Calcium influence on the dry mass content and surface area of nuclei and cytoplasm during differentiation of cortex cells in pea (Pisum sativum L.) roots treated with heavy metals

The effect of Ca addition to Cd, Cr and Pb solutions on the nuclear and cytoplasmic dry mass content and its concentration as well as on these organelles dimensions were studied in cortex cells of pea roots. Ca alone, at the concentration 10(-8)M brought about a decrease (in comparison to water) in the dry mass content of nuclei and its concentration, but the increment was almost twice in the dry mass content of cytoplasm; however, it has no significant effect on its concentration. Ca ions addition does not change the surface area of nuclei except the 1st and 5th mm segments but causes a doubling of the area occupied by cytoplasm. In response to Ca addition to Cd or Cr solutions a further diminution of nuclear dry mass content takes place. Only in the case of Pb nuclear dry mass increases in the 7th mm or is similar in remaining root segments. The diminution of nuclear dry mass content due to Ca presence in metal solutions is accompanied by a lowering in its concentration, although in the presence of Cd and Pb the diminution is not significant. Ca ions addition results in an increase in cytoplasmic dry mass content. No such regularities were observed in the 1st (Cd, Pb) and 3rd (Cr) root segments. In response to Ca ions the concentration of cytoplasmic dry mass content increased insignificantly in differentiation zone and underwent reducation in the meristematic zone--in the 1st mm (Cr) and 3rd (Cd). After Ca addition to studied metal solutions the decrease in nuclear dimensions was visible only in Cd or Pb treated cells in the 3rd and 7th or in the 1st mm, respectively. An increase in nuclear size occurred only in Cr treated cells in the 7th mm. Enrichment of heavy metals with Ca caused the marked enlargement in cytoplasmic area in differentiation zone but the increment in it in meristematic zone was observed only in Cd (1st, 3rd) and Cr (1st) treated cells.     

Heavy metal action on the dry mass content and surface area of nuclei and cytoplasm during differentiation of cortex cells in pea (Pisum sativum L.) roots

The cortex cells of pea roots (Pisum sativum L.) grown for 144 h in the presence of cadmium, chromium and lead at the concentration 10(-4) M were the object of the present studies. Applied metals reduced dry mass content and concentration of nuclei in meristematic and differentiation zones. Chromium only enhanced nuclear mass concentration in the differentiation zone. The metals also made dry mass content and concentration of cytoplasm reduce, but they diminished mostly the concentration of cytoplasm in the meristematic zone and its dry mass in differentiation zone. Stimulation of cytoplasmic dry mass concentration was visible in the 1st mm (Cr) and 7th mm (Cd). Moreover, chromium caused a marked increase of cytoplasmic dry mass content in the 3rd mm of root. The studied metals reduced nuclear size, calculated as surface area, in the meristematic and differentiation zones. The increment of nuclear dimensions was observed only in the 1-3rd mm (Pb), 3rd (Cr) and 7th mm (Cd). In the presence of the applied metals the surface area of cytoplasm increased only in the 3rd mm and in 5th mm (chromium only). The present observations have shown that the toxicity of studied metals is as follows; Cd greater than Pb greater than Cr (nucleus - dry mass content and concentration, cytoplasmic area), Pb greater than Cd greater than Cr (cytoplasmic dry mass content and concentration and Cd greater than Cr greater than Pb (nuclear dimensions).     

The Growth of the Nucleus in Dividing and Non-dividing Cells of the Pea Root

The growth of the nucleus and the cell in the pea root was followedthrough the mitotic cycle and subsequently in post-mitotic developmentby comparing cells and nuclei from the meristem, at differentstages of interphase, and cells and nuclei from two regionsof the enlarging zone of the root. Measurements of cell andnuclear volumes were made in sections of fixed roots. Measurementsof nuclear volume, DNA content, and dry mass were made on isolatednuclei. Growth in the mitotic cycle was characterized by a doublingof DNA and nuclear dry mass and a five-fold increase of nuclearvolume. Since cell volume doubled, a differential hydrationof cytoplasm and nucleus is inferred. Post-mitotic growth wascharacterized by a four-fold or greater increase in cell volume,with vacuolation and a continued increase of cytoplasmic constituents,but a cessation of nuclear growth except by uptake of water;the only increase in nuclear dry matter appeared to be in cellsbecoming endopolyploid. The concentration of dry matter in thenucleus fell as the nuclei enlarged in the mitotic cycle andin post-mitotic growth. The relationships between the measuredparameters are examined to see whether they might be indicativeof causal relationships.     

