scholarly journals Urinary Exosomes

2009 ◽  
Vol 9 ◽  
pp. 1107-1118 ◽  
Author(s):  
Irena Dimov ◽  
Ljubinka Jankovic Velickovic ◽  
Vladisav Stefanovic
Keyword(s):  
Large Scale ◽  
Multivesicular Body ◽  
Cell Types ◽  
Response To Therapy ◽  
Tissue Samples ◽  
Disease Evolution ◽  
Urinary Exosomes ◽  
Cellular Source ◽  
Almost All

Exosomes are nanovesicles of endocytic origin that are secreted into the extracellular space or body fluids when a multivesicular body (MVB) fuses with the cell membrane. Interest in exosomes intensified after their description in antigen-presenting cells and the observation that they can significantly moderate immune responsesin vivo. In the past few years, several groups have reported on the secretion of exosomes by almost all cell types in an organism. In addition to a common set of membrane and cytosolic molecules, exosomes harbor unique subsets of proteins, reflecting their cellular source. Major research efforts were put into their surprisingly various biological functions and in translating knowledge into clinical practice. Urine provides an exciting noninvasive alternative to blood or tissue samples as a potential source of disease biomarkers. Urinary exosomes (UE) became the subject of serious studies just a few years ago. A recent large-scale proteomics-based study of normal UE revealed a myriad of proteins, including disease-related gene products. Thus, UE have valuable potential as a source of biomarkers for early detection of various types of diseases, monitoring the disease evolution and/or response to therapy. As a relatively new field of research, it still faces many challenges, but UE have already shown some straightforward potential.

scholarly journals Transcriptional and morphological profiling of parvalbumin interneuron subpopulations in the mouse hippocampus

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Lin Que ◽  
David Lukacsovich ◽  
Wenshu Luo ◽  
Csaba Földy
Keyword(s):  
Large Scale ◽  
Cell Types ◽  
Rna Seq ◽  
Neuronal Identity ◽  
Parvalbumin Interneurons ◽  
Different Types ◽  
Parvalbumin Interneuron ◽  
Cam Profile ◽  
Developmental Domains

AbstractThe diversity reflected by >100 different neural cell types fundamentally contributes to brain function and a central idea is that neuronal identity can be inferred from genetic information. Recent large-scale transcriptomic assays seem to confirm this hypothesis, but a lack of morphological information has limited the identification of several known cell types. In this study, we used single-cell RNA-seq in morphologically identified parvalbumin interneurons (PV-INs), and studied their transcriptomic states in the morphological, physiological, and developmental domains. Overall, we find high transcriptomic similarity among PV-INs, with few genes showing divergent expression between morphologically different types. Furthermore, PV-INs show a uniform synaptic cell adhesion molecule (CAM) profile, suggesting that CAM expression in mature PV cells does not reflect wiring specificity after development. Together, our results suggest that while PV-INs differ in anatomy and in vivo activity, their continuous transcriptomic and homogenous biophysical landscapes are not predictive of these distinct identities.

scholarly journals Postprandial transfer of colostral extracellular vesicles and their protein and miRNA cargo in neonatal calves

2019 ◽  
Author(s):  
Benedikt Kirchner ◽  
Dominik Buschmann ◽  
Vijay Paul ◽  
Michael W. Pfaffl
Keyword(s):  
Extracellular Vesicles ◽  
Expression Profiles ◽  
Bovine Milk ◽  
Cell Types ◽  
Specific Protein ◽  
In Vivo Experiments ◽  
Neonatal Calves ◽  
Almost All

Abstract Background Extracellular vesicles (EVs) such as exosomes are key regulators of intercellular communication that can be found in almost all bio fluids. Although studies in the last decade have made great headway in discerning the role of EVs in many physiological and pathophysiological processes, the bioavailability and impact of dietary EVs and their cargo still remain to be elucidated. Due to its widespread consumption and high content of EV-associated microRNAs and proteins, a major focus in this field has been set on EVs in bovine milk and colostrum. Despite promising in vitro studies in recent years that show high resiliency of milk EVs to degradation and uptake of milk EV cargo in a variety of intestinal and blood cell types, in vivo experiments continue to be inconclusive and sometimes outright contradictive. Results To resolve this discrepancy, we assessed the potential postprandial transfer of colostral EVs to the circulation of newborn calves by analysing colostrum-specific protein and miRNAs, including specific isoforms (isomiRs) in cells, EV isolations and unfractionated samples from blood and colostrum. Our findings reveal distinct populations of EVs in colostrum and blood from cows that can be clearly separated by density, particle concentration and protein content (BTN1A1, MFGE8). Postprandial blood samples of calves show a time-dependent increase in EVs that share morphological and protein characteristics of colostral EVs. Analysis of miRNA expression profiles by Next-Generation Sequencing gave a different picture however. Although significant postprandial expression changes could only be detected for calf EV samples, expression profiles show very limited overlap with highly expressed miRNAs in colostral EVs or colostrum in general. Conclusions Taken together our results indicate a selective uptake of membrane-associated protein cargo but not luminal miRNAs from colostral EVs into the circulation of neonatal calves.

