scholarly journals COMPARATIVE ANALYSIS OF THE CONCENTRATION OF INJECTED HORSERADISH PEROXIDASE IN CYTOPLASMIC GRANULES OF THE KIDNEY CORTEX, IN THE BLOOD, URINE, AND LIVER

1971 ◽  
Vol 48 (3) ◽  
pp. 620-632 ◽  
Author(s):  
Werner Straus
Keyword(s):  
Horseradish Peroxidase ◽  
Marked Decrease ◽  
Cell Types ◽  
Kidney Cortex ◽  
Renal Tubules ◽  
Protein Uptake ◽  
Equal Dose ◽  
Wide Range ◽  
Glomerular Filtrate ◽  
Load Dose

The concentration of horseradish peroxidase in total particulate fractions from the kidney cortex did not change much during the first few hours after injection, as long as most of the injected protein was not yet cleared from the blood. It decreased at a rate of 6–8% per hr afterwards. The concentration of peroxidase in total particulate fractions increased in proportion to the load (dose) over a wide range, suggesting that a constant fraction of the protein was reabsorbed by micropinocytic vesicles into the tubule cells from the glomerular filtrate. The amount of peroxidase excreted in the urine also increased in proportion to the injected dose. The proportion of peroxidase taken up by the liver, however, decreased several times when the dose was increased. A marked decrease of protein uptake into the kidney cortex and an increase of urinary excretion were observed when rats received a second, equal dose of peroxidase 4 hr after the first injection, and the rate of clearance of peroxidase from the blood was decreased after the second injection. The liver, on the other hand, took up almost twice as much peroxidase after two injections as after one. The uptake of peroxidase by the kidney cortex increased with age. Cytochemical observations on the preferential absorption of peroxidase by certain cell types and segments of the renal tubules in relation to dose are reported.

scholarly journals Distribution of horseradish peroxidase (HRP)-anti-HRP immune complexes in mouse spleen with special reference to follicular dendritic cells.

1978 ◽  
Vol 79 (1) ◽  
pp. 184-199 ◽  
Author(s):  
L L Chen ◽  
A M Frank ◽  
J C Adams ◽  
R M Steinman
Keyword(s):  
Dendritic Cells ◽  
Horseradish Peroxidase ◽  
Immune Complexes ◽  
Cell Types ◽  
Mouse Spleen ◽  
Follicular Dendritic Cells ◽  
Quantitative Studies ◽  
Wide Range ◽  
Follicular Dendritic

The distribution of immune complexes has been studied in mouse spleen stimulated to contain many germinal centers (GC's). Horseradish peroxidase (HRP)-anti-HRP complexes were used as an appropriately precise and sensitive model. We were primarily interested in the relative abilities of three cell types to interact with complexes: lymphocytes, macrophages, and follicular dendritic cells (FDC's). The latter are distinctive, nonendocytic, stellate cells located primarily at the transition of mantle and GC zones of 2 degrees lymphoid follicles (Chen, L. L., J. C. Adams, and R. M. Steinman, 1978, J. Cell Biol. 77:148). Binding of immune complexes to lymphocytes could not be visualized in situ. Macrophages avidly interiorized complexes into lysosomes, but did not retain them extracellularly. In contrast, FDC's could retain HRP-anti-HRP extracellularly under appropriate conditions, but did not endocytose them. Cytochemical reactivity accumulated progressively on FDC's 1--6 h after administration of complexes i.v., remained stable in amount and location for 1 day, and then was progressively lost over a 1- to 5-day period. Several variables in the association of complexes with macrophages and FDC's were pursued. Only 1 microgram of complexed HRP had to be administered to visualize binding to both cell types. Macrophages interiorized complexes formed in a wide range of HRP/anti-HRP ratios, while FDC's associated with complexes formed in HRP excess only. Quantitative studies with [125I]HRP-anti-HRP demonstrated that 20% of the splenic load of HRP associated with FDC's. Complexes formed with an F(ab')2 anti-HRP were distributed primarily in macrophages. When the levels of the third component of serum complement were depleted by prior treatment with cobra venom factor, uptake of complexes by macrophages was reduced some 50% whereas association with FDC's was abolished. The fact that antigen excess complexes are retained extracellularly strengthens the idea that they are immunogenic. Finally, the association of complexes with FDC's seems to retard the entry of antigen into the GC proper.