大脑皮质神经干细胞定向分化为神经元过程中Tau蛋白表达的研究

目的探讨Tau蛋白在新生大鼠大脑皮质神经干细胞定向分化为神经元过程中的表达及意义.方法采用细胞培养、免疫细胞化学方法(SABC法)观察及计算机图像分析技术测定Tau蛋白在神经干细胞定向分化为神经元过程中不同时段的表达情况.结果在神经干细胞定向分化为神经元的过程中,Tau蛋白由核周淡染分布渐至在胞体与突起中密集均匀分布,随着突起伸展而不断地延伸,并且Tau蛋白的表达量逐渐增加.结论新生大鼠大脑皮质神经干细胞定向分化为神经元过程中,Tau蛋白的时空表达与神经干细胞定向分化的神经元形态变化有一定的相关性并在此过程中发挥着重要的作用.     

The relationship of DNA synthesis to protein accumulation in the cell nucleus

The relationship between DNA synthesis and protein accumulation in cell nucleus and cytoplasm has been investigated by the use of a combination of ultramicrointerferometric and ultramicrospectrophotometric methods. 5-Fluoro-2'-deoxyuridine (FUdR) inhibited DNA synthesis, resulting in inhibition of cell proliferation in G-1 and early S-phase. However, synthesis and accumulation of protein continued in the presence of FUdR, as indicated by a 54% increase in the average dry mass value per individual cell during 18-hour exposure to FUdR; due primarily to protein accumulation in the cytoplasm, the average cytoplasmic dry mass increased by as much as 85%, while the dry mass of the nucleus increased by only 21%. The dry mass values of individual nuclei were well-correlated to the nuclear DNA content throughout the period of exposure to FUdR. In contrast to the continued accumulation of protein in the cytoplasm during inhibition of DNA synthesis, protein accumulation in the nucleus was inhibited. When cells were released from inhibition of DNA synthesis by the addition of 2'-deoxythymidine, the nuclear DNA content and nuclear dry mass increased in near-synchrony, there being some evidence that DNA synthesis was initiated somewhat prior to initiation of increase in nuclear dry mass. Thus, it appears that DNA synthesis (or an increase in nuclear DNA content) is intimately related to the regulation of protein accumulation in the nucleus.     

Changes in nuclear, nucleolar and cytoplasmic RNA content during growth and differentiation of root parenchyma cells in plant species with different dynamics of DNA endoreplication

Using cytophotometric method, after staining preparations with gallocyanin RNA content was examined in nucleus, nucleolus and cytoplasm of six species of angiospermal plants in successive (1-7 mm) segments of root representing successive zones of differentiation. During the cell cycle, RNA content duplicates in the nucleus, nucleolus and cytoplasm of meristematic cells. On the other hand, during growth and differentiation of parenchyma cells in species with endoreplication the content of nucleolar RNA does not increase in proportion with DNA content. High level of endoreplication is connected with high nucleolar RNA content and low cytoplasmic RNA content. In species without endoreplication at low nucleolar RNA content, a considerable growth of cytoplasmic RNA content takes place.     

