Mitosis in embryonic trisomy 1 mouse eye

Trisomy 1 embryos consistently show eye defects (e.g., aphakia, microphakia, retention of lens stalk). To determine if the plane of division of mitotic figures is abnormal in the eyes of these animals, trisomic embryos (9.5 through 12 gestational days) were produced from mice doubly heterozygous for Robertsonian translocation chromosomes [Rb(1.3)/Rb(1.10)]. To accommodate for the known delay in trisomic embryo development, animals were grouped according to stages of eye development rather than to gestational age. Serial sections were evaluated without knowledge of karyotype for orientation of mitotic figures (parallel, perpendicular, oblique) in lens, optic cup, and diencephalon. Location of mitotic figures was scored as apical (nearest the lumen), middle, and basal. Numerous anomalies were noted in trisomic eye development. No difference was found between orientation of mitotic figures in the lens and optic cup of trisomy 1 and normal embryos. Location of mitotic figures in trisomy 1 lens was significantly different from that of normal littermates. The data confirm earlier studies that trisomy 1 affects the eye, and they tend to corroborate evidence that this trisomy affects the lens more than it affects the optic cup.     

Effects of methylmercury on the morphogenesis of the rat cerebellum

Developing rat cerebellums were examined following continuous methylmercury exposure via maternal drinking water at 12.5 ppm during gestation and the suckling period. The continuous exposure resulted in reductions of the total cerebellar cell population and higher mercury tissue burdens than previous acute-dose studies. Cell necrosis was not evident, but rather alterations in the pattern of mitotic figures were observed. A decreased number of cells in the methylmercury exposed cerebellums was associated with an increased number of mitotic figures in the early stages of mitosis and a decrease in the number in the middle and late stages. These in vivo exposure observations are consistent with in vitro cell cycle studies in which the cells were found to have accumulated in G2 and early M phases. Impaired cell proliferation is suggested to be a major mechanism of developmental neurotoxicity following continuous low-dose exposure to methylmercury.     

Occurrence and possible consequences of multipolar mitoses in primary cultures of adult rat hepatocytes

Proliferating primary cultures of adult rat hepatocytes are characterized by the occurrence of multipolar mitoses, and chromosome loss resulting in the formation of micronuclei at telophase. The percentage of multipolar mitotic figures was determined to be 12.76 ± 7.9%, 80% of which were tripolar. Multipolar mitotic stages showed a high incidence of chromosome loss, increasing from meta- (61.7 ± 16.6%) to telophase (72.1 ± 19.3%). Regular bipolar mitotic figures on the other hand also showed chromosome loss, however, to a lesser degree and decreasing from meta- (49.5 ± 10.4%) to telophase (34.9 ± 7.9%). The incidence of chromosome loss even in regular mitotic figures is very high compared to other cells and appears to depend on another special feature of hepatocytes: they remain flat and well attached during mitosis, so that shearing forces could be responsible for the separation of chromosomes from the mitotic spindle. Additionally this morphology creates a situation allowing for a maximal interaction of mitotic spindles of binucleated cells, leading to the high rate of multipolar mitoses observed. Both multipolar mitoses and chromosome loss could also explain the consecutive detachment of hepatocytes reported for proliferating primary cultures, since the aneuploid daughter cells generated can be expected to be nonviable in most cases and eventually detach. © 1993 Wiley-Liss, Inc.     

A selective stain for mitotic figures, particularly in the developing brain

A selective stain for mitotic figures is valuable where autoradiographic counting is not required, especially in the developing brain. Most work in this field has been based on conventional nuclear stains which do not differentiate mitotic figures from resting cells by color. Hematoxylin, Feulgen, gallocyanin and Nissl methods have been used particularly. The method described uses a modified Bouin fixative, followed by hydrolysis in 1 N HCl. Mitotic figures are selectively stained using crystal violet, with nuclear fast red as the counterstain for resting cells. The method has been tested using material from postnatal and fetal sheep, guinea pig and rat. Using paraffin mounted serial sections it is applicable to all organs. The method was very successful on developing rat brain, particularly for detail and quantitative estimation in the early stages of prenatal development, which was of primary interest. Nucleated cells of the erythrocytic series, keratin and what appear to be mast cells were found to stain. When nuclear counting or cell recognition were required these did not cause any difficulty, except in prenatal liver. The highly selective method presented stains mitotic figures, in all tissue tested, an intense blue against a background of red resting cells.     

