Binuclear Cells Research Articles

Existence of hepatic cell classes and their relationship with the nuclear classes in the liver of the white mouse. A microinterferometric study.

1. Individual hepatocytes isolated from unfixed liver of the white mouse are different in their solid contents. However, they fall into weight groups that form geometrical or arithmetical progressions as 2∶4∶8∶16 and 1∶2∶3∶4∶..., depending on the mouse age. These weight groups are here named ‘cellular classes’. The hepatocytes of suckling and weaning mice are relatively small and form 4 cellular classes disposed in geometrical progression, whereas those of adult mice are heavier and form 10 or more cellular classes disposed in arithmetical progression. 2. Binuclear cells are present in all the cellular classes of the adult liver, whereas they contribute to constitute only the third and fourth cellular classes of the suckling mice liver. 3. The water-soluble contents of individual hepatocytes is constant as a percentage, being about 40 per cent of the total cell dry mass in both suckling and adult mice. 4. The nucleo-cytoplasmic weight ratio of hepatocytes in suckling and adult mice is constantly between 1∶2.8 and 1∶14.2. 5. The hepatocyte polyploidization occurs after the solid constituents of the cell have undergone an increase. So it might represent a mechanism to maintain the nucleo-cytoplasmic weight ratio within the range (1∶2.8)–(1∶14.2). 6. The developmental growth of the liver specific tissue takes place mainly owing to cell number increase from birth to 15 days of life, thenceforth owing to cellular hypertrophy. 7. In the early ages the hepatocyte physiologically enlarges by doublings of its contents, after the weaning time by subsequent additions of a constant amount of cytoplasm.

Relationship between diurnal rhythm in the number of binuclear cells in the rat liver and its glycogen-forming function

Relationship between diurnal rhythm in the number of binuclear cells in the rat liver and its glycogen-forming function

Effect of feeding time and lighting conditions on circadian periodicity in number of binuclear cells and mitoses in the rat liver

Effect of feeding time and lighting conditions on circadian periodicity in number of binuclear cells and mitoses in the rat liver

Stimulation of incorporation by triiodothyronine of thymidine-methyl- 3H into hepatic DNA of the rat

Administration of single dose of triiodothyronine (T 3) results in hepatic cell proliferation as judged by the mitotic index and the enhanced thymidine-methyl- 3H or orthophosphate- 32P incorporation into DNA. Increased mitoses could be observed 24 hours after T 3 administration; 72 hours after the hormone injection, binuclear cells were abundant. The enhanced hepatic DNA synthesis reached its peak 24–48 hours after T 3 injection. Radioautographic techniques revealed that T 3 increases DNA synthesis primarily in the hepatic cells. T 3 had no effect on testicular DNA synthesis. T 3-induced liver DNA synthesis was sensitive to the inhibitory effects of actinomycin D and cycloheximide. The present observations indicate that the early effects of thyroid hormone on liver protein and RNA synthesis initiates DNA synthesis, which in turn leads to the increased proliferation of hepatic tissue.

Polyploïdie somatique dans le foie de rat: I. Le rôle des cellules binucléées dans la genèse des cellules polyploïdes

The formation of polyploid cells in the rat liver was studied in relation to the variation of the number of binuclear cells, directly determined by means of the method of liver cell suspensions. The polyploid cells appear in large numbers during the various types of growth of the hepatic tissue. Then, their appearance is strictly linked with the disappearance of a corresponding number of preexisting binuclear cells. This mechanism was verified in the course of the normal physiological growth and after partial hepatectomy, carried out on animals of various ages. It is impossible to induce the formation of polyploid cells in a liver which does not contain any binuclear cells (partial hepatectomy on animals less than 3 weeks old). These results clearly show that the binuclear phase is an intermediate stage between a given state of polyploidy and the immediately superior one. The binuclear cells give rise to two cells of higher ploidy owing to a particular kind of mitotic division. After a simultaneous prophase in both nuclei and the formation of only one spindle, the chromosomic groups fuse, giving a dipolar anaphase with a double chromosomic arrangement. Thus, the problem put by the hepatic polysomatism is mainly a question of determinism and mode of formation of binuclear cells.

