Evolution of mechanisms controlling epithelial morphogenesis across animals: new insights from dissociation-reaggregation experiments in the sponge Oscarella lobularis

Abstract Background The ancestral presence of epithelia in Metazoa is no longer debated. Porifera seem to be one of the best candidates to be the sister group to all other Metazoa. This makes them a key taxon to explore cell-adhesion evolution on animals. For this reason, several transcriptomic, genomic, histological, physiological and biochemical studies focused on sponge epithelia. Nevertheless, the complete and precise protein composition of cell–cell junctions and mechanisms that regulate epithelial morphogenetic processes still remain at the center of attention. Results To get insights into the early evolution of epithelial morphogenesis, we focused on morphogenic characteristics of the homoscleromorph sponge Oscarella lobularis. Homoscleromorpha are a sponge class with a typical basement membrane and adhaerens-like junctions unknown in other sponge classes. We took advantage of the dynamic context provided by cell dissociation-reaggregation experiments to explore morphogenetic processes in epithelial cells in a non-bilaterian lineage by combining fluorescent and electron microscopy observations and RNA sequencing approaches at key time-points of the dissociation and reaggregation processes. Conclusions Our results show that part of the molecular toolkit involved in the loss and restoration of epithelial features such as cell–cell and cell–matrix adhesion is conserved between Homoscleromorpha and Bilateria, suggesting their common role in the last common ancestor of animals. In addition, sponge-specific genes are differently expressed during the dissociation and reaggregation processes, calling for future functional characterization of these genes.

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Evolution of mechanisms controlling epithelial morphogenesis across animals: new insights from dissociation - reaggregation experiments in the sponge Oscarella lobularis.

10.1101/2021.03.22.436370 ◽

2021 ◽

Author(s):

Amelie Vernale ◽

Maria Mandela Prunster ◽

Fabio Marchiano ◽

Henry Debost ◽

Nicolas Brouilly ◽

...

Keyword(s):

Common Ancestor ◽

Functional Characterization ◽

Sister Group ◽

Epithelial Morphogenesis ◽

Cell Junctions ◽

Last Common Ancestor ◽

Cell Matrix ◽

Cell Matrix Adhesion ◽

Cell Cell

Background: The ancestral presence of epithelia in Metazoa is no longer debated. Even though Porifera seem to be the best candidates to be the sister group to all other Metazoa, hardly anything is known about the proteins involved in the composition of cell-cell junctions or about the mechanisms that regulate epithelial morphogenetic processes in this phylum. Results: To get insights into the early evolution of epithelial morphogenesis, we focused on morphogenic characteristics of the homoscleromorph sponge Oscarella lobularis. Homoscleromorpha are a sponge class with a typical basement membrane and adherens-like junctions unknown in other sponge classes. We took advantage of the dynamic context provided by cell dissociation-reaggregation experiments to explore morphogenetic processes in epithelial cells in an early lineage by combining fluorescent and electronic microscopy observations and RNA sequencing approaches at key time-points of the dissociation and reaggregation processes. Conclusions: Our results show that part of the molecular toolkit involved in the loss and restoration of epithelial features such as cell-cell and cell-matrix adhesion is conserved between Homoscleromorpha and Bilateria, suggesting their common role in the last common ancestor of animals. In addition, Sponge-specific genes are differently expressed during the dissociation and reaggregation processes, calling for future functional characterization of these genes.

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Evolution of mechanisms controlling epithelial morphogenesis across animals: new insights from dissociation - reaggregation experiments in the sponge Oscarella lobularis

10.21203/rs.3.rs-376467/v1 ◽

2021 ◽

Author(s):

Amélie Vernale ◽

Maria Mandela Prünster ◽

Fabio Marchianò ◽

Henry Debost ◽

Nicolas Brouilly ◽

...

Keyword(s):

Common Ancestor ◽

Functional Characterization ◽

Sister Group ◽

Epithelial Morphogenesis ◽

Cell Junctions ◽

Last Common Ancestor ◽

Cell Matrix ◽

Cell Matrix Adhesion ◽

Cell Cell

Abstract Background: The ancestral presence of epithelia in Metazoa is no longer debated. Even though Porifera seem to be the best candidates to be the sister group to all other Metazoa, hardly anything is known about the proteins involved in the composition of cell-cell junctions or about the mechanisms that regulate epithelial morphogenetic processes in this phylum. Results: To get insights into the early evolution of epithelial morphogenesis, we focused on morphogenic characteristics of the homoscleromorph sponge Oscarella lobularis. Homoscleromorpha are a sponge class with a typical basement membrane and adherens-like junctions unknown in other sponge classes. We took advantage of the dynamic context provided by cell dissociation-reaggregation experiments to explore morphogenetic processes in epithelial cells in an early lineage by combining fluorescent and electronic microscopy observations and RNA sequencing approaches at key time-points of the dissociation and reaggregation processes. Conclusions: Our results show that part of the molecular toolkit involved in the loss and restoration of epithelial features such as cell-cell and cell-matrix adhesion is conserved between Homoscleromorpha and Bilateria, suggesting their common role in the last common ancestor of animals. In addition, Sponge-specific genes are differently expressed during the dissociation and reaggregation processes, calling for future functional characterization of these genes.

