scholarly journals Understanding dynamic changes in live cell adhesion with neutron reflectometry

2014 ◽  
Vol 28 (30) ◽  
pp. 1430015 ◽  
Author(s):  
Ann Junghans ◽  
Mary Jo Waltman ◽  
Hillary L. Smith ◽  
Luka Pocivavsek ◽  
Noureddine Zebda ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Mouse Fibroblast ◽  
Neutron Reflectometry ◽  
Live Cells ◽  
Tumor Invasiveness ◽  
Endothelial Monolayers ◽  
Key Factor ◽  
Living Tissues ◽  
Quartz Substrates ◽  
Cellular Layer

Neutron reflectometry (NR) was used to examine various live cells' adhesion to quartz substrates under different environmental conditions, including flow stress. To the best of our knowledge, these measurements represent the first successful visualization and quantization of the interface between live cells and a substrate with sub-nanometer resolution. In our first experiments, we examined live mouse fibroblast cells as opposed to past experiments using supported lipids, proteins, or peptide layers with no associated cells. We continued the NR studies of cell adhesion by investigating endothelial monolayers and glioblastoma cells under dynamic flow conditions. We demonstrated that neutron reflectometry is a powerful tool to study the strength of cellular layer adhesion in living tissues, which is a key factor in understanding the physiology of cell interactions and conditions leading to abnormal or disease circumstances. Continuative measurements, such as investigating changes in tumor cell — surface contact of various glioblastomas, could impact advancements in tumor treatments. In principle, this can help us to identify changes that correlate with tumor invasiveness. Pursuit of these studies can have significant medical impact on the understanding of complex biological problems and their effective treatment, e.g. for the development of targeted anti-invasive therapies.

scholarly journals Mouse Fibroblast Cell Adhesion Studied by Neutron Reflectometry

2010 ◽  
Vol 98 (5) ◽  
pp. 793-799 ◽  
Author(s):  
Hillary L. Smith ◽  
Joseph Hickey ◽  
Michael S. Jablin ◽  
Antoinette Trujillo ◽  
James P. Freyer ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Neutron Reflectometry ◽  
Mouse Fibroblast Cell

scholarly journals Tuning endothelial monolayer adhesion: a neutron reflectivity study

2014 ◽  
Vol 306 (1) ◽  
pp. L1-L9 ◽  
Author(s):  
Luka Pocivavsek ◽  
Ann Junghans ◽  
Noureddine Zebda ◽  
Konstantin Birukov ◽  
Jaroslaw Majewski
Keyword(s):  
Cardiovascular System ◽  
Separation Distance ◽  
Interfacial Structure ◽  
Solid Boundary ◽  
Neutron Reflectivity ◽  
Flow Conditions ◽  
Endothelial Monolayers ◽  
Biophysical Study ◽  
Living Tissues ◽  
Cellular Layer

Endothelial cells, master gatekeepers of the cardiovascular system, line its inner boundary from the heart to distant capillaries constantly exposed to blood flow. Interendothelial signaling and the monolayers adhesion to the underlying collagen-rich basal lamina are key in physiology and disease. Using neutron scattering, we report the first ever interfacial structure of endothelial monolayers under dynamic flow conditions mimicking the cardiovascular system. Endothelial adhesion (defined as the separation distance ℓ between the basal cell membrane and solid boundary) is explained using developed interfacial potentials and intramembrane segregation of specific adhesion proteins. Our method provides a powerful tool for the biophysical study of cellular layer adhesion strength in living tissues.

scholarly journals DNA-Coated AFM Cantilevers for the Investigation of Cell Adhesion and the Patterning of Live Cells

2008 ◽  
Vol 120 (44) ◽  
pp. 8601-8605 ◽  
Author(s):  
Sonny C. Hsiao ◽  
Ailey K. Crow ◽  
Wilbur A. Lam ◽  
Carolyn R. Bertozzi ◽  
Daniel A. Fletcher ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Live Cells

Dual-color imaging of cytosolic and mitochondrial zinc ions in live tissues with two-photon fluorescent probes

2014 ◽  
Vol 12 (21) ◽  
pp. 3406-3412 ◽  
Author(s):  
Kailash Rathore ◽  
Chang Su Lim ◽  
Young Lee ◽  
Bong Rae Cho
Keyword(s):  
Fluorescent Probes ◽  
Live Cells ◽  
Zinc Ions ◽  
Color Imaging ◽  
Two Photon ◽  
Dual Color ◽  
Living Tissues

