scholarly journals Changes in the nuclear polyamine content of chick erythrocytes during embryonic development

1979 ◽  
Vol 184 (3) ◽  
pp. 607-612 ◽  
Author(s):  
M H Goyns
Keyword(s):  
Embryonic Development ◽  
Metabolic Activity ◽  
Embryonic Chick ◽  
Polyamine Content ◽  
In Cells

The polyamine content of the circulating erythrocyte population in the embryonic chick was studied during its development. Total cellular polyamine content fell dramatically between 5 and 7 days of development, paralleling the decrease in metabolic activity exhibited by these cells. Nuclei were isolated from the erythrocytes by a non-aqueous technique, which not only eliminated the polyamine loss that occurred with aqueous isolation, but also prevented redistribution of the polyamines from the cytoplasm. Nuclear spermidine and spermine contents decreased markedly between 5 and 6 days of development from 31 to 10 pmol/microgram of DNA and from 33 to 18 pmol/microgram of DNA respectively. Thereafter the spermine content remained constant, but the spermidine content continued to decline. Good correlations between spermidine and RNA contents were observed in both cells and nuclei, and similarly between spermine and RNA contents in cells, but no such correlation was observed between spermine and RNA in nuclei.

scholarly journals The arginine metabolite agmatine protects mitochondrial function and confers resistance to cellular apoptosis

2009 ◽  
Vol 296 (6) ◽  
pp. C1411-C1419 ◽  
Author(s):  
Mary Ann Arndt ◽  
Valentina Battaglia ◽  
Eva Parisi ◽  
Mark J. Lortie ◽  
Masato Isome ◽  
...  
Keyword(s):  
Free Radical ◽  
Chromatin Condensation ◽  
Free Radical Scavenger ◽  
Radical Scavenger ◽  
3T3 Cells ◽  
Polyamine Content ◽  
Cellular Apoptosis ◽  
Models Of Injury ◽  
Nih 3T3 ◽  
In Cells

Agmatine, an endogenous metabolite of arginine, selectively suppresses growth in cells with high proliferative kinetics, such as transformed cells, through depletion of intracellular polyamine levels. In the present study, we depleted intracellular polyamine content with agmatine to determine if attrition by cell death contributes to the growth-suppressive effects. We did not observe an increase in necrosis, DNA fragmentation, or chromatin condensation in Ha-Ras-transformed NIH-3T3 cells administered agmatine. In response to Ca2+-induced oxidative stress in kidney mitochondrial preparations, agmatine demonstrated attributes of a free radical scavenger by protecting against the oxidation of sulfhydryl groups and decreasing hydrogen peroxide content. The functional outcome was a protective effect against Ca2+-induced mitochondrial swelling and mitochondrial membrane potential collapse. We also observed decreased expression of proapoptotic Bcl-2 family members and of execution caspase-3, implying antiapoptotic potential. Indeed, we found that apoptosis induced by camptothecin or 5-fluorourocil was attenuated in cells administered agmatine. Agmatine may offer an alternative to the ornithine decarboxylase inhibitor difluoromethyl ornithine for depletion of intracellular polyamine content while avoiding the complications of increasing polyamine import and reducing the intracellular free radical scavenger capacity of polyamines. Depletion of intracellular polyamine content with agmatine suppressed cell growth, yet its antioxidant capacity afforded protection from mitochondrial insult and resistance to cellular apoptosis. These results could explain the beneficial outcomes observed with agmatine in models of injury and disease.

