Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “CYTOPLASM”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 919 records · Page 51Linked to original sources
PubMed1994

Expression of cytoplasmic and nuclear vasopressin RNA following castration and testosterone replacement: evidence for transcriptional regulation.

In order to determine the mechanism by which testosterone (T) regulates the expression of vasopressin (VP) in the bed nucleus of the stria terminalis (BNST) and medial amygdala (MA), in situ hybridization's were performed to measure the expression of cytoplasmic VP mRNA and nuclear VP RNA in Long-Evans rats that were castrated or T replaced for various lengths of time. One hour after castration, plasma T levels were significantly reduced and remained at this low level until the T was replaced in animals. Three hours after T replacement, plasma T levels were significantly elevated compared to those of control animals and returned to control levels within 9 h of replacement. Modulation of nuclear VP RNA expression by castration and T replacement was rapid. Six hours after castration, levels of nuclear VP RNA in the BNST and MA were reduced below limits of detection. Three hours after T replacement nuclear VP RNA expression was elevated in BNST and MA, achieving levels comparable to controls after 9 h of T replacement. In the BNST, the number of cells positively labeled by the nuclear VP RNA probe closely paralleled the changes observed in plasma T; in that 24 h after T replacement there was a significant increase in the number of cells positively labeled for nuclear VP RNA, with control levels returning 3 days later. Cytoplasmic VP mRNA expression in the BNST and MA, on the other hand, was slower to respond to both castration and T replacement than nuclear VP RNA. A significant change in the number of cells expressing cytoplasmic VP mRNA was observed approximately 3 days after castration in the BNST and 7 days after castration in the MA. The amount of labeling per cell of cytoplasmic VP mRNA responded more rapidly to the reduction of circulating T than the number of positively labeled cells. A significant decrease in grains/cell was observed 24 h after castration in the BNST and 3 days after castration in the MA. The number of cells expressing VP mRNA in both the BNST and MA required 3 days of T replacement to return to control levels; while the amount of labeling per cell required 7 days of replacement. Therefore, these data suggest that T's ability to regulate the expression of the VP mRNA occurs at least in part at the transcriptional level, since the expression of VP primary transcript was altered prior to changes in the hybridization signal for the cytoplasmic VP mRNA.

Amygdala↗
PubMed1995

An extranuclear expression system for analysis of cytoplasmic promoters of yeast linear killer plasmids.

Based on the cytoplasmically localized killer plasmids pGKL1 and pGKL2 of Kluyveromyces lactis two new linear hybrid plasmids were constructed which consist of pGKL1, into which in addition to the previously developed cytoplasmically expressible LEU2* selectable marker a glucose dehydrogenase-encoding bacterial gene (gdh A) has been integrated. One of the hybrid plasmids carries the bacterial gene preceded by an arbitrarily placed cytoplasmic promoter (upstream conserved sequence) in front of the coding region (pRKL121). The other plasmid was constructed in such a way that the ATG start codon of the gdh A gene was fused in frame to the ATG start codon of the killer plasmid's open reading frame 5 (pRKL122). The structures of both linear hybrid plasmids were confirmed by restriction analysis, Southern hybridization, and sequencing of the junction sites. Yeast strains carrying either of the plasmids expressed the glucose dehydrogenase gene; however, expression of the in phase fused gene was 40-fold higher compared to the arbitrarily placed cytoplasmic promoter. In general, an in phase fusion was not required for expression, but efficiency is dramatically enhanced when the 5' noncoding sequences in front of the heterologous genes are the same as those found on the native killer plasmids. The developed system can serve as a reporter for determining the efficiency of the different cytoplasmic promoters present on both linear plasmids. Hybrid plasmids were stably maintained without selective pressure in K. lactis and they were transferred and expressed also in Saccharomyces cerevisiae.

Bacillus megaterium↗
PubMed2001

Production of correctly folded Fab antibody fragment in the cytoplasm of Escherichia coli trxB gor mutants via the coexpression of molecular chaperones.