Polyploïdie dans le foie de rat: II. Le rôle de l'hypophyse et de la carence protéique

Polyploidy occurs through a series of cellular divisions of particular types, the first of which is the formation of binuclear cells. Therefore the liver of an animal must be in a state of growth for polyploid cells to be formed. It is possible to prevent polyploid hepatocytes from appearing by bringing about in young animals either a hypophysis insufficiency or a protein deficiency large enough to stop the development of the animal. The lack of growth inhibits the formation of polyploid cells even if the growth hormone is given by injection. The factors inducing the formation of polyploid cells should therefore be tested by their action on hepatic cell multiplication in baby rats (three weeks old or less) which normally show very few binuclear or polyploid cells.

DNA content of the corneal epithelial cells of rats after x-irradiation

In a period of 24–120 h after the irradiation of rats with x-rays in a dose of 700R there is a marked nuclear polymorphism. In the corneal epithelium there appear cells with satellite nuclei and a larger number of binuclear cells. The results of investigations carried out by means of cytospectrophotometry showed that after irradiation the number of polyploid nuclei containing DNA in quantities of 5–8 respectively grows 30% whereas normally it is only 10%. The satellite nuclei contain an insiginficant amount of DNA, which points to their formation from chromosome fragments during atypical karyokinesis. The amount of DNA in the nuclei of binuclear cells varies extensively; the minimum quantity being 1n+1n, the maximum we have noted being 3n+5n.

The diurnal rhythm of mitotic activity in the regenerating liver during acute radiation sickness

In experiments on albino rats a study was made of the 24 h thythm in the mitotic activity of hepatic cells and in the percentage of binuclear cells in the regenerating liver following a single total exposure to X-irradiation in a dose of 600 r. The finding was that at the peak of acute radiation sickness the 24 h mitotic rhythm was retained. As in the case with reparative and physiological regeneration in nonirradiated animals the peak of mitotic activity occured during the early morning hours. The mean 24 h coefficient of the cell mitotic activity (2–3 days following operation and correspondingly 5–6 days following irradiation) did not diminish as compared to control. No regularities were revealed that would suggest that during the 24 h period the number of binuclear cells in irradiated and nonirradiated rats was changed.

Binuclear cells and proliferative processes in the intact and regenerative liver of rats

In the nonoperated liver of control female rats 2 to 17 months after the beginning of the experiment there was a gradual increase of the number of binuclear cells and their sizes; the size of the nucleus remained unchanged. 2–5 months after the removal of two thirds of the liver the percentage of binuclear cells was higher than in the liver of the control animals of the same age; moreover hypertrophy of binuclear cells and their nuclei was observed at these periods. At the late postoperative dates (in 17 months) the percentage of binuclear hepatic cells decreased; their size and the size of their nuclei also became smaller.

Some Mechanisms of Formation of Polyploid and Heteroploid Cells in a Murine Ascites Tumor <italic>In Vitro</italic>

This paper records the behavior in vitro of cells, at and about the time of their division, in an ascites subline of a naturally occurring murine epithelioma. Under the conditions of these experiments, uninuclear cells of higher ploidy are formed from uninuclear cells of lower ploidy through the intermediate stage of a binuclear cell. The production of the intermediate binuclear cell depends on failure of cytoplasmic cleavage at the time of attempted division of a uninuclear cell. When a binuclear cell divides to produce 2 uninuclear daughters, the chromosomes of both nuclei come together on a single metaphase plate, anaphase and telophase appearing normal. Cytoplasmic cleavage is successful, and two uninuclear daughters are formed. Uninuclear daughters of higher ploidy so produced are viable and capable of further mitoses, which may be normal or may vary in the modes of their division. In a binuclear cell that fails to cleave in successive attempts at division, the chromosomes come together on a single metaphase plate on each occasion, which results in a cell of successively higher ploidy. Rarely, multipolar separation of chromosomes results in reduction of the chromosome complement of a cell. Similar multipolar divisions may give rise to binuclear and trinuclear cells that are usually viable and may undergo further division.