The zebrafish T-box genesno tailandspadetailare required for development of trunk and tail mesoderm and medial floor plate

Development ◽  
2002 ◽  
Vol 129 (14) ◽  
pp. 3311-3323 ◽  
Author(s):  
Sharon L. Amacher ◽  
Bruce W. Draper ◽  
Brian R. Summers ◽  
Charles B. Kimmel
Keyword(s):  
Embryonic Development ◽  
Neural Tube ◽  
Transcriptional Regulators ◽  
Floor Plate ◽  
Wild Type ◽  
T Box ◽  
No Tail ◽  
Ventral Neural Tube ◽  
Plate Formation ◽  
In Cells

T-box genes encode transcriptional regulators that control many aspects of embryonic development. Here, we demonstrate that the mesodermally expressed zebrafish spadetail (spt)/VegT and no tail (ntl)/Brachyury T-box genes are semi-redundantly and cell-autonomously required for formation of all trunk and tail mesoderm. Despite the lack of posterior mesoderm in spt–;ntl– embryos, dorsal-ventral neural tube patterning is relatively normal, with the notable exception that posterior medial floor plate is completely absent. This contrasts sharply with observations in single mutants, as mutations singly in ntl or spt enhance posterior medial floor plate development. We find that ntl function is required to repress medial floor plate and promote notochord fate in cells of the wild-type notochord domain and that spt and ntl together are required non cell-autonomously for medial floor plate formation, suggesting that an inducing signal present in wild-type mesoderm is lacking in spt–;ntl– embryos.

Ribosome Crystals in Necrotizing Cells from the Posterior Necrotic Zone of the Developing Chick Limb

1972 ◽  
Vol 11 (2) ◽  
pp. 403-414
Author(s):  
N. K. MOTTET ◽  
S. P. HAMMAR
Keyword(s):  
Electron Micrographs ◽  
Protein Crystals ◽  
Embryonic Chick ◽  
3 Dimensional ◽  
Surface Lattice ◽  
Plane Group ◽  
High Doses ◽  
In Cells

Ribosome and ‘protein crystals" were observed in electron micrographs of degenerating cells from the posterior necrotic zone (PNZ) of developing chick limbs, stages 22 through 24. Ribosome crystals were made up of basic units of 4 ribosomes arranged in a square tetramer. They were monomorphic with the surface lattice being of the p4 plane group. The 1-ribosome-thick sheets of crystals were often seen stacked upon one another forming a 3-dimensional P422 configuration. The percentage of crystallized ribosomes within degenerating PNZ cells is nearly directly proportional to the degree of degeneration of the cell. Crystals identical to this have been observed in numerous types of embryonic chick cells subjected to hypothermia. The ‘protein crystals’ were composed of straight filamentous structures separated by a distance of 20-30 nm. They were always closely associated with the ribosome crystals. These crystals are similar to those seen in cells treated with high doses of vinblastine and vincristine. The pathogenesis of the ribosome and protein crystals occurring in degenerating PNZ cells is unknown. They are not associated in any way with hypothermia or vinblastine-vincristine treatment.

scholarly journals Systematic Enzyme Mapping of Cellular Metabolism by Phasor-Analyzed Label-Free NAD(P)H Fluorescence Lifetime Imaging

2019 ◽  
Vol 20 (22) ◽  
pp. 5565 ◽  
Author(s):  
Leben ◽  
Köhler ◽  
Radbruch ◽  
Hauser ◽  
Niesner
Keyword(s):  
Fluorescence Lifetime ◽  
Metabolic Activity ◽  
Cellular Metabolism ◽  
Fluorescence Lifetime Imaging ◽  
Label Free ◽  
Analysis Framework ◽  
Cell Nuclei ◽  
Lifetime Imaging ◽  
Subcellular Resolution ◽  
In Cells