Chromosome endoreduplication as a factor of salt adaptation in Sorghum bicolor

Summary. Nuclear DNA amounts were measured by Feulgen cytophotometry in Sorghum bicolor cv. 610 plants early exposed to 150 mM NaCl, a treatment known to induce an increased tolerance to salinity in plants carrying this genotype. In salt-treated plants, the percentages of 8C, 16C, and 32C nuclei in roots in the primary state of growth were 21.9%, 13.3%, and 4.3%, respectively. By contrast, in nonsalinized plants, only 3.5% of the nuclei had an 8C content and no higher DNA contents were observed. The salt treatment induced chromosome endoreduplication during the differentiation of cells in the root cortex, where 41.2% of the cells displayed a DNA content higher than 4C (versus 1.3% in control plants). No enhancement of endopolyploidy was observed in cells of the root vascular cylinder or the leaves of the salt-treated plants. In another S. bicolor genotype (DK 34-Alabama), noncompetent for salt adaptation, the same NaCl treatment did not induce chromosome endoreduplication in root cortex cells. Endopolyploidy may be considered as a part of the adaptive response of S. bicolor competent genotypes to salinity. Correspondence and reprints: Dipartimento di Biologia Cellulare e Ambientale, Università di Perugia, Via A. Pascoli, 06123 Perugia, Italy.     

The distribution of nuclear and cytoplasmic proteins in the blastoderm of the loach, Misgurnus fossilis L

The concentration of dry substance (protein) and the dry weight of nuclei, cytoplasm and cells from different blastoderm regions at the early blastula and midgastrula stages were determined by interferentional microscopy. It was shown that at the early blastula stage the dry weight of cells in the basal layer is higher than that in the outer layer. Although the protein concentration in the basal layer cells appears to be somewhat higher, differences in their dry weight are due primarily to the big volume of cytoplasm of the basal layer cells. By the midgastrula stage, the total (nucleus + cytoplasm) protein concentration increases (by 17% in the basal layer cells and by 9% in the outer layer cells) due to the increase of nuclear protein concentration. At the same time dry weight of these cells markedly decreases due to the decrease of their volumes in the process of cell divisions. At the midgastrula stage the epiblast cells have the highest dry weight due to the highest protein concentration in the cytoplasm and the biggest cell volume. The results obtained are discussed with respect to the data on the pattern of accumulation of newly synthesized protein in nuclei and cytoplasm with special reference to the duration of individual cell cycle phases.     

ENTRY OF BASIC MACROMOLECULES INTO BARLEY ROOTS

The entry of labelled calf-thymus histone, lysozyme, and poly-L-lysine into barley root tips was studied at concentrations which strongly inhibit root elongation. The macromolecules were suitably labelled and at these concentrations it was found, by autoradiography and fluorescence microscopy, that histone and lysozyme readily entered the roots and appeared to bind mainly to cell walls of the epidermis and cortex and to penetrate the cytoplasm occasionally. Except in cap cells, nuclei were rarely penetrated. Poly-L-lysine readily permeated cell walls and invaded cytoplasm and nuclei throughout the root tip. Some cells were damaged by contact with basic macromolecules, as evidenced by a change in appearance of protoplasts under phase contrast and by the inability of these same protoplasts to exclude labelled β-lactoglobulin. Such damage was restricted to cells in contact with the outer solution. Interior to the epidermis, development of many cells was inhibited without visible signs of damage. Evidence supports the conclusion that in the presence of polybasic polymers the integrity of cell membranes is altered, thereby allowing leakage of some cell constituents essential for normal development.     

Interferometric studies of endothelial cells in primary culture

Summary A population of endothelial cells growth from Xenopus laevis tadpole hearts was investigated by microinterferometry. The resulting interferograms were evaluated by an automatic image analyzer. The mean values of dry mass were 778±340 pg (10^–15 g) for whole cells, 648±309 pg for cytoplasm, 116±45 pg for nuclei, and 19±10 pg for nucleoli. Two subpopulations of cells were identified, an actively growing one and a less active one. The density (dry mass per m²) of the nuclei and nucleoli of less active cells was greater than that of the nuclei and nucleoli of actively growing cells. In addition, the inactive cells were always large and possessed a considerable amount of cytoplasm. The entrance of cells into S-phase could not be detected by microinterferometry; and no differences were apparent between cells possessing one nucleolus and those containing two nucleoli. The values obtained in these amphibian cells were compared with those derived from mammalian cells.     

