The involvement of adherens junction components in myofibrillogenesis in cultured cardiac myocytes

Development ◽  
1992 ◽  
Vol 114 (1) ◽  
pp. 173-183 ◽  
Author(s):  
E.J. Goncharova ◽  
Z. Kam ◽  
B. Geiger
Keyword(s):  
Alternative Pathway ◽  
Three Dimensional ◽  
Adherens Junction ◽  
Intercellular Junctions ◽  
Inhibitory Effect ◽  
Newborn Rats ◽  
Cell Surfaces ◽  
Cell Matrix ◽  
Adhesive Interactions ◽  
Lateral Cell

The distribution of adherens junction (AJ) components was investigated in cultured heart myocytes. These cells, derived from either newborn rats or chick embryos, develop elaborate arrays of myofibrils which become extensive and laterally aligned following several days in culture. The Z-disks in these cells, visualized by immunolabeling with antibodies to muscle-specific alpha-actinin, exhibit a characteristic periodicity of about 2 microns and are in register with those of neighboring myofibrils throughout the sarcoplasm. Vinculin, in these cells, associates with intercellular AJ and cell-matrix adhesions. In addition, this protein is detected in periodic bands located along the lateral cell membranes corresponding to “costamers” previously described by Pardo, J.V., Siliciano, J.D. and Craig, S.W. (Proc. Natn. Acad. Sci. USA, 80, 1008). Similarly, N-cadherin, which is predominantly associated with intercellular junctions, is also detected in periodic striations located mainly on the dorsal and lateral cell surfaces. Using computer-aided three-dimensional microscopy confirmed that these vinculin- and N-cadherin-containing structures are located in extrajunctional sites, apparently associated with Z-disks of peripheral myofibrils. Based on these findings an alternative pathway is proposed for the assembly of vinculin and N-cadherin, which is not triggered by adhesive interactions with extracellular surfaces but rather by interactions at the membrane-cytoplasm interphase with the periphery of the pre-assembled myofibrils. Moreover, we present evidence that antibodies to N-cadherin, which are capable of blocking AJ formation in culture, have an inhibitory effect also on the development and alignment of myofibrils. We discuss the functional significance of the “costameric” organization of vinculin and N-cadherin and consider its involvement both in the lateral alignment of neighboring muscle cells and in the stabilization of developing myofibrils.

Three-dimensional organization and functional relationships of endocytotic compartments in pig intestinal epithelial cells

1992 ◽  
Vol 50 (1) ◽  
pp. 576-577
Author(s):  
Julian P. Heath ◽  
Buford L. Nichols ◽  
László G. Kömüves
Keyword(s):  
Electron Microscopy ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Three Dimensional ◽  
Intestinal Epithelial ◽  
Cell Surfaces ◽  
Basal Surface ◽  
Functional Relationships

The newborn pig intestine is adapted for the rapid and efficient absorption of nutrients from colostrum. In enterocytes, colostral proteins are taken up into an apical endocytotic complex of channels that transports them to target organelles or to the basal surface for release into the circulation. The apical endocytotic complex of tubules and vesicles clearly is a major intersection in the routes taken by vesicles trafficking to and from the Golgi, lysosomes, and the apical and basolateral cell surfaces.Jejunal tissues were taken from piglets suckled for up to 6 hours and prepared for electron microscopy and immunocytochemistry as previously described.

Application of stereological analysis of cell volume to isolated myocytes in culture with and without adrenergic innervation

1990 ◽  
Vol 48 (3) ◽  
pp. 416-417
Author(s):  
Jane K. Rosenthal ◽  
Dianne L. Atkins ◽  
William J. Marvin ◽  
Penny A. Krumm
Keyword(s):  
Cardiac Myocytes ◽  
Structural Changes ◽  
Three Dimensional ◽  
Adrenergic Innervation ◽  
Culture Cell ◽  
Serial Sections ◽  
Newborn Rats ◽  
Primary Culture Cell ◽  
Biological Studies ◽  
Cell Volumes

