNA replication are discussed.     

The ultrastructural basis of steroid production in the Y-organ and the mandibular organ of the crabs Hemigrapsus nudus (Dana) and Carcinus maenas L.

Summary The ultrastructure of the steroid producing Y-organ and the mandibular organ of the crustaceans Hemigrapsus nudus and Carcinus maenas has been studied with reference to the well investigated steroid secreting cells (SSC) of mammals. In accordance with the most important characteristic of mammalian SSC, abundant SER could be shown in the Y-organ, where it is unevenly distributed. The amount of SER seems to vary in correlation with the secretion of moulting hormone during the moult cycle. Most Y-organ cells contain a great number of mitochondria of the tubular type, another important characteristic of mammalian SSC. The ultrastructure of the mandibular organ of C. maenas differs considerably from that of the Y-organ. Some SER was found, mitochondria of unusual shape and size were conspicuous. No definite conclusion as to the function of the mandibular organ is yet to be drawn.Dedicated to Prof. Dr. Peter Karlson on the occasion of his 60th birthdaySupported by the Deutsche Forschungsgemeinschaft, grant Ad 24/4We wish to thank Dr. A. Owczarzak, Oregon State University, Corvallis, Oregon, for providing the facilities for our work with H. nudus and Thomas Gallenstein for many helpful discussions of technical problems     

Cell population kinetics and the cell population mechanisms of the circadian rhythm of proliferative activity in mouse esophageal epithelium]

The dynamics of 3H-thymidine labeled mitosis and diurnal rhythm of proliferative activity was studied. The isotope was injected to BALB/C mice at the peak of diurnal rhythm of DNA synthesis activity of basal layer cells of oesophageus epithelium. It has been established that the increase in the mitotic index during 24 hours depends on the increase in number of cells being in S-period. The data show that the increase of mitotic index at diurnal rhythm occurs at the expense of 75% of new G0-cells which entered into the mitotic cycle, and of 25% of re-entering cells that had divided during the maximal mitotic activity a day before. It is found that the duration of mitotic cycle of cell population which entered into the mitotic cycle synchronously is almost equal to the period of diurnal rhythm of mitotic activity, i.e. 24 hours.     

Rate of progress of the 1st post-stimulation division of the mitotic cycle by cells of ascitic hepatoma 22A of different ages]

A study was made of the progress rate of cells of the ascitic hepatoma 22A of different age during the iirst mitotic cycle after the stimulation of division. The "ageing" (11-day), terminal (14-day), and "delayed" (4 days older than the terminal stage) ascitic fluids were used. The maximal values of the labeled nuclei index was found to be reached by 9--12 hours (it was mainly due to the transtion of the quiescent to the S-period) and the maximal mitotic index--by 18--21 hours after the inoculation, independently of the tumour age. These results suggest that the duration of both the prereplicative (G1) period and of the whole first mitotic cycle after the stimulation were independent of the time during which the cells of the ascitic hepatoma 22A were at the resting stage or at the very prolonged G1-period.     

Temperature acclimation in the teleost,Blennius pholis: Changes in enzyme activity and cell structure

• 1.1. The effect of thermal acclimation on the activity of liver enzymes of B. pholis and on the ultrastructure of the liver was studied.
• 2.2. Significant increases in the activities of SDH and G6PDH, but not of LDH were found after cold-acclimation.
• 3.3. E.M. measurements suggested that cold-acclimation resulted in an increase in the size of liver cells and their nuclei, and in the number, but not the size of liver mitochondria.

     

Effect of irradiation on the size of the mitochondria in rat cardiomyocytes

The paper presents the results of studies of the influence of radiation on the sizes of cardiomyocyte mitochondria in rats. Considering the established regularity of subordination of mitochondrial size distribution to the gamma-law on the cardiomyocyte mitochondria, one can judge presumably on their functional state by the variation in the mitochondrial size distribution. This conclusion is confirmed by the relationship between the redistribution of mitochondrial size and the biochemical parameters of their functional activities. It is recommended to evaluate morphometrically the variation in mitochondrial sizes in the course of computer-assisted microscopic investigation of cells for the purpose of assessment of morphological and functional properties of these cellular organellae and the cell as a whole.     

Geometric control of the cell cycle

How do cells sense their own size and shape? And how does this information regulate progression of the cell cycle? Our group, in parallel to that of Paul Nurse, have recently demonstrated that fission yeast cells use a novel geometry-sensing mechanism to couple cell length perception with entry into mitosis. These rod-shaped cells measure their own length by using a medially-placed sensor, Cdr2, that reads a protein gradient emanating from cell tips, Pom1, to control entry into mitosis. Budding yeast cells use a similar molecular sensor to delay entry into mitosis in response to defects in bud morphogenesis. Metazoan cells also modulate cell proliferation in response to their own shape by sensing tension. Here I discuss the recent results obtained for the fission yeast system and compare them to the strategies used by these other organisms to perceive their own morphology.     

Rate of DNA replication in the DNA synthetic period of the barley chromosomes

The rate of DNA replication, as judged by H³-thymidine incorporation, at the specific time of the S-period in chromosomes of barley (Hakata No. 2) is studied by means of autoradiography.In the barley chromosomes, two different DNA units with respect to replication-time are distinguishable. The early replicating DNA is replicated at least within 1 hour ab init. of the S-period, and the late replicating DNA within 1/2 to 1 hour before the end of the S-period. The replication scarcely occurs in the middle of the S-period. These evidences suggest that the replication of chromosomal DNA in the present material does, therefore, not proceed in a continuous time sequence. Topographically, the early replicating DNA is almost confined exclusively to the distal regions of the chromosomes 1 and 5, and this situation seems applicable to other chromosomes as well, whereas the late replicating DNA is close to the centromere on its both sides. Hence, the replication of chromosomal DNA does not proceed uniformly in a longitudinal sequence along the chromosomes. The interrelationships among chromosome structure in its cytological expression, replication -pattern and -time of chromosomes, and regulating mechanisms of DNA replication are discussed.