Disulfide bonds are normally formed after a polypeptide has been exported from the reducing environment of the cytoplasm into a more oxidizing compartment, such as the bacterial periplasm. Recently, we showed that in Escherichia coli trxB gor mutants, in which the reduction of thioredoxin and glutathione is impaired, the redox potential of the cytoplasm becomes comparable to that of the mammalian endoplasmic reticulum, thus allowing the formation of disulfide bonds in certain complex proteins (P. H. Bessette et al., 1999, Proc. Natl. Acad. Sci. USA 96, 13703-13708]. Here, we investigate the expression of a Fab antibody fragment in the bacterial cytoplasm. The effect of coexpressing cytoplasmic chaperones (GroEL/ES, trigger factor, DnaK/J), as well as signal sequenceless versions of periplasmic chaperones (DsbC and Skp), was examined. Skp coexpression was shown to have the most significant effect (five- to sixfold increase) on the yield of correctly folded Fab. A maximum yield of 0.8 mg Fab/L/OD(600) Fab was obtained, indicating that cytoplasmic expression may be a viable alternative for the preparative production of antibody fragments.

Amino Acid Sequence↗
PubMed1994

The HSV-1 2-kb latency-associated transcript is found in the cytoplasm comigrating with ribosomal subunits during productive infection.

We have examined the nuclear and cytoplasmic distribution of the latency-associated transcripts (LATs) of HSV-1. During latency these transcripts accumulate in the nuclei of neurons in the peripheral and central nervous system of infected animals. However, our Northern blot analyses demonstrate that the 2-kb LAT is found in the cytoplasm of HSV-1-infected CV-1 cells, and brainstems of HSV-1 productively infected mice. Like the nuclear LAT from latently infected tissue, most of the cytoplasmic 2-kb LAT from lytically infected CV-1 cells is unpolyadenylated. In order to determine if cytoplasmic LAT could be translated, we compared its distribution with that of glycoprotein C mRNA in polysome profiles from HSV-1-infected tissue culture cells. Specific association of RNAs to polysomes was verified by disruption of polysomes with EDTA or puromycin. Analyses of numerous experiments indicate that most of the cytoplasmic 2-kb LAT migrates at the position of ribosomal subunits in polysome profiles. Thus, the 2-kb LAT may not be efficiently translated during productive infection. This suggests that if the 2-kb LAT is indeed translated, its translation may be tightly regulated during HSV-1 infection, possibly in a cell type- or cell cycle-specific manner. Another possibility is that the 2-kb LAT is not a translated RNA but may have another function, possibly related to translation as indicated by its apparent association to ribosomal complexes.

Animals↗
PubMed1979

Ultrastructure and cytochemistry of the yolk syncytial layer in the alevin of trout (Salmo fario trutta L. and Salmo gairdneri R.) after hatching. II. The cytoplasmic zone.

In contrast to the vitellolysis zone which is involved in the degradation of the yolk, the cytoplasmic zone of the yolk syncytial layer, composed of many cytoplasmic organelles, is implicated in a secretory process. The granular endoplasmic reticulum of the latter is extremely well developed and organized into trabeculae. The yolk nuclei show a RNA positive reaction. The Golgi apparatus is implicated in the elaboration of very low density lipoproteins (VLDL) and of acid phosphatase. The numerous mitochondria seem to suggest an important energy metabolism in the solubilisation area. The appearance of certain special structures (crystalline bodies, pseudovesicular structures, lipochondria) as well as their relation with the organelles of the cytoplasmic zone, re-inforces the impression of a layer with a secretory nature and suggests its participation in the remodelling of the yolk products produced in the vitellolysis zone. The results of the investigations concerning the acid phosphatase activity suggest that this enzyme plays a role on the one hand in the degradation of certain platelets which penetrate into the cytoplasmic zone, and on the other hand in the regulation of VLDL, in the lysis of secretory products elaborated by the cytoplasmic zone. The possible presence of microperoxisomes is discussed as well as the positive detection of glucose-6-phosphate dehydrogenase.

Acid Phosphatase↗
PubMed1992

Activity of cardiac L-type Ca2+ channels is sensitive to cytoplasmic calcium.