Nuclear and cytoplasmic changes during early stages of cell differentiation in roots of the water-fern,Azolla pinnata

Summary Selected nuclear and cytoplasmic changes associated with early differentiation of four cell-types—dermatogen, inner and outer cortex, and endodermis—have been analysed using montages of electron micrographs of median longitudinal sections of young roots ofAzolla pinnata. The area fraction of nucleoplasm occupied by chromocentres (CAF) is smaller in the apical cell than in the nuclei of its most recently formed daughter cells. The CAF also differs between the four cell-types: dermatogen nuclei have a lesser mean CAF and smaller chromocentres than nuclei of the endodermis; cortical cell nuclei have intermediate values. These differences may reflect changes in nuclear activity during cell differentiation. The area fraction occupied by the vacuome (VAF) differs between the apical cell and its daughters: the apical cell seems to retain most of the vacuome at division, while the daughter cells receive less vacuolate cytoplasm. Of the four cell-types analysed, the cortical cells develop a large VAF the quickest; the dermatogen is slower to become vacuolate. Cells in the dermatogen and outer cortex derive from common mother cells, as do cells in the endodermis and inner cortex, and even the most recently-formed cells in the files of inner and outer cortex are more vacuolated than their sister cells in the other two celltypes. The onset of vacuolation may be triggered by an inductive influence emanating from older vacuolated cells in the same file. The rate of vacuolation in each of the cell-types examined may also be negatively correlated to the intensity of synthesis of protein used to construct cytoplasmic materials.     

用胶体金免疫电镜技术研究水分胁迫对蚕豆根中ABA分布与含量的影响

用胶体金免疫电镜技术对蚕豆 ( Vicia faba L.)根中 ABA的定位结果表明 ,在原分生组织金颗粒主要分布在细胞核。在基本分生组织或伸长区前部皮层细胞的质膜附近有较多的金颗粒标记 ,维管柱特别是维管组织的质外体中发现有大量金颗粒标记。伸长区中部及根毛区细胞也有较多的金颗粒标记。水分胁迫可导致初生分生组织或伸长区前部细胞金颗粒标记密度大增 ,伸长区中部及根毛区细胞的金颗粒密度也明显增加。对根中 ABA在超微结构水平上的分布及其与运输的关系进行了讨论 ,为 ABA有可能作为由根向地上部分传递的逆境信息提供了进一步的证据。     

Activities of DNA polymerases and RNA polymerases detected in situ in growing and differentiating cells of root cortex.

Activities of DNA polymerases and RNA polymerases were studied by autoradiographic methods in growing and differentiating root cortex cells of Zea mays - a species in which endomitosis occurs - and Tulipa kaufmanniana - in which this process does not occur. In Tulipa kaufmanniana, the highest activity of DNA polymerase appears in the nuclei of meristematic zone during the S phase of the cell cycle. In Zea mays, endomitotic replication of DNA occurs in all growth and differentiation zones and the activity of DNA polymerase in the nuclei is similar to that in the meristematic zone. In both species, nuclear RNA synthesis, measured with 3H uridine incorporation, is highest in the meristematic zone and declines steadily with development. Activity of nuclear RNA polymerase is present in all developmental zones in both species and is similar to that in the meristematic zone. 3H uridine incorporation into nucleoli decreases markedly in both species, whereas the activity of nucleolar RNA polymerase remains at a high level in all root segments in Zea mays and decreases slightly in Tulipa kaufmanniana. It is argued that the differences between the incorporation of 3H uridine and that or 3H UMP may be caused by a reduction of the pool of endogenous ribonucleoside triphosphates. Marked activities of DNA polymerase and RNA polymerase in cytoplasm are possibly related to the growth and division of plastids and mitochondria.     