Analysis of clonal restriction of cell mingling in Xenopus

1985 ◽  
Vol 312 (1153) ◽  
pp. 57-65 ◽  
Keyword(s):  
Horseradish Peroxidase ◽  
Cell Types ◽  
Cell Group ◽  
Cell Stage ◽  
Wide Range ◽  
Cell Groups ◽  
Cell Layers ◽  
Single Blastomeres

Cell fates were traced by injecting horseradish peroxidase into single blastomeres of Xenopus embryos at 2- to 512-cell stages. At later stages the number, types and locations of all labelled progeny were observed. Progeny of a single labelled ancestral cell divided coherently until the 12th cell generation, the onset of gastrulation, and then dispersed and mingled with unlabelled cells. Cell mingling was restricted at mediolateral and anterior—posterior boundaries. These boundaries were always respected by progeny of any blastomere labelled at the 512-cell stage but they were frequently crossed by progeny of blastomeres labelled at the 256-cell or earlier stages. The boundaries defined seven morphological compartments each populated exclusively by a group of ancestral cells at the 512-cell stage. Each blastomere that contributed progeny to the nervous system also gave rise to a wide range of cell types in all three primary germ cell layers but the clone was restricted to a single compartment. Analysis of clonal restriction of cell mingling was done in vitro . Twenty to thirty blastomeres were excised from one ancestral cell group at the 512-cell stage and combined in vitro with 20-30 blastomeres from another group. One group of blastomeres labelled with horseradish peroxidase was placed in contact with another group of unlabelled blastomeres, maintained in vitro for up to 2 days, and then processed histologically to show the distribution of labelled and unlabelled cells. Mingling was significantly greater in combinations of two of the same ancestral cell groups than in combinations of two different ancestral cell groups. A similar result was observed when a single labelled cell was combined with either the same or different ancestral cells. In all experiments the cells were significantly larger in combinations of different ancestral cell groups, indicating that they had undergone fewer divisions. These results are consistent with the hypothesis that boundaries observed in vivo are lines of clonal restriction formed by mutual inhibition of cell motility and cell division following contact between progeny of different ancestral cell groups.

scholarly journals The atlas of RNase H antisense oligonucleotide distribution and activity in the CNS of rodents and non-human primates following central administration

2020 ◽  
Author(s):  
Paymaan Jafar-nejad ◽  
Berit Powers ◽  
Armand Soriano ◽  
Hien Zhao ◽  
Daniel A Norris ◽  
...  
Keyword(s):  
Motor Neurons ◽  
Muscular Atrophy ◽  
Neurological Diseases ◽  
Cell Types ◽  
Rnase H ◽  
Central Administration ◽  
Spinal Motor Neurons ◽  
New Class ◽  
Wide Range ◽  
Therapeutic Development

Abstract Antisense oligonucleotides (ASOs) have emerged as a new class of drugs to treat a wide range of diseases, including neurological indications. Spinraza, an ASO that modulates splicing of SMN2 RNA, has shown profound disease modifying effects in Spinal Muscular Atrophy (SMA) patients, energizing efforts to develop ASOs for other neurological diseases. While SMA specifically affects spinal motor neurons, other neurological diseases affect different central nervous system (CNS) regions, neuronal and non-neuronal cells. Therefore, it is important to characterize ASO distribution and activity in all major CNS structures and cell types to have a better understanding of which neurological diseases are amenable to ASO therapy. Here we present for the first time the atlas of ASO distribution and activity in the CNS of mice, rats, and non-human primates (NHP), species commonly used in preclinical therapeutic development. Following central administration of an ASO to rodents, we observe widespread distribution and target RNA reduction throughout the CNS in neurons, oligodendrocytes, astrocytes and microglia. This is also the case in NHP, despite a larger CNS volume and more complex neuroarchitecture. Our results demonstrate that ASO drugs are well suited for treating a wide range of neurological diseases for which no effective treatments are available.

scholarly journals Energetics of mesoscale cell turbulence in two-dimensional monolayers