We have developed TP probes for [Zn2+]cyto and [Zn2+]mito, which emit TPEF at widely-separated wavelength regions. The new probes can simultaneously detect [Zn2+]cyto and [Zn2+]mito in live cells, as well as in living tissues by dual-color TPM imaging.

scholarly journals Selenoprotein T deficiency alters cell adhesion and elevates selenoprotein W expression in murine fibroblast cells

2009 ◽  
Vol 87 (6) ◽  
pp. 953-961 ◽  
Author(s):  
Aniruddha Sengupta ◽  
Bradley A. Carlson ◽  
Vyacheslav M. Labunskyy ◽  
Vadim N. Gladyshev ◽  
Dolph L. Hatfield
Keyword(s):  
Cell Adhesion ◽  
Redox Regulation ◽  
Cell Structure ◽  
Mouse Fibroblast ◽  
Fibroblast Cells ◽  
Selenoprotein W ◽  
Expression Of Genes ◽  
Selenoprotein T ◽  
Murine Fibroblast

Mammalian selenoproteins have diverse functions, cellular locations, and evolutionary histories, but all use the amino acid selenocysteine (Sec), often present in the enzyme’s active site. Only about half of mammalian selenoproteins have been functionally characterized, with most being oxidoreductases. The cellular role of selenoprotein T (SelT), manifesting a CxxU motif in a thioredoxin-like fold and localized to Golgi and the endoplasmic reticulum, is not known. To examine its biological function, we knocked down SelT expression in mouse fibroblast cells and found that SelT deficiency alters cell adhesion and enhances the expression of several oxidoreductase genes, while decreasing the expression of genes involved in cell structure organization, suggesting the involvement of SelT in redox regulation and cell anchorage. Furthermore, we found that the loss of SelT elevates expression of another selenoprotein, selenoprotein W (SepW1). SelT and SepW1 belong to the same protein family, suggesting that SepW1 may functionally compensate for SelT.

scholarly journals L-selectin regulates human neutrophil transendothelial migration

2021 ◽  
pp. jcs.250340
Author(s):  
Izajur Rahman ◽  
Aida Collado Sánchez ◽  
Jessica Davies ◽  
Karolina Rzeniewicz ◽  
Sarah Abukscem ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Direct Evidence ◽  
Transendothelial Migration ◽  
Ectodomain Shedding ◽  
Endothelial Monolayers ◽  
Tnf Α ◽  
A Cell

The migration of circulating neutrophils towards damage/infected tissue is absolutely critical to the inflammatory response. L-selectin is a cell adhesion molecule abundantly expressed on circulating neutrophils. For over two decades, neutrophil L-selectin has been assigned the exclusive role of supporting tethering and rolling – the initial stages of the multi-step adhesion cascade. Here, we provide direct evidence for L-selectin contributing to neutrophil transendothelial migration (TEM). We show that L-selectin co-clusters with PECAM-1 - a well-characterised cell adhesion molecule involved in regulating neutrophil TEM. This co-clustering behaviour occurs specifically during TEM, which serves to augment ectodomain shedding of L-selectin and expedite the time taken for TEM (TTT) to complete. Blocking PECAM-1 signalling (through mutation of its cytoplasmic tail), PECAM-1-dependent adhesion, or L-selectin shedding, led to a significant delay in the TTT. Finally, we show that co-clustering of L-selectin with PECAM-1 occurs specifically across TNF-α- but not IL-1β-activated endothelial monolayers – implying unique adhesion interactomes forming in a cytokine-specific manner. To our knowledge, this is the first report to implicate a non-canonical role for L-selectin in regulating neutrophil TEM.

scholarly journals DNA-Coated AFM Cantilevers for the Investigation of Cell Adhesion and the Patterning of Live Cells

2008 ◽  
Vol 47 (44) ◽  
pp. 8473-8477 ◽  
Author(s):  
Sonny C. Hsiao ◽  
Ailey K. Crow ◽  
Wilbur A. Lam ◽  
Carolyn R. Bertozzi ◽  
Daniel A. Fletcher ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Live Cells
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