An Immunogold Microscopic Study on the Effects of Water Stress on ABA Localization and Contents in Roots of Vicia faba

ABA localization in roots of Vicia faba L. was studied using immunogold microscopy. In cells of promeristem gold particles were mainly localized in the nuclei. In cells of ground meristem and cortex of the front part of elongation zone, some gold particles were found in cytoplasm near. the plasmalemma. Substantial amounts of gold particles were observed in cells of vascular cylinder especially in apoplast of vascular tissue. Cells of middle elongation zone and root hair zone were also labelled by many gold particles. In cells of the primary meristem and the front part of elongation zone, water stress could lead to acute increase of the gold particle density, and also in the cells of the elongation and root hair zone. The distribution of ABA in subcellular level and its relationship with transportation were discussed in the text. and the results provided evidence for ABA as a root-to-shoot transporting stress signal.     

Cellular distribution of lens epithelium-derived growth factor (LEDGF) in the rat eye: loss of LEDGF from nuclei of differentiating cells

Lens epithelium-derived growth factor (LEDGF) enhances the survival and growth of cells. To understand LEDGF's spatial localization and its putative function(s) during proliferation and differentiation, we localized LEDGF during terminal differentiation in whole rat lenses, lens epithelial cell (LEC) explants stimulated with FGF-2, and insulin, iris, human LECs with lentoids. In addition, intracellular localization of LEDGF was performed in other ocular tissues: ciliary body, retina, and cornea. We found the immunopositivity of nuclear LEDGF decreased in LECs of the equatorial region. In contrast, immunopositivity of LEDGF was detected in the cytoplasm of LECs and superficial fiber cells. After treating LEC explants with FGF-2 and insulin, which are known to be differentiating factors for LECs, the nuclei of these cells showed no LEDGF immunopositivity, but explants did express p57(kip2), a differentiation marker protein. Also, immunopositive LEDGF was not detected in the nuclei of differentiated cells, lentoid body, and corneal epithelial cells. This demonstrated that the loss of LEDGF from the nucleus may be associated with the process of terminal differentiation that might be in some way common with the biochemical mechanisms of apoptosis. The spatial and temporal distribution of LEDGF in the present study also provides a vision for further investigation as to how this protein is involved in cell fate determination.     

The Relative Concentration of Solids in the Nucleolus, Nucleus, and Cytoplasm of the Developing Nerve Cell of the Chick

Growing and differentiating nerve cells of the fifth cranial ganglion of the chick embryo were studied by several means. During the period of 70 hours to 11 days of incubation (Hamburger-Hamilton stages 19 to 37) average cell mass increased more than 4.5 times while cells changed from relatively undifferentiated neuroblasts to morphologically characteristic nerve cells with long processes. By making simplifying assumptions about thickness of nucleus and nucleolus, relative to cytoplasmic thickness, it was possible to calculate solute concentration of nucleus and nucleolus relative to that of the cytoplasm from measurements of optical retardations through living cells. Differences in relative solute concentration were observed in nucleolus, cytoplasm, and nucleoplasm in the approximate ratio 1.2:1.0:0.8, respectively. The ratio remained essentially constant during the growth period examined despite the fact that the cell components grow at markedly different rates. This suggests that solid concentrations are physical characteristics of nucleus, nucleolus, and cytoplasm which are maintained even during rapid growth and differentiation. By cytochemical means it was demonstrated that mass increase in the nucleus is not associated with increase in deoxyribonucleic acid. Both ribonucleic acid and protein are in greater concentration in nucleolus and cytoplasm than in the nucleoplasm. Electron microscopy shows interruptions in the nuclear envelope as well as an approximately even distribution of electron density in nucleus and cytoplasm. It is pointed out that consistent differences in solid concentration can exist on either side of the nuclear envelope even though it contains "pores." Implications of these data are discussed.     