In the past years, cellular metabolism of the immune system experienced a revival, as it has become clear that it is not merely responsible for the cellular energy supply, but also impacts on many signaling pathways and, thus, on diverse cellular functions. Label-free fluorescence lifetime imaging of the ubiquitous coenzymes NADH and NADPH (NAD(P)H-FLIM) makes it possible to monitor cellular metabolism in living cells and tissues and has already been applied to study metabolic changes both under physiologic and pathologic conditions. However, due to the complex distribution of NAD(P)H-dependent enzymes in cells, whose distribution continuously changes over time, a thorough interpretation of NAD(P)H-FLIM results, in particular, resolving the contribution of various enzymes to the overall metabolic activity, remains challenging. We developed a systematic framework based on angle similarities of the phase vectors and their length to analyze NAD(P)H-FLIM data of cells and tissues based on a generally valid reference system of highly abundant NAD(P)H-dependent enzymes in cells. By using our analysis framework, we retrieve information not only about the overall metabolic activity, i.e., the fraction of free to enzyme-bound NAD(P)H, but also identified the enzymes predominantly active within the sample at a certain time point with subcellular resolution. We verified the performance of the approach by applying NAD(P)H-FLIM on a stromal-like cell line and identified a different group of enzymes that were active in the cell nuclei as compared to the cytoplasm. As the systematic phasor-based analysis framework of label-free NAD(P)H-FLIM can be applied both in vitro and in vivo, it retains the unique power to enable dynamic enzyme-based metabolic investigations, at subcellular resolution, in genuine environments.

scholarly journals Cyclin B3 activates the Anaphase-Promoting Complex/Cyclosome in meiosis and mitosis

2020 ◽  
Author(s):  
Damien Garrido ◽  
Mohammed Bourouh ◽  
Éric Bonneil ◽  
Pierre Thibault ◽  
Andrew Swan ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Chromosome Segregation ◽  
Ubiquitin Ligase ◽  
Anaphase Promoting Complex ◽  
Cells In Culture ◽  
Cyclin B3 ◽  
Mitosis And Meiosis ◽  
In Cells

ABSTRACTIn mitosis and meiosis, chromosome segregation is triggered by the Anaphase-Promoting Complex/Cyclosome (APC/C), a multi-subunit ubiquitin ligase that targets proteins for degradation, leading to the separation of chromatids. APC/C activation requires phosphorylation of its APC3 and APC1 subunits, which allows the APC/C to bind its Cdc20 co-activator. The identity of the kinase(s) responsible for APC/C activation in vivo is unclear. Cyclin B3 is required for meiotic anaphase in flies, worms and vertebrates, but whether it activates the APC/C is unclear. We found that Drosophila Cyclin B3 (CycB3) collaborates with PP2A-B55/Tws in embryonic development, indicating that CycB3 also promotes anaphase in mitosis. Moreover, CycB3 promotes APC/C activity and anaphase in cells in culture. We show that CycB3 physically associates with the APC/C, is required for phosphorylation of APC3, and promotes APC/C association with its co-activators. We propose that CycB3-Cdk1 directly phosphorylates the APC/C to activate it in both meiosis and mitosis.

6 CLONING AND EXPRESSION OF BOVINE FACTOR IN THE GERMLINE ALPHA (FIGLA) IN OOCYTES AND EARLY EMBRYOS: A POTENTIAL TARGET OF microRNA-212

2011 ◽  
Vol 23 (1) ◽  
pp. 109 ◽  
Author(s):  
S. K. Tripurani ◽  
K. B. Lee ◽  
G. W. Smith ◽  
J. Yao
Keyword(s):  
Embryonic Development ◽  
Expression Analysis ◽  
Transcriptional Regulator ◽  
Early Embryogenesis ◽  
Luciferase Reporter ◽  
Bovine Embryos ◽  
Cell Stage ◽  
Primordial Follicles ◽  
Maternal Transcript ◽  
In Cells