To comprehend structural changes in cardiac myocytes accompanying adrenergic innervation, it is essential that a three dimensional analysis be performed. To date, biological studies which utilize stereological methods have been limited to cells in tissue and in organs. Our laboratory has utilized current stereological techniques for measuring absolute volumes of individual myocytes in primary culture. Cell volumes are calculated for two distinct groups of cells at 96 hours in culture: isolated myocytes and myocytes innervated with adrenergic neurons (Figure 1).Cardiac myocytes are cultured from the ventricular apices of newborn rats. Cells are plated directly onto tissue culture dishes with or without preplated explants from the paravertebral thoracolumbar sympathetic chain. On day four cultures are photographed and marked for one-to-one cell location. Following conventional fixation and embeddment in eponate-12, the cells are relocated and mounted for microtomy. The cells are completely sectioned at 120nm in their parallel orientation to the surface of the dish (Figure 2). Serial sections are collected on formvar coated slotted grids and are recorded in sequence.

scholarly journals A computational framework for modeling cell–matrix interactions in soft biological tissues

2021 ◽  
Author(s):  
Jonas F. Eichinger ◽  
Maximilian J. Grill ◽  
Iman Davoodi Kermani ◽  
Roland C. Aydin ◽  
Wolfgang A. Wall ◽  
...  
Keyword(s):  
Soft Tissues ◽  
Three Dimensional ◽  
Biological Tissues ◽  
Computational Framework ◽  
Cell Matrix ◽  
Physiological Processes ◽  
Matrix Interactions ◽  
Mechanical Homeostasis ◽  
Cell Matrix Interactions ◽  
Short Time

AbstractLiving soft tissues appear to promote the development and maintenance of a preferred mechanical state within a defined tolerance around a so-called set point. This phenomenon is often referred to as mechanical homeostasis. In contradiction to the prominent role of mechanical homeostasis in various (patho)physiological processes, its underlying micromechanical mechanisms acting on the level of individual cells and fibers remain poorly understood, especially how these mechanisms on the microscale lead to what we macroscopically call mechanical homeostasis. Here, we present a novel computational framework based on the finite element method that is constructed bottom up, that is, it models key mechanobiological mechanisms such as actin cytoskeleton contraction and molecular clutch behavior of individual cells interacting with a reconstructed three-dimensional extracellular fiber matrix. The framework reproduces many experimental observations regarding mechanical homeostasis on short time scales (hours), in which the deposition and degradation of extracellular matrix can largely be neglected. This model can serve as a systematic tool for future in silico studies of the origin of the numerous still unexplained experimental observations about mechanical homeostasis.

Attractive forces slow contact formation between deformable bodies underwater

2021 ◽  
Vol 118 (41) ◽  
pp. e2104975118
Author(s):  
Mengyue Sun ◽  
Nityanshu Kumar ◽  
Ali Dhinojwala ◽  
Hunter King
Keyword(s):  
Three Dimensional ◽  
Adhesive Contact ◽  
Final State ◽  
Underwater Adhesion ◽  
Frustrated Total Internal Reflection ◽  
Solid Deformation ◽  
Hydrophilic Surfaces ◽  
Deformable Bodies ◽  
Adhesive Interactions

Thermodynamics tells us to expect underwater contact between two hydrophobic surfaces to result in stronger adhesion compared to two hydrophilic surfaces. However, the presence of water changes not only energetics but also the dynamic process of reaching a final state, which couples solid deformation and liquid evacuation. These dynamics can create challenges for achieving strong underwater adhesion/friction, which affects diverse fields including soft robotics, biolocomotion, and tire traction. Closer investigation, requiring sufficiently precise resolution of film evacuation while simultaneously controlling surface wettability, has been lacking. We perform high-resolution in situ frustrated total internal reflection imaging to track underwater contact evolution between soft-elastic hemispheres of varying stiffness and smooth–hard surfaces of varying wettability. Surprisingly, we find the exponential rate of water evacuation from hydrophobic–hydrophobic (adhesive) contact is three orders of magnitude lower than that from hydrophobic–hydrophilic (nonadhesive) contact. The trend of decreasing rate with decreasing wettability of glass sharply changes about a point where thermodynamic adhesion crosses zero, suggesting a transition in mode of evacuation, which is illuminated by three-dimensional spatiotemporal height maps. Adhesive contact is characterized by the early localization of sealed puddles, whereas nonadhesive contact remains smooth, with film-wise evacuation from one central puddle. Measurements with a human thumb and alternatively hydrophobic/hydrophilic glass surface demonstrate practical consequences of the same dynamics: adhesive interactions cause instability in valleys and lead to a state of more trapped water and less intimate solid–solid contact. These findings offer interpretation of patterned texture seen in underwater biolocomotive adaptations as well as insight toward technological implementation.