A Selective Stain for Mitotic Figures, Particularly in the Developing Brain

A selective stain for mitotic figures is valuable where autoradiographic counting is not required, especially in the developing brain. Most work in this field has been based on conventional nuclear stains which do not differentiate mitotic figures from resting cells by color. Hematoxylin, Feulgen, gallocyanin and Nissl methods have been used particularly. The method described uses a modified Bouin fixative, followed by hydrolysis in 1 N HCl. Mitotic figures are selectively stained using crystal violet, with nuclear fast red as the counterstain for resting cells. The method has been tested using material from postnatal and fetal sheep, guinea pig and rat. Using paraffin mounted serial sections it is applicable to all organs. The method was very successful on developing rat brain, particularly for detail and quantitative estimation in the early stages of prenatal development, which was of primary interest. Nucleated cells of the erythrocytic series, keratin and what appear to be mast cells were found to stain. When nuclear counting or cell recognition were required these did not cause any difficulty, except in prenatal liver. The highly selective method presented stains mitotic figures, in all tissue tested, an intense blue against a background of red resting cells.     

Proliferation of glial cells in vivo induced in the neural lobe of the rat pituitary by lithium

Lithium salts are widely used for treatment of psychiatric illness. Lithium also affects cell proliferation. During investigation of the effect of lithium chloride on the central nervous system (CNS) of nephrectomized rats, we noted numerous mitotic figures in the neural lobe of the pituitary. Morphologic criteria established that the mitotic cells were astrocytes, the supporting glial cells of the CNS, also known as pituicytes. Equimolar doses of chlorides of chemically related cations (sodium, potassium, rubidium) had no such effect.     

Structure and Cytogenesis of the Shoot Apex of Syringa oblata var. affinis Lingelsh

The structure, growth and mitotic activity of 211 shoot apices of developing sprouts of Syringa oblata var. affinis Lingelsh. in longitudinal sections and 67 in transverse sections have been studied with the view to understanding the nature of zonation patterns and cytogenesis of the apical meristems during a double plastochron. The external morphology and the anatomical structure of the apices in 4 plastochronic stage-early, middle, late Ⅰ and late Ⅱ stages are described. In the shoot apices examined, especially those at late plastochronic stage, the following zones may be delimited: Zone of tunica initials, zone of corpus initials, peripheral zone and zone of rib meristem. The location and orientation of mitotic figures observed in longisections of the apices in 4 plastochronic stages are plotted in diagrams and the mitotic frequency has been calculated. Information obtained from these investigations reveals that the tunica and corpus inititals constitute an active region of the apex, but their mitotic activity changes periodically within the double plastochron. In the middle plastochronic stage when the apex is at its minimal area and the cells of peripheral zone and rib meristem zone have been completely transformed into constituent parts of foliar primordia and the subjacent tissues of the stem and the pith mother cells respectively, the mitotic frequency of the initials is at its maximium and its intensity of mitotic activity is not much lower than that of any other meristematic zone at any stage. When the apical dome is reformed by the activity of these initials in late plastochronic stages, the mitotic frequency of the initials gradually drops and the region of high mitotic frequency shifts to the flank of the apex, the peripheral zone. Anticlinal divisions are predominant in this zone. On the other hand, those cells directly left behind by the corpus initials, which constitute the rib meristem, are vacuolated and marked by the pre- dominance of transverse divisions. Thus the entire zonation pattern reappears. In the next early plastochronic stage, the mitotic frequency of the tunica and corpus initials drops to its mimimium, but other regions of the apex still maintain a high mitotic frequency. It may be concluded that the tunica and corpus initials form a cytogenerative center of the shoot, and the cytohistological zonation is actually a result of the fact that different regions of apical meristems are different in mitotic activety, different in state of cell differentiation and different in their function in morphogenesis.     

Quantitative and qualitative characteristics of the stages and transitions in the cycle of the rat seminiferous epithelium: light microscopic observations of perfusion-fixed and plastic-embedded testes

The stages of the cycle in the rat seminiferous epithelium are illustrated for testes fixed by vascular perfusion and embedded in plastic resins. Improved cellular resolution in plastic sections permitted a clearer demarcation of the stages than in paraffin. Quantitative data are presented to support the recognition of stages, particularly those in transition. Stages IV, V, VII, XI, and XII had the highest frequencies of transitional characteristics. Stage IV was redefined to be more consistent with the occurrence of a high percentage of mitotic figures and to clarify transitions in this stage. Although the resolution of cellular detail was greatly improved with the use of plastics, the thinner sections contained fewer identifying features together within a single tubule cross section and sometimes major characteristics were absent. Therefore, additional characteristics were used for stage classification, such as nuclear diameter and the presence or absence of mitotic figures. A binary decision key is provided to improve consistency among laboratories in the identification of the stages in plastic-embedded testes.     

