Isolation and characterisation of watermelon (Citrullus lanatus) extracellular vesicles and their cargo

Mapping Intimacies ◽

10.1101/791111 ◽

2019 ◽

Author(s):

Kate Timms ◽

Beth Holder ◽

Anil Day ◽

John McLaughlin ◽

Melissa Westwood ◽

...

Keyword(s):

Extracellular Vesicles ◽

Citrullus Lanatus ◽

Cell Communication ◽

Functional Roles ◽

Cellular Debris ◽

Fruit Development And Ripening ◽

Crop Development ◽

Size And Morphology ◽

And Function ◽

Cell Cell

AbstractExtracellular vesicles (EVs) facilitate cell-cell communication in animals and are integral to many physiological and pathological processes. Evidence for the presence and function of EVs in plants is limited. Here, we report that EVs derived from watermelon fruit mesocarp are of similar size and morphology to the animal EV subtype known as exosomes. Analysis of EV constituents revealed that watermelon EVs are negative for endoplasmic reticulum markers, and that the miRNA and protein profiles differ from that of watermelon mesocarp cells, suggesting that these EVs are actively synthesised and are not merely cellular debris. Furthermore, we report a panel of proteins found in in watermelon EVs as well as the published proteomes of grape, grapefruit, lemon andArabidopsis thalianaEVs that are novel potential plant EV markers. Bioinformatic analyses suggest that plastids and multivesicular bodies are likely sites of biogenesis for EVs from watermelon and other plants. Predicted functional roles of watermelon EVs include development and metabolism, with several of their cargo molecules likely to be key in regulation of fruit development and ripening. Further understanding of how EVs may contribute to these processes would improve understanding of plant cell-cell communication and could aid in the harnessing of plant EVs for greater temporal control of crop development/ripening for the agricultural and retail industries.

Download Full-text

The Aquatic Invertebrate Hydra vulgaris Releases Molecular Messages Through Extracellular Vesicles

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.788117 ◽

2021 ◽

Vol 9 ◽

Author(s):

Maria Moros ◽

Eugenio Fergola ◽

Valentina Marchesano ◽

Margherita Mutarelli ◽

Giuseppina Tommasini ◽

...

Keyword(s):

Extracellular Vesicles ◽

Cell Communication ◽

Population Level ◽

Target Cells ◽

Chemical Information ◽

Hydra Vulgaris ◽

Physiological Processes ◽

Developmental Signalling ◽

And Function ◽

Cell Cell

Recent body of evidence demonstrates that extracellular vesicles (EVs) represent the first language of cell-cell communication emerged during evolution. In aquatic environments, transferring signals between cells by EVs offers protection against degradation, allowing delivering of chemical information in high local concentrations to the target cells. The packaging of multiple signals, including those of hydrophobic nature, ensures target cells to receive the same EV-conveyed messages, and the coordination of a variety of physiological processes across cells of a single organisms, or at the population level, i.e., mediating the population’s response to changing environmental conditions. Here, we purified EVs from the medium of the freshwater invertebrate Hydra vulgaris, and the molecular profiling by proteomic and transcriptomic analyses revealed multiple markers of the exosome EV subtype, from structural proteins to stress induced messages promoting cell survival. Moreover, positive and negative regulators of the Wnt/β-catenin signaling pathway, the major developmental pathway acting in body axial patterning, were identified. Functional analysis on amputated polyps revealed EV ability to modulate both head and foot regeneration, suggesting bioactivity of the EV cargo and opening new perspectives on the mechanisms of developmental signalling. Our results open the path to unravel EV biogenesis and function in all cnidarian species, tracing back the origin of the cell-cell, cross-species or cross-kingdom communication in aquatic ecosystems.