scholarly journals An Analysis Based on Molecular Docking and Molecular Dynamics Simulation Study of Bromelain as Anti-SARS-CoV-2 Variants

2021 ◽  
Vol 12 ◽  
Author(s):  
Trina Ekawati Tallei ◽  
Fatimawali ◽  
Afriza Yelnetty ◽  
Rinaldi Idroes ◽  
Diah Kusumawaty ◽  
...  
Keyword(s):  
Molecular Dynamics ◽  
Molecular Docking ◽  
Molecular Dynamics Simulation ◽  
Three Dimensional ◽  
Md Simulations ◽  
Inhibitory Effect ◽  
Dynamics Simulation ◽  
Novel Coronavirus

The rapid spread of a novel coronavirus known as SARS-CoV-2 has compelled the entire world to seek ways to weaken this virus, prevent its spread and also eliminate it. However, no drug has been approved to treat COVID-19. Furthermore, the receptor-binding domain (RBD) on this viral spike protein, as well as several other important parts of this virus, have recently undergone mutations, resulting in new virus variants. While no treatment is currently available, a naturally derived molecule with known antiviral properties could be used as a potential treatment. Bromelain is an enzyme found in the fruit and stem of pineapples. This substance has been shown to have a broad antiviral activity. In this article, we analyse the ability of bromelain to counteract various variants of the SARS-CoV-2 by targeting bromelain binding on the side of this viral interaction with human angiotensin-converting enzyme 2 (hACE2) using molecular docking and molecular dynamics simulation approaches. We have succeeded in making three-dimensional configurations of various RBD variants using protein modelling. Bromelain exhibited good binding affinity toward various variants of RBDs and binds right at the binding site between RBDs and hACE2. This result is also presented in the modelling between Bromelain, RBD, and hACE2. The molecular dynamics (MD) simulations study revealed significant stability of the bromelain and RBD proteins separately up to 100 ns with an RMSD value of 2 Å. Furthermore, despite increases in RMSD and changes in Rog values of complexes, which are likely due to some destabilized interactions between bromelain and RBD proteins, two proteins in each complex remained bonded, and the site where the two proteins bind remained unchanged. This finding indicated that bromelain could have an inhibitory effect on different SARS-CoV-2 variants, paving the way for a new SARS-CoV-2 inhibitor drug. However, more in vitro and in vivo research on this potential mechanism of action is required.

scholarly journals Tuning cell migration: contractility as an integrator of intracellular signals from multiple cues

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 1819 ◽  
Author(s):  
Francois Bordeleau ◽  
Cynthia A. Reinhart-King
Keyword(s):  
Cell Migration ◽  
Growth Factors ◽  
Three Dimensional ◽  
Two Dimensional ◽  
Multiple Cues ◽  
Cell Contractility ◽  
Cell Matrix ◽  
Intracellular Signals ◽  
Physical Cues ◽  
The Matrix

There has been immense progress in our understanding of the factors driving cell migration in both two-dimensional and three-dimensional microenvironments over the years. However, it is becoming increasingly evident that even though most cells share many of the same signaling molecules, they rarely respond in the same way to migration cues. To add to the complexity, cells are generally exposed to multiple cues simultaneously, in the form of growth factors and/or physical cues from the matrix. Understanding the mechanisms that modulate the intracellular signals triggered by multiple cues remains a challenge. Here, we will focus on the molecular mechanism involved in modulating cell migration, with a specific focus on how cell contractility can mediate the crosstalk between signaling initiated at cell-matrix adhesions and growth factor receptors.