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Shao-Zhen Lin ◽  
Wu-Yang Zhang ◽  
Dapeng Bi ◽  
Bo Li ◽  
Xi-Qiao Feng
Keyword(s):  
Kinetic Energy ◽  
Tumor Metastasis ◽  
Cell Types ◽  
Boltzmann Distribution ◽  
Scaling Exponent ◽  
Biological Processes ◽  
Two Dimensional ◽  
Cell Monolayers ◽  
Wide Range ◽  
Epithelial Cell Monolayers

AbstractInvestigation of energy mechanisms at the collective cell scale is a challenge for understanding various biological processes, such as embryonic development and tumor metastasis. Here we investigate the energetics of self-sustained mesoscale turbulence in confluent two-dimensional (2D) cell monolayers. We find that the kinetic energy and enstrophy of collective cell flows in both epithelial and non-epithelial cell monolayers collapse to a family of probability density functions, which follow the q-Gaussian distribution rather than the Maxwell–Boltzmann distribution. The enstrophy scales linearly with the kinetic energy as the monolayer matures. The energy spectra exhibit a power-decaying law at large wavenumbers, with a scaling exponent markedly different from that in the classical 2D Kolmogorov–Kraichnan turbulence. These energetic features are demonstrated to be common for all cell types on various substrates with a wide range of stiffness. This study provides unique clues to understand active natures of cell population and tissues.

The Mode of Excretion of Ammonia and Urea in Xenopus Laevis

1961 ◽  
Vol 38 (4) ◽  
pp. 695-705
Author(s):  
J. B. BALINSKY ◽  
E. BALDWIN
Keyword(s):  
Xenopus Laevis ◽  
Blood Concentration ◽  
Concentration Ratio ◽  
Ammonia Excretion ◽  
Ammonia Concentration ◽  
Mean Value ◽  
Urea Excretion ◽  
Wide Range ◽  
Glomerular Filtrate ◽  
The Mean

1. Eighty-two single determinations of ammonia and urea excretion by Xenopus laevis indicated that the percentage of ammonia varied from 40 to 80%, with a mean value of 62%. 2. Measurements of excretion on successive days after feeding showed that a large amount of ammonia was produced soon after feeding, but that ammonia excretion declined rapidly. Urea excretion, not so high initially, remained more or less constant until the third or fourth day, often exceeding ammonia excretion at that time. Thereafter, it also declined and the excretion of both substances reached a constant starvation level by the fifteenth day. 3. Both ammonia and urea excretion were equally affected by temperature. The Q10's were near 2 in the range 20-30° C., but greater in the range 10-20° C. 4. At least 86% of ammonia, and 81% of urea were excreted through the cloaca. 5. The mean 24 hr. urine output of Xenopus at 20% C. was 23.6 ml. per 100 g. body weight. 6. Although the blood ammonia concentration did not appear to be zero, the urine/blood concentration ratio of ammonia was greater than 100. The urine/blood concentration ratio of urea was not significantly different from unity, and constant over a very wide range of concentrations. 7. The above result is interpreted to indicate passive glomerular filtration of urea, and little or no tubular reabsorption of water. 8. It is suggested that ammonia is formed in the kidney, and actively secreted into the glomerular filtrate.

Df31 is a novel nuclear protein involved in chromatin structure in Drosophila melanogaster

2001 ◽  
Vol 114 (1) ◽  
pp. 37-47 ◽  
Author(s):  
G. Crevel ◽  
H. Huikeshoven ◽  
S. Cotterill
Keyword(s):  
Cell Types ◽  
Drosophila Embryo ◽  
Cell Fractionation ◽  
General Function ◽  
Structural Protein ◽  
Chromosomal Structure ◽  
Binding Characteristics ◽  
Wide Range ◽  
Cellular Dna