scholarly journals Chandelier cell anatomy and function reveal a variably distributed but common signal

2020 ◽  
Author(s):  
Casey M. Schneider-Mizell ◽  
Agnes L. Bodor ◽  
Forrest Collman ◽  
Derrick Brittain ◽  
Adam A. Bleckert ◽  
...  
Keyword(s):  
Large Scale ◽  
Pyramidal Neurons ◽  
Cell Activity ◽  
Cortical Cell ◽  
Cell Types ◽  
Structural Features ◽  
Target Neuron ◽  
Chandelier Cell ◽  
Chandelier Cells

AbstractThe activity and connectivity of inhibitory cells has a profound impact on the operation of neuronal networks. While the average connectivity of many inhibitory cell types has been characterized, we still lack an understanding of how individual interneurons distribute their synapses onto their targets and how heterogeneous the inhibition is onto different individual excitatory neurons. Here, we use large-scale volumetric electron microscopy (EM) and functional imaging to address this question for chandelier cells in layer 2/3 of mouse visual cortex. Using dense morphological reconstructions from EM, we mapped the complete chandelier input onto 153 pyramidal neurons. We find that the number of input synapses is highly variable across the population, but the variability is correlated with structural features of the target neuron: soma depth, soma size, and the number of perisomatic synapses received. Functionally, we found that chandelier cell activity in vivo was highly correlated and tracks pupil diameter, a proxy for arousal state. We propose that chandelier cells provide a global signal whose strength is individually adjusted for each target neuron. This approach, combining comprehensive structural analysis with functional recordings of identified cell types, will be a powerful tool to uncover the wiring rules across the diversity of cortical cell types.

scholarly journals Nuclear RNA-seq of single neurons reveals molecular signatures of activation

2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Benjamin Lacar ◽  
Sara B. Linker ◽  
Baptiste N. Jaeger ◽  
Suguna Rani Krishnaswami ◽  
Jerika J. Barron ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Large Scale ◽  
Granule Cells ◽  
Mapk Pathway ◽  
Cell Types ◽  
Brain Regions ◽  
Activation Patterns ◽  
Environment Exposure

Abstract Single-cell sequencing methods have emerged as powerful tools for identification of heterogeneous cell types within defined brain regions. Application of single-cell techniques to study the transcriptome of activated neurons can offer insight into molecular dynamics associated with differential neuronal responses to a given experience. Through evaluation of common whole-cell and single-nuclei RNA-sequencing (snRNA-seq) methods, here we show that snRNA-seq faithfully recapitulates transcriptional patterns associated with experience-driven induction of activity, including immediate early genes (IEGs) such as Fos, Arc and Egr1. SnRNA-seq of mouse dentate granule cells reveals large-scale changes in the activated neuronal transcriptome after brief novel environment exposure, including induction of MAPK pathway genes. In addition, we observe a continuum of activation states, revealing a pseudotemporal pattern of activation from gene expression alone. In summary, snRNA-seq of activated neurons enables the examination of gene expression beyond IEGs, allowing for novel insights into neuronal activation patterns in vivo.

Excretion of urinary exosomal AQP2 in rats is regulated by vasopressin and urinary pH

2013 ◽  
Vol 305 (10) ◽  
pp. F1412-F1421 ◽  
Author(s):  
Yoshiki Higashijima ◽  
Hiroko Sonoda ◽  
Saki Takahashi ◽  
Hiroaki Kondo ◽  
Kanako Shigemura ◽  
...  
Keyword(s):  
Cell Types ◽  
Renal Secretion ◽  
Urinary Ph ◽  
Renal Epithelial Cell ◽  
Sodium Potassium ◽  
Urinary Exosomes ◽  
Urine Alkalinization ◽  
Apical Localization