A new action for topoisomerase inhibitors.

Topoisomerases are known to aid DNA replication by breaking and resealing supercoiled DNA. Consequently, cells exposed to topoisomerase inhibitors before or during the S (DNA synthetic) phase of the cell cycle undergo abnormal DNA replication and become irreversibly blocked in the G2 (pre-mitosis) phase. We report that following a 4-h exposure to topoisomerase II inhibitors, murine erythroleukemic cells (MELC) do not form mitotic figures but exhibit a time-dependent progression into G2 (4N DNA) and greater than G2 (up to 8N DNA) stages of the cell cycle. Following exposure to the topoisomerase I inhibitor camptothecin, recovering MELC also exhibit greater than G2 polyploidy, but to a considerably lesser degree: mitotic figures are present and a subpopulation of cells resumes cycling. However, both topo I and topo II inhibitors induce maximal percentages of greater than G2 cells when synchronized MELC are in the G2/M phase at the time of exposure. This suggests that, in addition to their S-phase action, topoisomerase inhibitors can interfere with chromosome condensation during G2 and, in so doing, induce polyploidy.     

Sustained Isoprostane E2 Elevation,Inflammation and Fibrosis after Acute Ischaemia-Reperfusion Injury Are Reduced by Pregnane X Receptor Activation

Liver grafts donated after cardiac death are increasingly used to expand the donor pool but are prone to ischaemic-type biliary lesions. The anti-inflammatory effects of the activated pregnane X receptor have previously been shown to be beneficial in a number of inflammatory liver conditions. However, its role in reducing peri-portal inflammation and fibrosis following ischaemia-reperfusion injury has not been investigated. Hepatic injury and its response to pregnane X receptor activation was examined after partial hepatic ischaemia-reperfusion injury induced by surgically clamping the left and middle lobar blood vessels in rats. Molecular and pathological changes in the liver were examined over the following 28 days. Ischaemia-reperfusion injury resulted in transient cholestasis associated with microvillar changes in biliary epithelial cell membranes and hepatocellular injury which resolved within days after reperfusion. However, in contrast to chemically-induced acute liver injuries, this was followed by sustained elevation in isoprostane E2, peri-portal inflammation and fibrosis that remained unresolved in the ischaemic reperfused lobe for at least 28 days after clamping. Administration of pregnenolone-16α-carbonitrile—a rodent-specific pregnane X receptor activator—resulted in significant reductions in cholestasis, hepatic injury, ischaemic lobe isoprostane E2 levels, peri-portal inflammation and fibrosis. Hepatic ischaemia-reperfusion injury therefore results in inflammatory and fibrotic changes that persist well beyond the initial ischaemic insult. Drug-mediated activation of the pregnane X receptor reduced these adverse changes in rats, suggesting that the pregnane X receptor is a viable drug target to reduce ischaemic-type biliary lesions in recipients of liver transplants donated after cardiac death.     

Characteristics of trophoblast cell reproduction in the connective zone of the rat placenta. II. Determination of the degree of ploidy of mitotic shapes

Morphological and cytophotometric studies have been made on polyploidization of placenta connective zone cells. Measurement of the DNA content in mitotic figures show that within a period of development ranging from day 13 to day 14 the bulk of mitoses (up to 25%) become tetraploid and octaploid. This may suggest that polyploidization of placenta connective zone cells proceeds via incomplete polyploidizing mitoses. Among tetraploid and octaploid mitotic figures, there are those corresponding to all the mitotic stages, from prophase to telophase. Consequently, mitosis in tetraploid and octaploid cells can reach telophase. In such cases polyploidization is likely to follow the acytokinetic mitotic pattern. A question of a certain maximum level of polyploidy that may be reached by cells due to the incomplete mitosis is discussed.     