We originally isolated the Df31 protein from Drosophila embryo extracts as a factor which could decondense Xenopus sperm, by removing the sperm specific proteins and interacting with histones to facilitate their loading onto DNA. We now believe that this protein has a more general function in cellular DNA metabolism. The Df31 gene encodes a very hydrophilic protein with a predicted molecular mass of 18.5 kDa. Immunostaining showed that Df31 was present in a wide range of cell types throughout differentiation and in both dividing and non-dividing cells. In all cases the protein is present in large amounts, comparable with the level of nucleosomes. Injection of antisense oligonucleotides to lower the level of Df31 in embryos caused severe disruption of the nuclear structure. Large irregular clumps of DNA were formed, and in most cases the amount of DNA associated with each clump was more than that found in a normal nucleus. Immunofluorescence, cell fractionation, and formaldehyde cross-linking show that Df31 is associated with chromatin and that a significant fraction of it binds very tightly. It also shows the same binding characteristics when loaded onto chromatin in vitro. Chromatin fractionation shows that Df31 is tightly associated with nucleosomes, preferentially with oligonucleosomes. Despite this no differences were observed in the properties of nucleosomes loaded in the in vitro system in the presence and absence of Df31. These results suggest that Df31 has a role in chromosomal structure, most likely acting as a structural protein at levels of folding higher than that of nucleosomes.

scholarly journals RNA-Seq Data-Mining Allows the Discovery of Two Long Non-Coding RNA Biomarkers of Viral Infection in Humans

2020 ◽  
Vol 21 (8) ◽  
pp. 2748 ◽  
Author(s):  
Ruth Barral-Arca ◽  
Alberto Gómez-Carballa ◽  
Miriam Cebey-López ◽  
María José Currás-Tuala ◽  
Sara Pischedda ◽  
...  
Keyword(s):  
Gene Expression ◽  
Viral Infections ◽  
Umbilical Vein ◽  
Cell Types ◽  
Dermal Fibroblasts ◽  
Learning Approaches ◽  
Rna Seq ◽  
Wide Range ◽  
Healthy Control ◽  
Umbilical Vein Endothelial Cells

There is a growing interest in unraveling gene expression mechanisms leading to viral host invasion and infection progression. Current findings reveal that long non-coding RNAs (lncRNAs) are implicated in the regulation of the immune system by influencing gene expression through a wide range of mechanisms. By mining whole-transcriptome shotgun sequencing (RNA-seq) data using machine learning approaches, we detected two lncRNAs (ENSG00000254680 and ENSG00000273149) that are downregulated in a wide range of viral infections and different cell types, including blood monocluclear cells, umbilical vein endothelial cells, and dermal fibroblasts. The efficiency of these two lncRNAs was positively validated in different viral phenotypic scenarios. These two lncRNAs showed a strong downregulation in virus-infected patients when compared to healthy control transcriptomes, indicating that these biomarkers are promising targets for infection diagnosis. To the best of our knowledge, this is the very first study using host lncRNAs biomarkers for the diagnosis of human viral infections.

Comparative physiology of insect renal function

1981 ◽  
Vol 241 (5) ◽  
pp. R241-R257 ◽  
Author(s):  
J. Phillips
Keyword(s):  
Surface Area ◽  
Total Surface Area ◽  
Malpighian Tubules ◽  
Thick Ascending Limb ◽  
Small Surface ◽  
Multiplier System ◽  
Wide Range ◽  
Glomerular Filtrate ◽  
Unit Body Weight ◽  
Anion Transfer

Transport mechanisms and their control in various segments of insect excretory systems are reviewed and compared to those of vertebrate nephrons, exocrine glands, and hindguts. Formation of the primary urine in most insect Malpighian tubules (MT) is by isosmotic secretion, which is driven by an apical cation (K+) pump rather than by Na+-K+-ATPase. Unlike the glomerular filtrate of vertebrates, insect MT fluid is very different from the blood in composition, often having very high K+-to-Na+ ratios, and urine-to-plasma values much less than unity for most other solutes. The total surface area of insect MT is some 20 times that of vertebrate glomeruli per unit body weight. Secretion of MT fluid is regulated by neuropeptides over a wide range of rats, similar to glomerular filtration rate values for many vertebrate kidneys. Several secretory mechanisms for selected solutes are probably common to insect and vertebrate tubules. Unlike vertebrates, insects usually reabsorb most of the filtered water, ions, and metabolites in the rectum, which has a small surface area relative to the MT. The rectum is also where ionic and osmotic composition of the excreta is finally adjusted, under the control of neuropeptide hormones. In the rectum, insect excreta can become as hyperosmotic as mammalian urine, even though a countercurrent multiplier system is not present. Active transport of Cl- predominates in both locust rectum and the thick ascending limb of Henle's loop, but the characteristics of the anion transfer process are quite different in these two epithelia.xs