Beobachtung der Bildung zwei zweikerniger Zellen aus einer Zelle der Gewebekultur ohne experimentellen Eingriff

The author reports on the development of two binuclear cells from one PK cell dividing mitotically, which, by its aberrant chromosomes, recalls tripolar mitosis. He believes that this process is not an exception and that it may be used for the explanation of the development of a certain part of binuclear cells frequently found in stabilized strains.

Ratio between the numbers of polyploid and binuclear liver cells during the 24-hour period

Experiments were staged on male albino rats weighing 150 gms. Animals were sacrificed at 11, 2, 5, 8, 11, 1 and 8 o'clock. Pieces of the left lobe of the liver were obtained for the examination. The number of binuclear and of the large polyploid (octoploid) cells were counted: the number of mitoses and destroying nuclei and their correlation at different time of the day was also estimated.

Transfert nucleaire intercellulaire en cultures mixtes in vitro

Following previous publications [1, 2, 3, 4] concerning the appearance in associated in vitro cultures of two different cell strains of a new type of cells having “hybrid” characteristics, we have investigated the cytological mechanisms possibly involved in this phenomenon. With this aim, explants of mouse normal embryonic tissue were associated in plasma clot cultures with aggregates of cells from a highly malignant mouse cell strain (N1 derived from the NCTC 2472 Sanford clone). The areas of contact between the respective growths were observed with the aid of phase contrast, time-lapse microcinematography. Two cases of transfer of nucleus from one cell to another in this area were checked frame after frame and the movements of the involved cells and their organellas analyzed. In the first case a penetration of an interphasic nucleus into the cytoplasm of another neighbor cell was seen: the receptor cell was most probably in a blockad early metaphase stage. The momentary constriction of the nucleus and the shift of the nucleolus through its constricted center supposes an active participation of the nucleus in this process accomplished essentially in 15 sec. In the second case, the nuclear transfer occurred between two interphase cells: the receptor cell could be identified as belonging to the N1 malignant strain. During 30 min, the penetrating nucleus advanced and turned actively inside the cytoplasm of the receptor cell. The resulting bi-nuclear cell seemed to be viable. Our observations complete some previous reports concerning intercellular transfers of cytoplasm and of its organellas. They prove that the nucleus can also participate in such intercellular exchanges which may visibly occur, as demonstrated, between cells of different histological origin as well as between normal and malignant cells.

Pathohistological Study on the Experimental Hyperestrogenism

After a certain period following castration groups of mature female rabbits were injected aubcutaneously at their back with 15 mg of estrogen pellet 1, 2, 3, 4 and 9 times at intervals of every 2 weeks, killed 2 weeks after the final administration and divided into 5 groups of 2 to 4 rabbits each according to total doses of 15 to 135 mg and periods of 2 to 18 weeks administered to produce the experimental hyperestrongenism with progesteron effect intercepted. Meanwhile, in order to confirm effccts of castration at each period, pathological, anatomical and histological examination were conducted chiefly on their uterus, mammary gland, pituitary body, thyroid gland, suprarenal gland, kidney and liver together with those of 5 control groups of 2 castrated animals each. Experimental results were classified roughly into two categories according to dose of estrogen and period administered. (1) In groups administered with 15 to 45 mg of estrogen pellet for 2 to 6 weeks 2 weeks after castration the endometrium atrophined and thinned by castration indicated a remarkable glandular proliferation, and the muscularis mucosae thinned was also hyperthophied several times. Until 6 weeks after castration the mammary gland not different from non-functional and normal control proliferated like fibroadenia and acinar adenosis, and the abmornally proliferated focus of galactophorous glands was noted. Up to 6 weeks after castration appearance of acidophile celles instead of basophile cells became remarkable in the anterior hypophysis where the latter was to proliferate, chromophobic cells also developed and eflect of castration was inhibited by administration of estrogen. The cortex became slightly hypertropeied 2 to 4 weeks after castration. Four weeks later fat decreased in the fascicular Layer of suprarenal cortex, but a large amount of fat was still encountered in the coniform layer of suprarenal cortex. However, continual decrease of fat was found in the fascicular Layer, and cells were hypertrophied from the depth to the reticular layer. The thyroid gland was reduced in function, was concentrated in colloid, and indicated large follicles due to castration. But it was weakened in the colloid, indicated small follicles and was enhanced in function due to administration of estrogen. Extending over prolonged period after castration binuclear cells developed in atrophied liver, while central vein and venous sinus were congested and enlarged. Estrogen had no effect on the spleen where there was noted no remarkable cnange after castration. Degenerated epithelium of the principal part of the nephritic duct, which became distinct 2 to 4 weeks after castration, was recoverd by administration of estrogen. (2) In groups administered with 60 to 135 mg of estrogen pellet for 8 to 18 weeks after castration the endometrium was markedly infiltrated with leukocytes showing the necroosis or the necrobiosis. The infiltra tion extended into the muscularis mucosae, and vein and cepillara of the uterine vagina were markedly congested and enlarged, part of them indicating the thrombosis. However, the residual muscularis mucosae was rather proliferative, and in groups administered with 135 mg of the medicine for 8 weeks the fibroma was formed below the serous membrane. In the mammary glands where rami of the galactophorous ducts temporarily exteded and the epithelium proliferated, lobules collapsed completely and large cystic mastopathy seemed to appear. In the anterior hypophysis surrounded by the stroma acidophile cells decreased and collapsed, while basophile cells diminished markedly and chromophobic cells increased remarkably instead. Fat in the coniform layer of the suprarenal cortex disappeared entirely, but a considerable amount of fat was noted in the fascicular layer. Concentration, collapse and disappearance of cytoblasts were noted.