Targeting of the myosin-I myr 3 to intercellular adherens type junctions induced by dominant active Cdc42 in HeLa cells

1998 ◽  
Vol 111 (18) ◽  
pp. 2779-2788 ◽  
Author(s):  
H.E. Stoffler ◽  
U. Honnert ◽  
C.A. Bauer ◽  
D. Hofer ◽  
H. Schwarz ◽  
...  
Keyword(s):  
Hela Cells ◽  
Adherens Junction ◽  
Intercellular Junctions ◽  
Distinct Type ◽  
Beta Catenin ◽  
Tail Domain ◽  
Filamentous Actin ◽  
Myosin I ◽  
Myosin Motor Domain ◽  
Constitutively Active

Myr 3, a member of the myosin-I family from rat, is shown in this study to be localized at adherens-type intercellular junctions in epithelial and nonepithelial tissues. Formation of intercellular junctions and the accompanying recruitment of myr 3 to these junctions involves signaling by the Rho subfamily of small GTP-binding proteins. This conclusion is based on studies with HtTA-1 HeLa cells that were induced by overexpression of constitutively active Cdc42Hs to form typical adherens-type intercellular junctions enriched in cadherins (N-cadherin), beta-catenin, filamentous actin and myr 3. Recruitement of myr 3 to Cdc42-induced adherens junctions in HeLa cells was dependent on a short region of the tail domain and a functional myosin motor domain, but was independent of its myosin-I tail homology and SH3 regions. Overexpression of constitutively active Rac1 induced a distinct type of adherens junction in HeLa cells that was characterized by elaborate intercellular interdigitations enriched in N-cadherin, beta-catenin and F-actin. Myr 3 was often present, but not specifically enriched in the intercellular junctions induced by constitutively active Rac1.

Inhibitory effect of resveratrol on the growth and angiogenesis of human endometrial tissue in an In Vitro three-dimensional model of endometriosis

2020 ◽  
Vol 20 (4) ◽  
pp. 484-490
Author(s):  
Mohammad Rasool Khazaei ◽  
Zahra Rashidi ◽  
Farzaneh Chobsaz ◽  
Elham Niromand ◽  
Mozafar Khazaei
Keyword(s):  
Three Dimensional ◽  
Inhibitory Effect ◽  
Endometrial Tissue ◽  
Dimensional Model ◽  
Three Dimensional Model

scholarly journals The molecular basis of mouse sperm–zona pellucida binding: a still unresolved issue in developmental biology

Reproduction ◽  
2011 ◽  
Vol 142 (3) ◽  
pp. 377-381 ◽  
Author(s):  
Gary F Clark
Keyword(s):  
Zona Pellucida ◽  
Molecular Basis ◽  
Three Dimensional ◽  
Inhibitory Effect ◽  
Specific Model ◽  
Cell Stage ◽  
Sperm Binding ◽  
Potent Inhibitory Effect ◽  
Alternate Model ◽  
Sperm Plasma Membrane

During murine fertilization, sperm bind to the specialized extracellular matrix of the egg, known as the zona pellucida (ZP). This matrix is composed of three major glycoproteins designated ZP1, ZP2, and ZP3. Three models for sperm–ZP binding are now under consideration. The domain-specific model posits that adhesion relies primarily on interactions between N-glycans located within the C-terminal domain of ZP3 and a lectin-like egg-binding protein in the sperm plasma membrane. However, this model does not explain recent results obtained in studies with ZP2mut mice. In the supramolecular structure model, sperm bind to a three-dimensional zona matrix that depends on the cleavage status of ZP2. This paradigm does not explain the potent inhibitory effect of specific carbohydrate sequences or a C-terminal glycopeptide (gp55) derived from ZP3. Recently, O-glycans linked at Thr155 and Thr162 of ZP3 were implicated as potential ligands that mediate initial sperm–ZP binding. This novel model will be reviewed. A major challenge is to develop an alternate model for sperm–ZP binding that fits as much of the data as possible. Such a model is presented in this review. This paradigm could explain how the inability to cleave ZP2mut in ZP2mut mice could result in continued sperm binding to two-cell stage embryos without the formation of a supramolecular binding complex. These novel insights should guide future experiments that will eventually determine the molecular basis underlying gamete binding in the mouse and other eutherian mammals.

Sign in / Sign up

Export Citation Format

Share Document