白鲟肝脏和胰脏的组织学与形态学研究

白鲟肝脏较大,可分为左右两大叶及小的中叶,胆囊位于右叶的凹缺内。肝板多由双层细胞构成,肝小叶不明显。肝内毛细胆小管由4个肝实质细胞围成。胰脏有3支,被厚的浆膜。胰岛明显。胰管与胆管汇合后共同开口于小肠最前部背面。对肝、胰实质细胞的显微或亚显微结构进行了描述。     

Development of the quiescent center in maturing embryonic radicles of pea (Pisum sativum L. cv. Alaska)

Maturing embryos of pea (Pisum sativum L. cv. Alaska) were treated with an aqueous solution of tritiated thymidine for 1 h, sectioned, and processed for autoradiography. An analysis of the distribution of labelled nuclei and mitotic figures demonstrated the presence of a quiescent center (QC) in the radicles of developing embryos. The QC developed in the radicle during the growth of the embryo. Immature radicles that did not contain a well-formed zone of root-cap initials did not show a QC. In the latter stages of seed ripening, the pattern of arrest of DNA synthesis and mitosis was tissue-specific. Cells within the QC remained inactive. The region lacking labelled nuclei and mitotic figures progressively expanded to include the root cap initials and then the provascular cylinder. Mitosis was arrested before DNA synthesis in the embryonic cortex. Cells within the QC synthesized DNA during the first stages of seed germination.Abbreviations [³H]TdR tritiated thymidine - QC quiescent center     

Electron Microscope Studies of Nuclear Changes in Saccharomyces Cerevisiae During Bud Formation

Ultraviolet photolysis was used to obtain transparent yeast cells when observed with the electron microscope. Following proper irradiation the yeast nucleus was easily discernable and appeared to be in various stages of division. Spindle-like figures were seen which give weight to the concept of a mitotic process for the division of the yeast nucleus. Distinct chromosomes were not observed.     

Hepatic carcinoma with spleen metastasis in a California sea lion from the Gulf of California.

A primary hepatic carcinoma with a neuroendocrine pattern was detected in an adult female California sea lion (Zalophus californianus) found dead on Granito Island in the Gulf of California (Mexico) in January 1996. At necropsy, several light yellow nodules of different sizes were observed on the entire surface of the liver and spleen. Microscopic examination of these nodules using routine haematoxylin-eosin stain, revealed cubic, polyhedral and pleomorphic cells with three to four bizarre mitotic figures per field (40X). An immunohistochemistry test revealed a positive reaction of indirect immunoperoxide to cytokeratin (CK2). This is the first known case of a primary hepatic carcinoma in free-ranging California sea lions from Mexican waters.     

Structure of chromatin and chromosomes in preparations of interphase nucleus derivatives, prepared by removal of the nucleuar envelopes. II. Structure of chromatin and associations of chromosomes in stretched amembranous nuclei and mitotic figures]

Preparations of surface stretched amembranous nuclei and mitotic figures were used for revealing the high order nuclear and chromosomal structures. The preparations were obtained by dropping amembraneous nuclei and mitotic figures suspension in methanol-glacial acetic acid mixture (3:1) on wetted superclean slides. Amembraneous nuclei and mitotic figures were isolated from intact murine and human cells (lines L1210, SK-UT-1B, PHA-stimulated lymphocytes) by means of their 1-5 min prefixational capillary pipetting with freshly prepared 0.018-0.06% Triton X-100 solution in the conditional cultural medium. Stretched amembraneous nuclei and mitotic figures had no features of induced chromatin dispersion and compaction. Stretched interphase amembraneous nuclei showed spatially separated individual structures (thin chromatin fibres, nucleoli, intranuclear bodies), polymorphous pattern of perinucleolar chromatin aggregation and episodically expressed beaded thick chromatin fibres and a chromocenter. The chromomeric pattern of the spread chromosomes of mitotic figures was quite similar but hardly identical with that of G-banding. The stretched prometaphase mitotic figures in all tested cell types always contained loose "residual" nucleoli looking like typical prophase nucleoli as concerns their shape and number per cell (mitotic figure). The majority of chromosomes of stretched mitotic figures and of prophase amembraneous nuclei were attached to the nucleolar material. All tested cell lines showed almost the same variation in number of nucleolus-attached chromosomes, per both prophase amembraneous nucleus and prometaphase mitotic figure. Some chromosomes of stretched mitotic figures were colocated with "residual" nucleoli and looked shortened and strongly condensed. Other chromosomes, locally associated with "residual" nucleoli, were straight and oriented radially to these. Mutual chromosomal arrangements in mitotic cells on smears and in stretched mitotic figures were analogous. Equatorial plates from PBS-washed SK-UT-1B cells displayed a better stretching capacity than those from untreated cells. In the former case metaphase chromosomes were seen more uniformly stretched and well identified after GTG-banding procedure. The number of interchromosomal (mainly telomere-telomeric and telomere-centromeric) connections per stretched mitotic figure (or per stretched prophase amembraneous nucleus) was minimum in late prometaphase, maximum in prophase and early prometaphase, and intermediate in metaphase. The obtained data are discussed in terms of topology and longitudinal heterogeneity of mitotic chromosomes.     