A STUDY OF EXPERIMENTAL INTOXICATION OF ANTIEPILEPTIC DRUGS (PHENOBARBITAL, 5-5-DIPHENYL-HYDANTOIN, TRIDIONE)

Summary Three kinds of anticonvulsant (Phenobarbital, Hydantoin and Tridione), were administered to 12 adult rabbits and 40 adult cats for 630 days at the longest for the purpose of experimenting the acute and chronic intoxication of those drugs. 1 Concerning these three kinds, histopathological findings were almost similar. The vulnerabilities, however, were diverse as is shown in Table 2. 2 The duration factor of medication influenced much more on histopathological findings than the dosage factor. 3 The animals subjected to chronic intoxication of those drugs showed abstinent symptoms (grand ma1 seizure, vomiting, coma etc.) when administrations were abruptly cut off. 4 The changes of nerve cells are classified in following five patterns; 1) Nuclear swelling, 2) Chromatolysis, 3) Vacuolization, 4) Atrophic change and 5) Eosinophilic cell change. Furthermore, binuclear nerve cells were not seldom observed. 5 In truncus cerebri and medulla spinalis, the motor nuclei tend to show tumescent change i.e., vacuolization and chromatolysis, while the sensory nuclei presented tendency for atrophic sclerosis. 6 The author, comparing so-called ischemic cell change with other eosinophilic cell change and transitional type between these two, came to the conclusion that all of them are to be classified to eosinophilic cell changes as a whole, as there is hardly strict category of differentiation. 7 Oligodendroglial change was classified in 3 forms, namely, rosary oligodendrogliosis racemose oligodendrogliosis and so-called acute swelling. 8 Macrogliosis around the blood vessels, ventricle walls and cortical surface were classified in three types according to the arrangement of the glia cells. 9 In this experiment series also, the various histopathological findings described in the human epileptic brain, namely, so-called ischemic cell change and Chaslin's gliosis etc. were observable, however, neither Ammon's horn sclerosis nor “Prellungs Herd” were observed. 10 The author discussed the difference of vulnerability of Ammon's horn between the human and animals, trying to explain it in terms of genetic anatomy.