Ca(2+)-induced inactivation of L-type Ca2+ channels is proposed as an important negative feedback mechanism regulating Ca2+ entry. Here, for the first time, evidence for modification of heart L-type Ca2+ channel activity by cytoplasmic calcium is provided from excised inside-out membrane patches. Ba2+ currents through cardiac L-type Ca2+ channels exhibited only modest inactivation in the absence of cytoplasmic Ca2+. Elevation of cytoplasmic Ca2+ to micromolar concentrations strikingly affected L-type Ca2+ channel activity as evaluated from ensemble average Ba2+ currents. Inactivation was markedly increased concomitant with a reduction of peak inward current, which was almost completely eliminated at about 15 microM cytoplasmic Ca2+ concentration. Half maximal suppression of Ba2+ currents was observed at 2.3 microM Ca2+. The observed modifications of L-type Ca2+ channel activity show that cytoplasmic Ca2+ induces channel closure. Below 4 microM Ca2+, channels can be reversibly reactivated during repetitive depolarizations, while at high Ca2+ concentrations (approximately 15 microM) most Ca2+ channels reside in a closed state. This may allow for a delicate regulation of Ca2+ entry, and consequently of heart contraction.

Animals↗
PubMed1986

Heterogeneity of circular mitochondrial DNA molecules from sugar beet with normal and male sterile cytoplasms.

Mitochondrial (mt) DNAs from normal (N) and male sterile (S) cytoplasms of sugar beet have been isolated and investigated by electron microscopy. The results showed that mtDNA was composed of a heterogeneous population of circular molecules. Their contour lengths varied from 0.28 to 51 micron, but unlike in the case of maize, a large difference was not observed in the distribution of molecular classes greater than 1.0 micron between N and S cytoplasms of sugar beet. On the other hand, N and S cytoplasms were shown to contain their own characteristic combinations of small circular mtDNA species with lengths between 0.28 micron and 0.6 micron. Mitochondrial DNAs from various sources of male-sterile cytoplasms were analyzed by agarose gel electrophoresis to determine the extent of cytoplasmic variation. Additional low molecular weight DNA bands appeared in all male-sterile lines examined, and as a result, three distinctive banding patterns were recognized. These data are in general agreement with those based upon restriction endonuclease digestion of mt and chloroplast DNAs and the genetic analysis of fertility restoration in test crosses.

Cytoplasm↗
PubMed1977

Blue light-dependent regulation of cytoplasmic ribosomal RNA synthesis in Chlorella.

Effect of blue and red light on ribosomal RNA synthesis in autotrophic synchronous cultures of Chlorella pyrenoidosa (strain 211-8b) is studied by pulse labeling experiments with tritiated guanosine. Nucleic acids were separated by electrophoresis on polyacrylamide gels. Compared with darkness and red light (679 nm), blue light (457 nm) of equal quantum flux (0.5-5x10(-10) Einstein cm-2 s-1) stimulates incorporation into ribosomal RNA. This blue light effect is observed in the cytoplasmic ribosomal RNA after 5 min of illumination, whereas the stimulation of chloroplast ribosomal RNA synthesis by blue light appears later. Maturation of chloroplast ribosomal RNA is slower than that of cytoplasmic ribosomal RNA. The blue light effect on the cytoplasmic ribosomal RNA formation does not require chloroplast RNA or protein synthesis as shown by inhibitor studies with rifampicin or lincomycin. The blocking of cytoplasmic protein synthesis by cycloheximide inhibits the blue light effect on ribosomal RNA formation. It is concluded that the cytoplasmic ribosomal RNA transcription is controlled by a blue light sensitive system.

Chlorella↗
PubMed1980

Suppressive action of cytoplasmic and metabolite extracts of Candida albicans on the immune response in guinea pigs.