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Epithelial Morphogenesis Driven by Cell-Matrix vs. Cell-Cell Adhesion

10.1101/2020.06.24.165795 ◽

2020 ◽

Author(s):

Shaohe Wang ◽

Kazue Matsumoto ◽

Kenneth M. Yamada

Keyword(s):

Cell Adhesion ◽

Single Cell ◽

Cell Sheet ◽

Branching Morphogenesis ◽

Epithelial Morphogenesis ◽

Epithelial Surface ◽

Matrix Adhesion ◽

Cell Matrix ◽

Cell Matrix Adhesion ◽

Cell Cell

SUMMARYMany embryonic organs undergo epithelial morphogenesis to form tree-like hierarchical structures. However, it remains unclear what drives the budding and branching of stratified epithelia, such as in embryonic salivary gland and pancreas. Here, we performed live-organ imaging of mouse embryonic salivary glands at single-cell resolution to reveal that budding morphogenesis is driven by expansion and folding of a distinct epithelial surface cell sheet characterized by strong cell-matrix adhesions and weak cell-cell adhesions. Profiling of single-cell transcriptomes of this epithelium revealed spatial patterns of transcription underlying these cell adhesion differences. We then synthetically reconstituted budding morphogenesis by experimentally suppressing E-cadherin expression and inducing basement membrane formation in 3D spheroid cultures of engineered cells, which required β1 integrin-mediated cell-matrix adhesion for successful budding. Thus, stratified epithelial budding, the key first step of branching morphogenesis, is driven by an overall combination of strong cell-matrix adhesion and weak cell-cell adhesion by peripheral epithelial cells.

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Epithelial Morphogenesis Driven by Cell-Matrix vs. Cell-Cell Adhesion

SSRN Electronic Journal ◽

10.2139/ssrn.3733158 ◽

2020 ◽

Author(s):

Shaohe Wang ◽

Kazue Matsumoto ◽

Kenneth M. Yamada

Keyword(s):

Cell Adhesion ◽

Epithelial Morphogenesis ◽

Cell Matrix ◽

Cell Cell

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Spatially Selective Imaging of Cell‐Matrix and Cell‐Cell Junctions by Electrochemiluminescence

Angewandte Chemie International Edition ◽

10.1002/anie.202101467 ◽

2021 ◽

Author(s):

Hao Ding ◽

Ping Zhou ◽

Wenxuan Fu ◽

Lurong Ding ◽

Weiliang Guo ◽

...

Keyword(s):

Cell Junctions ◽

Cell Matrix ◽

Cell Cell

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HGF Converts ErbB2/Neu Epithelial Morphogenesis to Cell Invasion

Molecular Biology of the Cell ◽

10.1091/mbc.e04-07-0567 ◽

2005 ◽

Vol 16 (2) ◽

pp. 550-561 ◽

Cited By ~ 77

Author(s):

Hanane Khoury ◽

Monica A. Naujokas ◽

Dongmei Zuo ◽

Veena Sangwan ◽

Melanie M. Frigault ◽

...

Keyword(s):

Growth Factor ◽

Hepatocyte Growth Factor ◽

Cell Invasion ◽

Single Cells ◽

Epithelial Morphogenesis ◽

Cell Junctions ◽

Junction Protein ◽

Hepatocyte Growth ◽

E Cadherin ◽

Cell Cell

Activation of the hepatocyte growth factor receptor Met induces a morphogenic response and stimulates the formation of branching tubules by Madin-Darby canine kidney (MDCK) epithelial cells in three-dimensional cultures. A constitutively activated ErbB2/Neu receptor, NeuNT, promotes a similar invasive morphogenic program in MDCK cells. Because both receptors are expressed in breast epithelia, are associated with poor prognosis, and hepatocyte growth factor (HGF) is expressed in stroma, we examined the consequence of cooperation between these signals. We show that HGF disrupts NeuNT-induced epithelial morphogenesis, stimulating the breakdown of cell-cell junctions, dispersal, and invasion of single cells. This correlates with a decrease in junctional proteins claudin-1 and E-cadherin, in addition to the internalization of the tight junction protein ZO-1. HGF-induced invasion of NT-expressing cells is abrogated by pretreatment with a pharmacological inhibitor of the mitogen-activated protein kinase kinase (MEK) pathway, which restores E-cadherin and ZO-1 at cell-cell junctions, establishing the involvement of MEK-dependent pathways in this process. These results demonstrate that physiological signals downstream from the HGF/Met receptor synergize with ErbB2/Neu to enhance the malignant phenotype, promoting the breakdown of cell-cell junctions and enhanced cell invasion. This is particularly important for cancers where ErbB2/Neu is overexpressed and HGF is a physiological growth factor found in the stroma.