Innervation of Uterus in Human Embryo

The plexus cervicalis uteri shows a very good development in the medial stage of the human embryonic life, and ganglia of varied sizes are formed therein, as in the human adult. The ganglia are composed of nerve cells and paraganglion cells and are represented as my so-called compound ganglia. The connective tissue and the accessory cell-plasmodium around the nerve cells are as yet very backward in development, which presumably develop by proliferation later on accompanying the development of the nerve cells, especially their nerve processes.Nerve cells are not found in the muscular lager of the uterus.Nerve cells in the plexus cervicalis uteri are extremely well developed, as those in the other ganglia related with the urogenital organs in the minor pelvis generally are. They are classifiable into larger and smaller ones, are multipolar and can be divided into DOGIEL's types I and II by the arrangement of their nerve processes. Type I cells mostly belong to the larger and the type II cells to the smaller cells. Binuclear cells are not rare, but more multinuclear cells are not yet in existence.The paraganglia in plexus cervicalis uteri are very well developed sympathetic paraganglia, and the gland cells are each provided with a round nucleus and a cell body very hard to stain. They are rich in small veins and capillaries and are penetrated by minute nerve fibres, which are much finer than those in the ZUCKERKANDL's paraganglia observable in the latter stage of embryonic life (SAKURAOKA) and showing little change of size, pass over into the specific terminalreticulum after frequent ramifications and anastomoses, coming in tactile control over the paraganglion cells.The muscle layer of the uterus of a mid-stage embryo is composed of three distinct layers as in adult uterus, but the uterine glands are as yet immature and the connective tissue cells in the propria also infantile. Many nerve dundles, large and small, enter from the plexus cervicalis uteri into the muscular layer through the adventitia, branch out into more numerous rami and distribute widely in the tunica muscularis and the propria.Most of the fibres are fine vegetative, but a very small number of sensory fibres is found among them. The myelinisation of the sensory fibres is not yet clearly demonstrable at this stage. The perivascular plexus is also well developed, but it is of course entirely of vegetative nature.The termination of the vegetative nerve fibres is represented as the nearly completed terminalreticulum, especially in the tunica muscularis and the propria. The terminalreticulum is sometimes found running along the epithelium and the uterine glands but it never penetrate into thier cell bodies. The terminalreticulum in the vascularr wall penetrates as far as the inside of the media, but I could not observe the protoplasmatic connection between the muscle fibres and the terminalreticulum described by KNOCHE.The sensory fibres run into the connective tissue of the propria as far as the close vicinity of the epithelium, and end in sharp simplest unbranched or simple branched terminations with two to three branches. Such sensory fibres are most abundant in the cervix uteri, decreasing in the corpus and fundus, becoming very rare in the portio vaginalis.

Observations on the formation of bi-nuclear cells

In a previous communication the changes observed during the growth and division of normal cells growing in vitro were described. The present communication deals with certain modifications of the normal processes of mitosis resulting in the formation of cells containing two nuclei. Bi-nucleate cells are frequently seen in cultures of tissues grown in vitro and are generally supposed to be formed by amitotic division of the nucleus. They can be produced experimentally by the use of a slightly unfavourable medium or by subjecting the culture to the influence of some agent such as X-Rays.

Societies and Academies

LONDON. Royal Society, February 21.—C. Tate Regan: The morphology of a rare oceanic fish, Stylophorus chordatus Shaw; based on specimens collected in the Atlantic by the Dana expeditions, 1920-22. Stylophorus differs considerably in structure from its nearest allies, Trachypterus and Velifer. The skeletal differences are interpreted as related to the low elongate form, the large telescopic eyes, and the extremely protractile mouth. The unique mechanism of the protractile mouth is compared with that of the Labroid fish Epibulus.—F. P. Slater: A sensitive method for observing changes of electrical conductivity in single hygroscopic fibres. A simple electroscopic arrangement is capable of detecting the effect of small and controlled changes in relative humidity and temperature on i cm. length of a single hygroscopic fibre. The measuring system and specimen being tested are embodied in the same case, are always under the same controlled external conditions of temperature and relative humidity, and are reproducible.—T. S. P. Strangeways: Observations on the formation of bi-nuclear cells. The formation of bi-nuclear cells after mitotic division of the nucleus has been watched on living cells growing in vitro. Three types of change in the cytoplasm associated with the production of such cells are described.—J. A. Crowther: Some considerations relative to the action of X-rays on tissue cells. An analysis of the observations of Strangeways and Oakley on the immediate effects of soft X-rays on tissue cells suggests that the effect follows a probability law. This probability is the probability that ionisation will be produced in some particular body within the cell by the action of the rays. The size which this body must have to account for the observations is approximately 1/2500 mm. This appears to be of the order of magnitude of the centrosome in the cells employed in the experiments.