Urinary exosomes are small vesicles secreted into urine from all renal epithelial cell types and known to contain proteins that are involved in renal secretion and reabsorption. Among these proteins, urinary exosomal aquaporin-2 (AQP2) has been suggested to be useful for diagnosis of renal disease. However, the mechanisms underlying the excretion of urinary exosomal AQP2 are largely unknown. In this study, we examined the mechanisms of urinary exosomal AQP2 excretion in vivo, using diuretics including furosemide (FS), an inhibitor of the sodium-potassium-chloride symporter; acetazolamide (ACTZ), an inhibitor of carbonic anhydrase; OPC-31260 (OPC), a vasopressin type 2 receptor antagonist; and NaHCO3, a urinary alkalizing agent. Samples of urine from rats were collected for 2 h just after treatment with each diuretic, and urinary exosomes were isolated by ultracentrifugation. Urinary exosomal AQP2 excretion was dramatically increased by treatment with FS accompanied by urine acidification or with ACTZ accompanied by urine alkalization. Immunohistochemistry showed that apical localization of AQP2 was clearly evident and the plasma vasopressin level was increased after each treatment. Although treatment with OPC alone had no significant effect, coadministration of OPC completely inhibited the FS-induced and partially reduced the ACTZ-induced responses, respectively. Treatment with NaHCO3 increased the excretion of urinary exosomal AQP2 accompanied by urine alkalization. This increased response was partially inhibited by coadministration of OPC. These data suggest that an increased plasma level of vasopressin promoted the excretion of urinary exosomal AQP2 and that urine alkalinization also increased it independently of vasopressin.

scholarly journals Microcarrier Screening and Evaluation for Dynamic Expansion of Human Periosteum-Derived Progenitor Cells in a Xenogeneic Free Medium

2021 ◽  
Vol 9 ◽  
Author(s):  
Kathleen Van Beylen ◽  
Ioannis Papantoniou ◽  
Jean-Marie Aerts
Keyword(s):  
Progenitor Cells ◽  
Cell Expansion ◽  
Progenitor Cell ◽  
Large Scale ◽  
Expansion Method ◽  
Cell Types ◽  
Free Medium ◽  
Efficient Expansion

An increasing need toward a more efficient expansion of adherent progenitor cell types arises with the advancements of cell therapy. The use of a dynamic expansion instead of a static planar expansion could be one way to tackle the challenges of expanding adherent cells at a large scale. Microcarriers are often reported as a biomaterial for culturing cells in suspension. However, the type of microcarrier has an effect on the cell expansion. In order to find an efficient expansion process for a specific adherent progenitor cell type, it is important to investigate the effect of the type of microcarrier on the cell expansion. Human periosteum-derived progenitor cells are extensively used in skeletal tissue engineering for the regeneration of bone defects. Therefore, we evaluated the use of different microcarriers on human periosteum-derived progenitor cells. In order to assess the potency, identity and viability of these cells after being cultured in the spinner flasks, this study performed several in vitro and in vivo analyses. The novelty of this work lies in the combination of screening different microcarriers for human periosteum-derived progenitor cells with in vivo assessments of the cells’ potency using the microcarrier that was selected as the most promising one. The results showed that expanding human periosteum-derived progenitor cells in spinner flasks using xeno-free medium and Star-Plus microcarriers, does not affect the potency, identity or viability of the cells. The potency of the cells was assured with an in vivo evaluation, where bone formation was achieved. In summary, this expansion method has the potential to be used for large scale cell expansion with clinical relevance.

scholarly journals An in vivo reporter for tracking lipid droplet dynamics in transparent zebrafish

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Dianne Lumaquin ◽  
Eleanor Johns ◽  
Emily Montal ◽  
Joshua M Weiss ◽  
David Ola ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Lipid Droplet ◽  
Lipid Droplets ◽  
Large Scale ◽  
Fluorescent Dyes ◽  
Cell Types ◽  
Structural Protein ◽  
Droplet Dynamics

Lipid droplets are lipid storage organelles found in nearly all cell types from adipocytes to cancer cells. Although increasingly implicated in disease, current methods to study lipid droplets in vertebrate models rely on static imaging or the use of fluorescent dyes, limiting investigation of their rapid in vivo dynamics. To address this, we created a lipid droplet transgenic reporter in whole animals and cell culture by fusing tdTOMATO to Perilipin-2 (PLIN2), a lipid droplet structural protein. Expression of this transgene in transparent casper zebrafish enabled in vivo imaging of adipose depots responsive to nutrient deprivation and high-fat diet. Simultaneously, we performed a large-scale in vitro chemical screen of 1280 compounds and identified several novel regulators of lipolysis in adipocytes. Using our Tg(-3.5ubb:plin2-tdTomato) zebrafish line, we validated several of these novel regulators and revealed an unexpected role for nitric oxide in modulating adipocyte lipid droplets. Similarly, we expressed the PLIN2-tdTOMATO transgene in melanoma cells and found that the nitric oxide pathway also regulated lipid droplets in cancer. This model offers a tractable imaging platform to study lipid droplets across cell types and disease contexts using chemical, dietary, or genetic perturbations.

scholarly journals Stem cells: an origin and marks

2020 ◽  
Vol 22 (2) ◽  
pp. 211-216
Author(s):  
A. V. Moskalev ◽  
B. Y. Gumilevskiy ◽  
A. V. Apchel ◽  
V. N. Cygan
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Molecular Marker ◽  
Population Level ◽  
Cell Types ◽  
Stem Cell Marker ◽  
Wide Range ◽  
Almost All