Download Full-text

Tracing back the origin of cell-cell communication: Hydra vulgaris releases extracellular vesicles delivering regulators of head and foot regeneration

10.21203/rs.3.rs-763240/v1 ◽

2021 ◽

Author(s):

Maria Moros ◽

Eugenio Fergola ◽

Valentina Marchesano ◽

Margherita Mutarelli ◽

Giuseppina Tommasini ◽

...

Keyword(s):

Extracellular Vesicles ◽

Cell Communication ◽

Population Level ◽

Active Role ◽

Target Cells ◽

Chemical Information ◽

Long Distance ◽

Hydra Vulgaris ◽

And Function ◽

Cell Cell

Abstract Recent body of evidence demonstrates that extracellular vesicles (EVs) represent the first language of cell-cell communication emerged during evolution. In aquatic environments, transferring signals between cells by EVs offer protection against degradation, allowing delivering of chemical information in high local concentrations to the target cells. The packaging of multiple signals, including those of hydrophobic nature, ensures target cells to receive the same EV-conveyed messages, and the coordination of a variety of physiological processes across cells of a single organisms, or at the population level, i.e. mediating the population´s response to changing environmental conditions. Here, we purified EVs from the medium of the freshwater invertebrate Hydra vulgaris, and the molecular profiling by proteomic and transcriptomic analyses revealed multiple markers of the exosome EV subtype. Moreover, positive and negative regulators of the Wnt/β-catenin signaling pathway, the major developmental pathway acting in body axial patterning, were identified. Functional analysis on amputated polyps revealed EV ability to interfere with both head and foot regeneration, suggesting an active role in setting up tissue gradients and oro-aboral polarity through delivery of short and long-distance signals. Our results open the path to unravel EV biogenesis and function in all cnidarian species, tracing back the origin of the cell-cell, cross-species or cross-kingdom communication in aquatic ecosystems

Download Full-text

Glycosylation of Cancer Extracellular Vesicles: Capture Strategies, Functional Roles and Potential Clinical Applications

Cells ◽

10.3390/cells10010109 ◽

2021 ◽

Vol 10 (1) ◽

pp. 109

Author(s):

Álvaro M. Martins ◽

Cátia C. Ramos ◽

Daniela Freitas ◽

Celso A. Reis

Keyword(s):

Extracellular Vesicles ◽

Biomarker Discovery ◽

De Novo ◽

Recipient Cell ◽

Cell Communication ◽

Cancer Biomarkers ◽

Functional Roles ◽

Molecular Information ◽

Glycosylation Pathway ◽

Organ Tropism

Glycans are major constituents of extracellular vesicles (EVs). Alterations in the glycosylation pathway are a common feature of cancer cells, which gives rise to de novo or increased synthesis of particular glycans. Therefore, glycans and glycoproteins have been widely used in the clinic as both stratification and prognosis cancer biomarkers. Interestingly, several of the known tumor-associated glycans have already been identified in cancer EVs, highlighting EV glycosylation as a potential source of circulating cancer biomarkers. These particles are crucial vehicles of cell–cell communication, being able to transfer molecular information and to modulate the recipient cell behavior. The presence of particular glycoconjugates has been described to be important for EV protein sorting, uptake and organ-tropism. Furthermore, specific EV glycans or glycoproteins have been described to be able to distinguish tumor EVs from benign EVs. In this review, the application of EV glycosylation in the development of novel EV detection and capture methodologies is discussed. In addition, we highlight the potential of EV glycosylation in the clinical setting for both cancer biomarker discovery and EV therapeutic delivery strategies.