Changes in nuclear and nucleolar protein content during the growth and differentiation of root parenchyma cells in plant species with different DNA-endoreplication dynamics

Using cytophotometric procedures, we measured the nuclear and nucleolar protein content of successive zones of growth and differentiation in consecutive (1-7 mm) root segments obtained from eight species of the Angiospermae after staining the preparations with Feulgen-Naphthol Yellow S (F-NYS). In meristematic cells the nuclear and nucleolar protein content was found to double during the cell cycle. In species in which differentiation occurs at the same time as nuclear DNA endoreplication, i.e. Vicia faba subsp. minor, V. faba subsp. major, Pisum sativum, Hordeum vulgare and Amaryllis belladonna, the pool of nuclear proteins observed during the G2 phase of the cell cycle was seen in the differentiated zone in nuclei containing 8C DNA. Species in which differentiation is not accompanied by the process of nuclear DNA endoreplication, i.e. Levisticum officinale, Tulipa kaufmanniana and Haemanthus katharinae, exhibited the highest nuclear proteins content during the G2 phase of the cell cycle; comparably high values were not found in the differentiated zone. A decrease in nucleolar protein content was observed during the process of differentiation, this tendency being more evident in the studied species that do not exhibit endoreplication.     

The nuclear dry weight of liver cells from patients with virus hepatitis and from controls.

Interferometric investigations were performed at liver cell nuclei isolated in 70 per cent glycerol. In 11 patients with virus hepatitis and seven patients without liver disease the nuclear dry weight of liver cells obtained by liver biopsy was determined by interferometry. The average nuclear dry weight of diploid liver cells from controls was 39.9 pg while an average value of 45.4 pg was found for patients with hepatitis. The corresponding nuclear volumes were 241 and 274mu3 respectively. The dry mass and volume of tetraploid nuclei was twice as big as that of diploid nuclei in both materials. The nuclear water content was neither significantly different between diploid and tetraploid nuclei nor significantly different between nuclei from controls and patients with hepatitis.     

Incorporation of regulatory peptide hormones by individual cells of the adrenal cortex: Prolactin-adrenocorticotrophin differences

Recent refinements in methodology now permit the study of endogenous peptide hormones in their individual target cells. The investigations reported here deal with the question of whether endogenous ACTH can be detected in its target cells in the highly active adrenal gland of the normally lactating rat. This question was examined with immunohistochemistry. ACTH was found in both cytoplasm and nuclei of adrenal glomerulosa cells. In cells of the fasciculata and reticularis layers of the adrenal cortex, it did not appear inside nuclei but was present in the cytoplasm and on the nuclear envelope. The distribution of ACTH was compared with and found to be different from that of PRL. PRL, confirming previous findings, was not detectable at all in glomerulosa cells and, in cells of the inner cortical zones, was present in both cytoplasm and nuclei. In neither case was hormone found in the adrenal medulla. The apparent feasibility of studying peptide regulators such as ACTH and PRL in their individual target cells may be a focal point for an acceleration of our understanding of how these peptides work.     

Compartmentation and Fluxes of Sugars in Roots of Phaseolus Vulgaris Under Phosphate Deficiency

The influence of phosphate deficiency on the sugar accumulation and sugar partitioning in the root cells of bean (Phaseolus vulgaris L.) was studied. Bean plants were cultured 17 - 19 d on a phosphate-sufficient and phosphate-deficient nutrient medium. Phosphate deficit in the growth medium resulted in increased sugar concentration for about 30 % in the apoplastic and cytoplasmic compartments as well as in the vacuoles of root cells. However, the distribution of sugars between apoplast and cytoplasm compartment and vacuole was not affected by decreased phosphate concentration. About 20 % of sugars were found in the apoplast and cytoplasm, about 80 % in the vacuole. Low phosphate concentration enhanced influx of exogenous ¹⁴C-sucrose into meristematic and elongation zones of root. The ¹⁴C-labelled sugar content in the root tips increased for about 60 % as compared to control plants. Phosphate deficiency increased also ¹⁴C-glucose uptake and content in the root tips. However, the amount of ¹⁴CO₂ liberated during respiration of P-deficient roots (after feeding with uniformly labelled ¹⁴C-glucose) was lower than ¹⁴CO₂ respired by control plants, thus a large part of accumulated sugars seems to be metabolically inactive. This revised version was published online in July 2006 with corrections to the Cover Date.