Regional differences in mitotic activity due to injury in mouse skin

Summary Mitotic activity adjacent to a wound inflicted at different sites in the mouse skin was measured 24 h after injury. A regional difference in the epidermal mitotic activity due to injury was noted. Mitotic activity was high in the anterior parts of the body including the head, lower in the middle to posterior regions of the body and lowest in the posterior-most parts of the body. Regional differences in epidermal mitotic activity due to injury were demonstrated in both female and male mice. The existence of a cranio-caudal gradient in epidermal response to injury is suggested.     

Indomethacin inhibits cell proliferation in the oxyntic epithelium of the rat

The aim of this investigation was to examine the action of parenteral indomethacin and oral prostaglandin E₂ on cell proliferation in the rat oxyntic mucosa. Groups of Sprague Dawley rats were treated with either 1.5 mg/kg indomethacin subcutaneously, 5 mg/kg oral prostaglandin E₂ or placebo, twice daily during 5 days. All rats were killed exactly 4 hours after mitotic arrest with vincristine, and a biopsy specimen from the oxyntic mucosa was processed for routine microscopic evaluation.Mitotic figures were distributed cluster-like along the oxyntic mucosa alternating with mitosis-free areas. The total number of mitotic figures in 8 mm of mucosa was significantly reduced by administration of indomethacin (p<0.05). In rats given indomethacin, 32.5% of the examined mucosa did not have mitotic figures, which is significantly higher than 14.3% as observed in placebo-treated rats (p<0.05). Both rats treated with indomethacin and with prostaglandin E₂ had fewer microscopic fields containing 5–6 mitotic figures than placebo-treated animals (p<0.05). The maximal length of mitosis-free areas was 0.6 (0.6–0.9) mm in rats given indomethacin which is significantly larger than 0.4 (0.2–0.4) mm observed in controls (p<0.05). Indomethacin produced epithelial atrophy as shown by a significant reduction of the epithelial height observed in those rats compared to controls (p<0.05).The inhibition of cell proliferation observed in the oxyntic mucosa of rats treated with the cyclooxygenase blocker indicates that an important physiological role of endogenous prostaglandin is to maintain the proliferative activity of the epithelium at a high level.     

Effect of Thioacetamide on Rat Liver Regeneration : II. Nuclear RNA in Mitosis

The effect of thioacetamide on dividing cells of regenerating rat liver has been studied. Rats were given daily subcutaneous injections of thioacetamide as a 1 per cent solution at a dosage of 5 mg./100 gm. body weight for 7 to 10 days, subjected to partial hepatectomy, and sacrificed 28 to 31 hours later. Thioacetamide treatment results in striking increases in the nuclear ribonucleoproteins of the liver cell without affecting the mitotic rate during regeneration (14). During mitosis, RNA-containing particles were seen within the spindle and coating the contracted chromosomes from prophase through metaphase or early anaphase. At telophase, prior to the reconstruction of the nuclear membrane, fine RNA-containing granules appeared within the compact chromosomal groups. These coalesced to form nucleoli corresponding in number to the number of nucleolar organizer regions. The nuclei and nucleoli showed a rapid increase in size during the reconstruction period when compared with corresponding figures of the control liver samples. Electron micrographs of interphase nucleoli indicated a similar basic granular structure in both drug-treated and control animals. The question is raised as to whether the increased nucleolar material merely made visible some of the nucleolar-chromosomal associations that normally occur in mitosis, or whether thioacetamide directly affects the synthetic activity of the contracted mitotic chromosomes.     

Proliferation of glial cells induced by lithium in the neural lobe of the rat pituitary is enhanced by dehydration

Rats dehydrated by 6 days of water deprivation had a low level of mitotic activity in the astrocytes ('pituicytes') of the neural lobe of the pituitary. Mitotic activity in the pituicytes was greatly increased when isotonic lithium was administered in the last 3 days of water deprivation. Rehydration on the last day of the experiment produced a further increase in mitoses. Isotonic solutions of sodium, potassium or rubidium chloride did not increase mitoses. This model of cell proliferation is of interest because the mitotic activity is related to a physiological attempt to maintain homeostasis rather than a response to injury or the development of neoplasia.