Download Full-text

Are gap junctions necessary for cell-to-cell coupling of smooth muscle?: an update

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y92-063 ◽

1992 ◽

Vol 70 (4) ◽

pp. 481-490 ◽

Cited By ~ 22

Author(s):

R. E. Garfield ◽

G. Thilander ◽

M. G. Blennerhassett ◽

N. Sakai

Keyword(s):

Smooth Muscle ◽

Gap Junctions ◽

Current Knowledge ◽

Smooth Muscles ◽

Cell Communication ◽

Circular Muscle ◽

Cell Coupling ◽

And Function ◽

Longitudinal Layer ◽

Cell Cell

Earlier, it was questioned whether gap junctions (GJs) were necessary for cell–cell communication in smooth muscle, and GJs were not seen in some smooth muscles. We reexamined this question in the myometrium and in intestinal smooth muscle, in light of current knowledge of the presence and function of GJs. In the uterus, numerous studies show that an increase in GJ number is associated with the onset of delivery and is required for effective parturition. In all cases, this increase in GJ number and the changes in uterine contractility were correlated with increased electrical and metabolic coupling. Evidence for the much smaller, but detectable, degree of electrical coupling in the preterm uterus is explained by the small (but again detectable) number of GJs present. In the intestine, GJs are readily detected in the circular muscle layer but have not been described in the adjacent longitudinal layer. While our immunohistochemical studies failed to detect GJs in the longitudinal layer, this may not be adequate to prove their absence. Therefore, current knowledge of GJ number and function is adequate to explain cell–cell coupling in the uterus. Although it remains uncertain whether GJs are absent from the longitudinal muscle of the intestine, there is no definitive evidence that cell–cell coupling can occur by means other than GJs.Key words: gap junctions, myometrium, connexins, smooth muscle, cell communication.

Download Full-text

Cell-to-Cell Communication in Learning and Memory: From Neuro- and Glio-Transmission to Information Exchange Mediated by Extracellular Vesicles

International Journal of Molecular Sciences ◽

10.3390/ijms21010266 ◽

2019 ◽

Vol 21 (1) ◽

pp. 266 ◽

Cited By ~ 9

Author(s):

Gabriella Schiera ◽

Carlo Maria Di Liegro ◽

Italia Di Liegro

Keyword(s):

Learning And Memory ◽

Extracellular Vesicles ◽

Glial Cells ◽

Information Exchange ◽

System Development ◽

Cell Communication ◽

Nervous System Development ◽

The Body ◽

Pathological Conditions ◽

And Function

Most aspects of nervous system development and function rely on the continuous crosstalk between neurons and the variegated universe of non-neuronal cells surrounding them. The most extraordinary property of this cellular community is its ability to undergo adaptive modifications in response to environmental cues originating from inside or outside the body. Such ability, known as neuronal plasticity, allows long-lasting modifications of the strength, composition and efficacy of the connections between neurons, which constitutes the biochemical base for learning and memory. Nerve cells communicate with each other through both wiring (synaptic) and volume transmission of signals. It is by now clear that glial cells, and in particular astrocytes, also play critical roles in both modes by releasing different kinds of molecules (e.g., D-serine secreted by astrocytes). On the other hand, neurons produce factors that can regulate the activity of glial cells, including their ability to release regulatory molecules. In the last fifteen years it has been demonstrated that both neurons and glial cells release extracellular vesicles (EVs) of different kinds, both in physiologic and pathological conditions. Here we discuss the possible involvement of EVs in the events underlying learning and memory, in both physiologic and pathological conditions.

Download Full-text

Extracellular Vesicles Released From Articular Chondrocytes Play a Major Role in Cell–Cell Communication

Journal of Orthopaedic Research® ◽

10.1002/jor.24525 ◽

2019 ◽

Vol 38 (4) ◽

pp. 731-739 ◽

Cited By ~ 2

Author(s):

Xiaoming Liu ◽

Claire Shortt ◽

Fenglin Zhang ◽

Mariah Q. Bater ◽

Mary K. Cowman ◽

...

Keyword(s):

Extracellular Vesicles ◽

Articular Chondrocytes ◽

Cell Communication ◽

Cell Cell

Download Full-text

Inhibition of a v b 3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

10.21203/rs.3.rs-17483/v1 ◽

2020 ◽

Author(s):

Wanessa Altei ◽

Bianca Pachane ◽

Patty K. Santos ◽

Ligia Ribeiro ◽

Bong Hwan Sung ◽

...