The Florey Lecture, 1988 - From egg to embryo: the initiation of cell differentiation in Amphibia

1989 ◽  
Vol 237 (1286) ◽  
pp. 11-25 ◽  
Keyword(s):  
Muscle Cell ◽  
Gene Activation ◽  
Cell Formation ◽  
Gene Promoter ◽  
Cell Types ◽  
Specific Gene ◽  
Embryonic Induction ◽  
Differentiated Cells ◽  
Wide Range ◽  
Muscle Genes

Some of the principles by which different cell types first arise at the beginning of animal development are illustrated by muscle cell formation in Amphibia. If the nucleus of a differentiated muscle cell is transplanted to an enucleated egg, some of the resulting embryos develop into tadpoles with a wide range of normally differentiated cells. These experiments show that genes undergo major changes in activity as a response to components of egg cytoplasm. Two fundamental mechanisms account for the regional activation of genes in early embryos. One involves the effect of localized ‘determinants’ in egg cytoplasm, and the other concerns cell interactions or embryonic induction. Both these mechanisms seem to be responsible for muscle cell formation in amphibian development. The old problem of embryonic induction has recently become accessible to analysis at the molecular level, especially in the case of the mesoderm or muscle-forming induction. This has been greatly facilitated by using a sensitive and quantitative assay to detect the first transcripts of muscle genes a few hours after the start of induction. The role of early events and of interactions among like cells during response to induction is discussed. In analysing specific gene activation following induction, DNA injection into fertilized eggs has shown that a very small part of the cardiac actin gene promoter is sufficient to enable it to respond to induction. Although the experimental work summarized here has been done on amphibian embryos, which are more suitable than other embryos for embryological manipulation, the conclusions reached are believed to be generally applicable to the development of other organisms.

scholarly journals Mapping calcium dynamics in a developing tubular structure

2020 ◽  
Author(s):  
Jorgen Hoyer ◽  
Morsal Saba ◽  
Daniel Dondorp ◽  
Kushal Kolar ◽  
Riccardo Esposito ◽  
...  
Keyword(s):  
Cell Types ◽  
Feedback Mechanism ◽  
Adult Brain ◽  
Actin Dynamics ◽  
Cytoskeletal Network ◽  
Notochord Cell ◽  
Wide Range ◽  
Store Operated Calcium Entry ◽  
And Function ◽  
Cell Intercalation

AbstractCalcium is a ubiquitous and versatile second messenger that plays a central role in the development and function of a wide range of cell types, tissues and organs. Despite significant recent progress in the understanding of calcium (Ca2+) signalling in organs such as the developing and adult brain, we have relatively little knowledge of the contribution of Ca2+ to the development of tubes, structures widely present in multicellular organisms. Here we image Ca2+ dynamics in the developing notochord of Ciona intestinalis. We show that notochord cells exhibit distinct Ca2+ dynamics during specific morphogenetic events such as cell intercalation, cell elongation and tubulogenesis. We used an optogenetically controlled Ca2+ actuator to show that sequestration of Ca2+ results in defective notochord cell intercalation, and pharmacological inhibition to reveal that stretch-activated ion channels (SACs), inositol triphosphate receptor (IP3R) signalling, Store Operated Calcium Entry (SOCE), Sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) and gap junctions are required for regulating notochord Ca2+ activity during tubulogenesis. Cytoskeletal rearrangements drive the cell shape changes that accompany tubulogenesis. In line with this, we show that Ca2+ signalling modulates reorganization of the cytoskeletal network across the morphogenetic events leading up to and during tubulogenesis of the notochord. We additionally demonstrate that perturbation of the actin cytoskeleton drastically remodels Ca2+ dynamics, suggesting a feedback mechanism between actin dynamics and Ca2+ signalling during notochord development. This work provides a framework to quantitatively define how Ca2+ signalling regulates tubulogenesis using the notochord as model organ, a defining structure of all chordates.

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