This study investigated the possibility that Candida albicans components exert a suppressive effect on the immune response of guinea pigs (GP), similar to that of live C. albicans organisms as was previously shown. Hartley GP were inoculated with C. albicans crude cytoplasmic or metabolite (culture filtrate) extracts (containing most of the organism's cell contents or its metabolite and degradation products, respectively). Their immunological responses towards sheep red blood cells (SRBC) were compared with those of GP inoculated with SRBC alone or with SRBC together with live C. albicans organisms. The immunological responses were measured by: 1) rosette formation (RF) of SRBC with peritoneal macrophages, 2) haemolytic plaque formation (PFC) with lymph node-cells, 3) haemagglutination and 4) haemolysis tests. According to the RF tests, inoculation of GP with either cytoplasmic or metabolite extracts resulted in decreased RF as compared to GP inoculated with SRBC only; the decrease was correlated with the protein concentration of the extracts. Inoculation with metabolite extract led to a more diminished RF than with cytoplasmic extract, but less than with live C. albicans organisms. Inoculation of cytoplasmic extracts did not affect the haemagglutinin and haemolysin titers, while that of metabolite extracts resulted in a slight decrease of these titers. The assays for PFC were not conclusive enough to point to a suppressive effect of C. albicans extracts. In summary, it appears that both the cytoplasmic and metabolite extracts of C. albicans exert a partial suppressive effect on the immune response in GP, as judged primarily on the basis of the RF results.

Animals↗
PubMed1982

Expression of common cytoplasmic antigens by epithelial cells of thymus and epidermis.

The expression of common cytoplasmic antigens by thymic epithelial cells and keratinocytes was analyzed by immunolabelling on cryostat sections of human and animal thymus and epidermis. Experimental sera to human epidermal keratin subunits of molecular weight (MW) 67 K, 63 K and 55 K were used, as well as human sera with antibodies to epidermal cytoplasmic antigens (KCA), reacting either with U-Cyt antigens expressed by cells from the upper compartment of the epidermis or with BLC antigens in cells from the basal layer. It was demonstrated that keratin subunits of MW 67 K and 63 K were detected only in the upper layers of human epidermis, whereas 55 K was present in all epidermal layers. On rabbit lip a labelling was obtained throughout the epidermis by the three immune sera. On thymus sections a cytoplasmic staining of epithelial cells was observed with the three immune sera, suggesting an antigenic similarity between cytokeratins of thymus epithelial cells and keratins of the upper layers of human epidermis. Two human sera with U-Cyt antibodies and one with BCL antibodies labelled human thymic epithelial cells but were negative with mice thymus. Ten other sera with KCA were negative on both thymus. These results show that the expression of keratins and cytoplasmic antigens, defined by human KCA, differed in immunofluorescence on normal human epidermis, rabbit lip and thymic epithelial cells. These suggests a heterogeneity among human keratinocyte cytoplasmic antigens defined by KCA and the presence in man of common antigenic constituents from keratins in epidermal cells and thymic epithelial cells.

Animals↗
PubMed1995

Detection of gravity-induced polarity of cytoplasmic streaming in Chara.

Gravity induces a polarity of cytoplasmic streaming in vertically-oriented internodal cells of characean algae. The motive force that powers cytoplasmic streaming is generated at the ectoplasmic/endoplasmic interface. The velocity of streaming, which is about 100 micrometers/s at this interface, decreases with distance from the interface on either side of the cell to 0 micrometers/s near the middle. Therefore, when discussing streaming velocity it is necessary to specify the tangential plane through the cell in which streaming is being measured. This is easily done with a moderate resolution light microscope (which has a lateral resolution of 0.6 micrometers and a depth of field of 1.4 micrometers), but is obscured when using any low resolution technique, such as low magnification light microscopy or laser Doppler spectroscopy. In addition, the effect of gravity on the polarity of cytoplasmic streaming declines with increasing physiological age of isolated cells. Using a classical mechanical analysis, we show that the effect of gravity on the polarity of cytoplasmic streaming cannot result from the effect of gravity acting directly on individual cytoplasmic particles. We suggest that gravity may best be perceived by the entire cell at the plasma membrane-extracellular matrix junction.

Cell Polarity↗
PubMed2001

Cytoplasmic acidification with butyric acid does not alter the ionic conductivity of plasmodesmata.

The effect of lowering cytoplasmic pH on the ionic conductivity of higher-plant plasmodesmata was investigated with corn (Zea mays L. cv. Black Mexican Sweet) suspension culture cells. Exposure to butyric acid decreased the cytoplasmic pH by 0.8 units. Intercellular communication was monitored by electrophysiological techniques that allowed the measurement of membrane resistances of sister cells and the electrical resistance of the plasmodesmata connecting them. The decrease in cytoplasmic pH did not affect the resistance of plasmodesmata, despite the fact that the butyric acid treatment more than doubled the concentration of cytoplasmic calcium. This is discussed in light of previous findings that increases in cytoplasmic calcium increase the electrical resistance of plasmodesmata.