Keyword(s):

Cancer Cells ◽

Extracellular Vesicles ◽

Cell Communication ◽

Organ Specific ◽

Breast Cells ◽

Mediate Cell ◽

First Time ◽

Cd63 Expression ◽

Cell Cell

Abstract Background: Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from cells and mediate cell-cell communication. Integrin adhesion receptors are enriched in small EVs (SEVs) and SEV-carried integrins have been shown to promote cancer cell migration and to mediate organ-specific metastasis; however, how integrins mediate these effects is not entirely clear and could represent a combination of EV binding to extracellular matrix and cells.Methods: To probe integrin role in EVs binding and uptake, we employed a disintegrin inhibitor (DisBa-01) of integrin binding with specificity for avb3 integrin. EVs were purified from MDA-MB-231 cells conditioned media by serial centrifugation method. Isolated EVs were characterized by different techniques and further employed in adhesion, uptake and co-culture experiments.Results: We find that SEVs secreted from MDA-MB-231 breast cancer cells carry avb3 integrin and bind directly to fibronectin-coated plates, which is inhibited by DisBa-01. SEV coating on tissue culture plates also induces adhesion of MDA-MB-231 cells, which is inhibited by DisBa-01 treatment. Analysis of EV uptake and interchange between cells reveals that the amount of CD63-positive EVs delivered from malignant MDA-MB-231 breast cells to non-malignant MCF10A breast epithelial cells is reduced by DisBa-01 treatment. Inhibition of avb3 integrin decreases CD63 expression in cancer cells suggesting an effect on SEV content.Conclusion: In summary, our findings demonstrate for the first time a key role of avb3 integrin in cell-cell communication through SEVs.

Download Full-text

Inhibition of a v b 3 integrin impairs adhesion and uptake of tumor-derived small extracellular vesicles

10.21203/rs.3.rs-17483/v2 ◽

2020 ◽

Author(s):

Wanessa Altei ◽

Bianca Pachane ◽

Patty K. Santos ◽

Ligia Ribeiro ◽

Bong Hwan Sung ◽

...

Keyword(s):

Cancer Cells ◽

Extracellular Vesicles ◽

Cell Communication ◽

Αvβ3 Integrin ◽

Organ Specific ◽

Breast Cells ◽

Mediate Cell ◽

First Time ◽

Cell Cell

Abstract Background: Extracellular vesicles (EVs) are lipid-bound particles that are naturally released from cells and mediate cell-cell communication. Integrin adhesion receptors are enriched in small EVs (SEVs) and SEV-carried integrins have been shown to promote cancer cell migration and to mediate organ-specific metastasis; however, how integrins mediate these effects is not entirely clear and could represent a combination of EV binding to extracellular matrix and cells. Methods: To probe integrin role in EVs binding and uptake, we employed a disintegrin inhibitor (DisBa-01) of integrin binding with specificity for αvβ3 integrin. EVs were purified from MDA-MB-231 cells conditioned media by serial centrifugation method. Isolated EVs were characterized by different techniques and further employed in adhesion, uptake and co-culture experiments. Results: We find that SEVs secreted from MDA-MB-231 breast cancer cells carry αvβ3 integrin and bind directly to fibronectin-coated plates, which is inhibited by DisBa-01. SEV coating on tissue culture plates also induces adhesion of MDA-MB-231 cells, which is inhibited by DisBa-01 treatment. Analysis of EV uptake and interchange between cells reveals that the amount of CD63-positive EVs delivered from malignant MDA-MB-231 breast cells to non-malignant MCF10A breast epithelial cells is reduced by DisBa-01 treatment. Inhibition of αvβ3 integrin decreases CD63 expression in cancer cells suggesting an effect on SEV content. Conclusion: In summary, our findings demonstrate for the first time a key role of αvβ3 integrin in cell-cell communication through SEVs.

Download Full-text