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Ezrin regulates cell-cell and cell-matrix adhesion, a possible role with E-cadherin/beta-catenin

Journal of Cell Science ◽

10.1242/jcs.112.18.3081 ◽

1999 ◽

Vol 112 (18) ◽

pp. 3081-3090 ◽

Cited By ~ 4

Author(s):

S. Hiscox ◽

W.G. Jiang

Keyword(s):

Cancer Cells ◽

Focal Adhesions ◽

Beta Catenin ◽

Matrix Adhesion ◽

Cell Matrix ◽

Ezrin Expression ◽

Coated Surfaces ◽

Cell Matrix Adhesion ◽

E Cadherin ◽

Cell Cell

Ezrin, radixin, moesin and merlin form a subfamily of conserved proteins in the band 4.1 superfamily. The function of these proteins is to link the plasma membrane to the actin cytoskeleton. Merlin is defective or absent in schwannomas and meningiomas and has been suggested to function as a tumour suppressor. In this study, we have examined the role of ezrin as a potential regulator of the adhesive and invasive behaviour of tumour cells. We have shown that following inhibition of ezrin expression in colo-rectal cancer cells using antisense oligonucleotides, these cells displayed a reduced cell-cell adhesiveness together with a gain in their motile and invasive behaviour. These cells also displayed increased spreading over matrix-coated surfaces. Immunofluorescence studies revealed that antisense-treated cells also displayed an increased staining of paxillin in areas representing focal adhesions. Furthermore, coprecipitation studies revealed an association of ezrin with E-cadherin and beta-catenin. Induction of the phosphorylation of ezrin by orthovanadate and hepatocyte growth factor/scatter factor resulted in changes similar to those seen with antisense treatment, together with a marked decrease in the association of ezrin with both beta-catenin and E-cadherin. It is concluded that ezrin regulates cell-cell and cell-matrix adhesion, by interacting with cell adhesion molecules E-cadherin and beta-catenin, and may thus play an important role in the control of adhesion and invasiveness of cancer cells.

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Long-range and selective autoregulation of cell-cell or cell-matrix adhesions by cadherin or integrin ligands

Journal of Cell Science ◽

10.1242/jcs.111.3.347 ◽

1998 ◽

Vol 111 (3) ◽

pp. 347-357 ◽

Cited By ~ 10

Author(s):

S. Levenberg ◽

B.Z. Katz ◽

K.M. Yamada ◽

B. Geiger

Keyword(s):

Cell Surface ◽

Tyrosine Phosphorylation ◽

Long Range ◽

Cell Junctions ◽

Complete Suppression ◽

Beta Catenin ◽

Focal Contact ◽

Cell Matrix ◽

Immunofluorescence Labeling ◽

Cell Cell

In this study we demonstrate that local stimulation of cell surface cadherins or integrins induces a selective enhancement of adherens junction or focal contact assembly, respectively, throughout the cell. N-cadherin transfected CHO cells (CHO-Ncad) were incubated with different ligands including N-cadherin extracellular domain (NEC), anti-N-cadherin antibodies, fibronectin and concanavalin A (ConA), conjugated to synthetic beads. Electron microscopic examination indicated that both cadherin- and integrin-reactive beads bound tightly to the cell surface and were apparently endocytosed after several hours of incubation. The ConA-beads remained largely at the cell surface. Immunofluorescence labeling of the cells with antibodies to different adhesion-associated molecules indicated that both NEC- and anti-N-cadherin-conjugated beads induced a major increase in the level of junction-associated cadherin and beta-catenin labeling and a modest increase in junctional vinculin labeling, compared to untreated cells or cells bound to ConA-beads. FN-conjugated beads, on the other hand, significantly enhanced vinculin labeling at focal contacts and suppressed cadherin and beta-catenin staining in cell-cell junctions. The cadherin-reactive beads specifically stimulated tyrosine phosphorylation at cell-cell junctions, while the FN-beads increased the levels of focal contact-associated phosphotyrosine, as shown by immunofluorescence labeling of the cells for phosphotyrosine. Inhibition of this phosphorylation by genistein resulted in a complete suppression of the effects of both types of beads. These findings indicate that specific cadherin- and integrin-mediated surface interactions can trigger positively cooperative long-range signaling events which lead to the selective assembly of cell-cell or cell-matrix adhesions, and that these signals involve tyrosine phosphorylation.

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