Butyric Acid↗
PubMed1990

Gravity-dependent polarity of cytoplasmic streaming in Nitellopsis.

The internodal cells of the characean alga Nitellopsis obtusa were chosen to investigate the effect of gravity on cytoplasmic streaming. Horizontal cells exhibit streaming with equal velocities in both directions, whereas in vertically oriented cells, the downward-streaming cytoplasm flows ca. 10% faster than the upward-streaming cytoplasm. These results are independent of the orientation of the morphological top and bottom of the cell. We define the ratio of the velocity of the downward- to the upward-streaming cytoplasm as the polar ratio (PR). The normal polarity of a cell can be reversed (PR < 1) by treatment with neutral red (NR). The NR effect may be the result of membrane hyperpolarization, caused by the opening of K+ channels. The K+ channel blocker TEA Cl- inhibits the NR effect. External Ca2+ is required for normal graviresponsiveness. The [Ca2+] of the medium determines the polarity of cytoplasmic streaming. Less than 1 micromole Ca2+ resulted in a PR < 1 while greater than 1 micromole Ca2+ resulted in the normal gravity response. The voltage-dependent Ca(2+)-channel blocker, nifedipine, inhibited the gravity response in a reversible manner, while treatment with LaCl3 resulted in a PR < 1, indicating the presence of two types of Ca2+ channels. A new model for graviperception is presented in which the whole cell acts as the gravity sensor, and the plasma membrane acts as the gravireceptor. This is supported by ligation and UV irradiation experiments which indicate that the membranes at both ends of the cell are required for graviperception. The density of the external medium also affects the PR of Nitellopsis. Calculations are presented that indicate that the weight of the protoplasm may provide enough potential energy to open ion channels.

Calcium↗
PubMed1991

Cytoplasmic calcium levels in protoplasts from the cap and elongation zone of maize roots.

Calcium has been implicated as a key component in the signal transduction process of root gravitropism. We measured cytoplasmic free calcium in protoplasts isolated from the elongation zone and cap of primary roots of light-grown, vertically oriented seedlings of Zea mays L. Protoplasts were loaded with the penta-potassium salts of fura-2 and indo-1 by incubation in acidic solutions of these calcium indicators. Loading increased with decreasing pH but the pH dependence was stronger for indo-1 than for fura-2. In the case of fura-2, loading was enhanced only at the lowest pH (4.5) tested. Dyes loaded in this manner were distributed predominantly in the cytoplasm as indicated by fluorescence patterns. As an alternative method of loading, protoplasts were incubated with the acetoxymethylesters of fura-2 and indo-1. Protoplasts loaded by this method exhibited fluorescence both in the cytoplasm and in association with various organelles. Cytoplasmic calcium levels measured using spectrofluorometry, were found to be 160 +/- 40 nM and 257 +/- 27 nM, respectively, in populations of protoplasts from the root cap and elongation zone. Cytoplasmic free calcium did not increase upon addition of calcium to the incubation medium, indicating that the passive permeability to calcium was low.

Calcium↗
PubMed1987

Scanning electron microscopic studies of the myelinated nerve fibres of the mouse sciatic nerve with special reference to the Schwann cell cytoplasmic network external to the myelin sheath.

The three-dimensional morphology of the surface of myelinated nerve fibres in the mouse sciatic nerve was studied by scanning electron microscopy after combined potassium hydroxide treatment and collagenase digestion (to remove the surrounding collagen fibrils and basal laminae from nerve fibres) as well as by transmission electron microscopy. The myelinated nerve fibre appeared as a long cylinder with sporadic annular constrictions corresponding to the nodes of Ranvier. Slight swellings of the surface due to Schwann cell nuclei were usually found at the middle of each internode. The surface of the nerve fibre clearly exhibited a network of bulges, which consisted of longitudinal bands extending from the nuclear swelling to the nodes of Ranvier through the internode, and transverse trabeculae bridging between these longitudinal bands. These bulges on the surface of nerve fibres were the site of the retained Schwann cell cytoplasm external to the myelin lamellae. These cytoplasmic networks on myelinated fibres presumably corresponded to the networks described by Cajal following silver impregnation. In addition, other thin elevations and focal round swellings were also found associated with these longitudinal bands and transverse trabeculae. These networks of Schwann cell cytoplasm are considered to be cytoplasmic channels for nutrition. The two apposing paranodal bulbs of nodes of Ranvier were often asymmetrical in their structure. The networks of the paranodal region were more complicated than those in the internode. The networks of Schwann cell cytoplasm converged into a continuous circumferential collar toward the node, which in turn gave rise to finger-like projections into the nodal gap.