Factor In the GermLine Alpha (FIGLA), a basic helix-loop-helix transcription factor, was first identified in regulating coordinate expression of zona pellucida genes in mice. It plays a crucial role in the formation of primordial follicles and lack of FIGLA in mice alters the expression of many oocyte specific genes that are required for fertilization and early embryonic survival. The objective of this study was to characterise the expression and regulation of bovine FIGLA during early embryogenesis. The cloned bovine FIGLA cDNA is 660 bp in length, which encodes a protein of 165 amino acids. Expression of bovine FIGLA mRNA is restricted to ovarian tissue and can be detected in fetal ovaries harvested as early as 90 days of gestation when primordial follicles start to form. Expression analysis demonstrated that FIGLA mRNA is abundant in germinal vesicle and metaphase II stage oocytes, as well as in embryos from pronuclear to 8-cell stage, but barely detectable in embryos collected at morula and blastocyst stages, suggesting that FIGLA might be a maternal effect gene. Recent studies in zebrafish have highlighted the importance of non-coding small RNAs (microRNAs) as key regulatory molecules targeting maternal mRNAs for degradation during embryonic development. We hypothesised that FIGLA, as a maternal transcript, is regulated by microRNAs during early embryogenesis. Using microInspector, an algorithm for detection of possible interactions between microRNAs and target mRNA sequences, a microRNA binding site (miR-212) was identified in the 3′-UTR of the bovine FIGLA mRNA. Alignment of the 3′-UTR of FIGLA mRNAs from bovine, human and mouse shows complete conservation of the ‘seed’ region indicating that miR-212 might be a post-transcriptional regulator of FIGLA and the microRNA: mRNA interaction is evolutionary conserved. Expression analysis indicates that bovine miR-212 is expressed in oocytes and tends to increase at the 4-cell and 8-cell stage embryos followed by a decline at morula and blastocyst stages, indicating that miR-212 is presumably of maternal origin and potentially involved in maternal transcript degradation during the maternal-to-embryonic transition. To validate the role of miR-212 in silencing FIGLA, a luciferase reporter assay was performed using HeLa cells. The luciferase activity in cells expressing a luciferase construct containing the entire 3′ UTR of bovine FIGLA was suppressed by ∼40% in the presence of miR-212. We also investigated the stability of FIGLA mRNA in cells transfected with bovine FIGLA expression plasmid in the presence or absence of miR-212. Expression of bovine FIGLA mRNA was significantly reduced in the presence of mir-212 compared to control cells transfected with FIGLA construct alone. In summary, our data establish miR-212 as a potential post-transcriptional regulator of FIGLA during the maternal-to-embryonic transition in bovine embryos. Future studies aim to determine if miR-212 mimic can inhibit endogenous FIGLA expression in bovine embryos and its effect on subsequent embryonic development.

GROWTH OF ENTAMOEBA INVADENS IN ORGANOTYPIC CULTURES OF EMBRYONIC CHICK INTESTINE

1961 ◽  
Vol 7 (5) ◽  
pp. 685-695 ◽  
Author(s):  
E. Meerovitch
Keyword(s):  
Metabolic Activity ◽  
Embryonic Chick ◽  
Mucous Secretion ◽  
Organotypic Cultures ◽  
Fluid Medium ◽  
Intact Tissue ◽  
Entamoeba Invadens

Entamoeba invadens from axenic and monoxenic cultures was inoculated into expiants of embryonic chick intestine, which were then cultured in perfusion chambers at 30 °C. The growth and metabolic activity of the explants in cultures were evaluated in terms of fibroblastic outgrowth and extent of liquefaction of the plasma clots in which they were embedded. The effects of several media used to fill the perfusion chambers on the survival of the explants were studied. It was found that amoebae developed best in those explants which themselves showed most vitality; this was in turn related to the kind of fluid medium used in the culture. Amoebae in the explants fed on mucous secretion and on dead cells and penetrated into intact tissue without apparent histolytic activity. It is suggested that the living explants provided the amoebae with certain enzymes which the latter were unable to produce at the temperature of incubation. Approximately 40% of all cultures made became positive for amoebae. This is attributed to the fact that not all explants retained the amoebae injected into them, before they were placed in culture.

Nanodiamonds Interfere with Wnt-Regulated Cell Migration and Adipocyte Differentiation in Cells and Embryonic Development In Vivo

2016 ◽  
Vol 34 (1) ◽  
pp. 1600208 ◽  
Author(s):  
Hongyang Yi ◽  
Xiaojiao Li ◽  
Zhuyao Wang ◽  
Min Yin ◽  
Lihua Wang ◽  
...  
Keyword(s):  
Cell Migration ◽  
Embryonic Development ◽  
Adipocyte Differentiation ◽  
In Cells
Sign in / Sign up

Export Citation Format

Share Document