β integrin-like protein-mediated adhesion and its disturbances during cell cultivation of the mussel Mytilus trossulus

Abstract

In this study, we focus on the specific contribution of β integrin-like protein to adhesion-mediated events in molluscan larval cells in culture that could not have been investigated within the whole animal. An analysis of disturbances to cell-substratum adhesion, caused by the integrin receptor inhibiting Arg-Gly-Asp-Ser (RGDS)-peptide, the Ca(2+)/Mg(2+)-chelators and the stress influence of freezing-thawing, reveals that all these factors resulted in the partial destruction of the integrin-extracellular matrix (ECM) interaction in culture and, in particular, changes in the distribution and relative abundance of β integrin-positive cells. The experiments, carried out on selected substrates, found that β integrin-positive cells demonstrate different affinities for the substrates. This finding further supports the assumption that epithelial differentiation in cultivated cells of larval Mytilus may be mediated by β integrin-like proteins via binding to laminin; direct binding to other components of the ECM could not be demonstrated. The mussel β integrin-positive cells are not involved in myogenic or neuronal differentiation on any of the substrates but part of them has tubulin-positive cilia, forming some epithelia-like structures. Our data indicate that β integrin-positive cells are able to proliferate in vitro which suggests that they could participate in renewing the digestive epithelium in larvae. The findings provide evidence that the distribution pattern of β integrin-like protein depends on the cell type and the factors influencing the adhesion.

The Members of the Highly Diverse Crassostrea gigas Integrin Family Cooperate for the Generation of Various Immune Responses

“…In humans, α5β1, α8β1, αVβ1, αVβ3, αVβ5, αVβ6, αVβ8, and αIIbβ3 belong to the RGD-binding receptors, and they can be involved in cellular immunity, cell motility, cell proliferation, and cell differentiation through the interaction with RGDCP ( 3 ). In invertebrates, the accumulating evidences have revealed that RGDCP can also bind hemocytes and inhibit the cell activities in Botryllus schlosseri ( 79 ), Pacifastacus leniusculus ( 26 ), Lymnaea stagnalis ( 39 ), Mytilus trossulus ( 41 ), etc., suggesting the existence of RGD-binding integrins in such animals. In the present study, Cg α2 is phylogenetically clustered with the RGD-binding receptor, and it expectedly shows the binding activity to the RGD peptide by SPR analysis.…”

“…Although information regarding the evolutionary and functional relationships among invertebrate integrins is not clear, and limited by the few genome-sequenced species, more and more integrin family members are identified in non-model invertebrates, such as in the Mollusca, which is the biggest phylum in the marine animal kingdom with the most species. The molecular aspects of some integrins in mollusks such as Lymnaea stagnalis ( 39 ), Mytilus trossulus ( 40 , 41 ), B. glabrata ( 23 , 42 ), Mytilus galloprovincialis ( 43 ), and C. gigas ( 25 , 44 – 46 ) have been described, but the complete access to information regarding any of the mollusk species remains unavailable. The Pacific oyster ( C. gigas ), a sessile filter feeder exposed to a wide range of biotic and abiotic stresses, is an attractive model to study the diversity of immune receptor families and the mediation of associated immune responses in invertebrates ( 47 , 48 ).…”

First description of a widespread Mytilus trossulus-derived bivalve transmissible cancer lineage in M. trossulus itself

“…www.nature.com/scientificreports/ Hemocytology and immunochemistry. All procedures for cell fixation and staining were described in detail in a previous study 41 . In brief, the cells were stained with TRITC-labeled phalloidin (Molecular Probes) and DAPI (Vector Laboratories) for visualization of actin cytoskeleton and nucleus.…”

Freezing tolerance of sea urchin embryonic cells: Differentiation commitment and cytoskeletal disturbances in culture

“…These and other findings [36,46,47,54] have provided enough successful protocols for cryopreservation of marine invertebrate cells and larvae that will allow researchers to work with embryonic cells from these organisms throughout the year. The development of this field not only has environmental significance, but also offers great potential for marine biotechnology, as this field may clarify the cryoresistance mechanisms of marine organisms at ultra-low temperatures.…”