Animals↗
PubMed1977

Comparative studies on the '5'-cap' and in vitro translational activity of cytoplasmic and nuclear poly A(+) RNA1.

The translational activities of cytoplasmic poly A(+)RNA of normal rat liver and Novikoff hepatoma cells in the wheat germ cell free system were found to be approximately 15-20 times greater than tose of the corresponding nuclear poly A(+) RNA. The translationsl activities were 85 and 62 pmoles 3H-leucine incorporated/micron g cytoglasmic poly A(+) RNA for the liver and tumor respectively and 3-4 pmoles 3H-leucine incoporated/micron g nuclear poly A(+)RNA. Inasmuch as intergity of the '5'-cap' of mRNA is essential for its translational activity, quantitative comparisons were made of its content in these RNA fractions. Of the total 32P incorporated into the tumor cytoplasmic poly A(+) RNA, 0.41% was in the '5'-cap'; in nuclear poly A(+) RNA, the '5'-cap' contained 0.11%. After periodate oxidation and labeling with KB3H4, m7 guanosine, the 5'-terminal nucleoside in both liver and Novikoff hepatoma nuclear poly A(+) RNA contained approximately 20% as much isotope as in the cytoplasmic poly A(+) RNA. These results suggest the lower translational activity of nuclear poly A(+) RNA is partly related to its lower content of the '5'-cap'. Molecular selection of poly A(+) RNA for transport out of the nucleus or further cytoplasmic processing may account for the higher percentage of the '5-cap' and the greater translational activity of the cytoplasmic poly A(+) RNA. During these studies, it was also found that the m7 guanosine of the '5'-cap' was not removed during translation of the mRNA in the wheat germ system; this result suggests that the '5'-cap' may associate with allosteric binding sites of initiation factor(s).

Animals↗
PubMed1977

Cytoduction as a new tool in studying the cytoplasmic heredity in yeast.

When crossing the haploid cells of genetically marked yeast strains we observed the appearance of both normal diploid zygotes and haploid nuclear cytoplasmic hybrids. The latter had the nuclear markers of one and the cytoplasmic marker (rho+) of the other parent. The autonomous cytoplasmic factor transfer was termed as cytoduction. Cytoduction is supposed to be the abortive form of yeast cell mating. Only about 1% of cytoductants in usually observed. Cytoduction can be used as a simple test on cytoplasmic determination of some characters. We observed the transfer into cytoductant cells of not only rho+ marker but of resistance factors to antibiotics (erythromycin, neomycin) and killer factor as well. Cytoduction can be applied towards constructing strains having the identical nucleus genotype with mitochondria and other cytoplasmic factors of different origin. In crossing strains with doubly marked mitochondria recombination of mitochondrial markers in cytoductant haploid cells was observed, the pattern of which was similar to that of mitochondrial recombination in normal zygotes.

Crosses, Genetic↗
PubMed1996

The importance of cytoplasm in early embryonic development.

PURPOSE: A mouse model was used to evaluate the consequences of partial cytoplasm removal, which mimics the consequence of human early embryo fragmentation, at the one-cell stage on embryonic development to the morula and blastocyst stages. RESULTS: Opening the zona pellucida at the zygote stage has no detrimental effect on the number of differentiated or total cells at morula/blastocyst stage. However, reduced cytoplasm significantly affects the number of inner cell mass cells, 21.7 (10% cytoplasm removed) vs 27.6 (control) and 15.1 (20% cytoplasm removed) vs 24.7 (control). CONCLUSIONS: The contribution of cytoplasm of maternal origin is critical to early embryonic development.

Animals↗