The basic physiological functions of stem cells are given: the ability to reproduce and generate offspring, which are manifested at the level of the population, and not of a single cell. The manifestation of these functions depends on the quantitative and qualitative composition of the microenvironment. Stem cells consist of two fundamentally different types: pluripotent, which exist only in vitro (in vitro) and tissue, existing in the postpartum body (in vivo). Stem cells can be replaced without limitation in vitro and lead to the appearance of a wide range of cell types. Tissue stem cells under normal conditions do not generate cells characteristic of other types of tissue. Stem cells include cells capable of expressing the gene products characteristic of them. However, there is no universal marker to differentiate stem cells from non-stem cells. A key marker of pluripotency is the transcription factor - a pituitary-specific transcription factor is positive. A component that can be found in almost all types of stem cells is the telomerase complex. Another stem cell marker is called CD34 glycoprotein. The functional activity of stem cells is associated with a molecular marker referred to as leucine-rich repeat containing G-protein bound to receptor 5. However, other types of cells do not express this marker. The physiological capabilities of stem cells depend both on the cells themselves and on their environment. The most reliable way to identify stem cells is to determine their phenotype in vivo. This suggests that stem cells do not carry a universal molecular marker. Most likely, they have significant differences from transplanted cells, and these differences cannot always be detected in individual cells, but only at the population level.

scholarly journals Mesenchymal Stem Cell-Derived Exosomes: Applications in Regenerative Medicine

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1959
Author(s):  
Mangesh D. Hade ◽  
Caitlin N. Suire ◽  
Zucai Suo
Keyword(s):  
Regenerative Medicine ◽  
Brain Injuries ◽  
Current Knowledge ◽  
Multivesicular Body ◽  
Cell Types ◽  
Biological Properties ◽  
Cellular Functions ◽  
Surface Receptors ◽  
Differentiation And Proliferation ◽  
Almost All

Exosomes are a type of extracellular vesicles, produced within multivesicular bodies, that are then released into the extracellular space through a merging of the multivesicular body with the plasma membrane. These vesicles are secreted by almost all cell types to aid in a vast array of cellular functions, including intercellular communication, cell differentiation and proliferation, angiogenesis, stress response, and immune signaling. This ability to contribute to several distinct processes is due to the complexity of exosomes, as they carry a multitude of signaling moieties, including proteins, lipids, cell surface receptors, enzymes, cytokines, transcription factors, and nucleic acids. The favorable biological properties of exosomes including biocompatibility, stability, low toxicity, and proficient exchange of molecular cargos make exosomes prime candidates for tissue engineering and regenerative medicine. Exploring the functions and molecular payloads of exosomes can facilitate tissue regeneration therapies and provide mechanistic insight into paracrine modulation of cellular activities. In this review, we summarize the current knowledge of exosome biogenesis, composition, and isolation methods. We also discuss emerging healing properties of exosomes and exosomal cargos, such as microRNAs, in brain injuries, cardiovascular disease, and COVID-19 amongst others. Overall, this review highlights the burgeoning roles and potential applications of exosomes in regenerative medicine.

scholarly journals An adhesion code ensures robust pattern formation during tissue morphogenesis

Science ◽  
2020 ◽  
Vol 370 (6512) ◽  
pp. 113-116 ◽  
Author(s):  
Tony Y.-C. Tsai ◽  
Mateusz Sikora ◽  
Peng Xia ◽  
Tugba Colak-Champollion ◽  
Holger Knaut ◽  
...  
Keyword(s):  
Large Scale ◽  
Ex Vivo ◽  
Cell Types ◽  
Neural Progenitors ◽  
Specific Cell ◽  
Adhesion Forces ◽  
Tissue Morphogenesis ◽  
Differential Adhesion ◽  
Mediate Cell

Animal development entails the organization of specific cell types in space and time, and spatial patterns must form in a robust manner. In the zebrafish spinal cord, neural progenitors form stereotypic patterns despite noisy morphogen signaling and large-scale cellular rearrangements during morphogenesis and growth. By directly measuring adhesion forces and preferences for three types of endogenous neural progenitors, we provide evidence for the differential adhesion model in which differences in intercellular adhesion mediate cell sorting. Cell type–specific combinatorial expression of different classes of cadherins (N-cadherin, cadherin 11, and protocadherin 19) results in homotypic preference ex vivo and patterning robustness in vivo. Furthermore, the differential adhesion code is regulated by the sonic hedgehog morphogen gradient. We propose that robust patterning during tissue morphogenesis results from interplay between adhesion-based self-organization and morphogen-directed patterning.

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