Description
Studies incorporating cell–cell interactions remain an exciting and ever-evolving discipline within the broader areas of biology. Although recent advances in genetics have provided much insight into gene- and disease-specific mechanisms, such models are often limited by tissue/organ complexity. The use of directed cell models to study the manner by which two or more cells interact has revolutionized our understanding of protein–protein interactions in a complex milieu and have provided an invaluable platform for the identification of new molecules.
The aim of Cell–Cell Interactions: Methods and Protocols is to provide a collection of diverse protocols, incorporating methods-based approaches both in vitro and in vivo. The authors of the various chapters are highly skilled experts in the development and utilization of cell–cell interaction assays. Their extensive expertise provides a detailed, step-by-step map of a reproducible protocol. Importantly, these protocols are readily adaptable for nearly any cell type and any organ system, and for this reason, I have tried to compile a diverse set of chapters incorporating numerous individual types of cells. It is my hope that this book will be both insightful to the beginner and inspiring to the experienced.
Sean P. Colgan
Contents
Preface ………………………………………………………………………………………………..v
Contributors ……………………………………………………………………………………….. ix
1 Blood–Brain Barrier and Cell–Cell Interactions: Methods for Establishing In Vitro Models of the Blood–Brain Barrier and Transport Measurements
Michael Aschner, Vanessa A. Fitsanakis, Ana Paula Marreilha dos Santos, Luisa Olivi, and Joseph P. Bressler …. 1
2 Interactions of Intestinal Epithelial Cells With Bacteria and Immune Cells: Methods to Characterize Microflora and Functional Consequences
Geraldine Canny, Alexander Swidsinski, and Beth A. McCormick …. 17
3 The Role of Junctional Adhesion Molecules in Interactions Between Vascular Cells
Triantafyllos Chavakis and Valeria Orlova ………………………………….. 37
4 Cell–Cell Interactions on Solid Matrices
Nancy A. Louis, Dionne Daniels, and Sean P. Colgan ………………….. 51
5 Cell–Cell Interactions in the Kidney: Inducible Expression of Mutant G Protein α-Subunits in Madin–Darby Canine Kidney Cells for Studies of Epithelial Cell Junction Structure and Function
Ernesto Sabath and Bradley M. Denker ………………………………………. 61
6 Nucleotide Metabolism and Cell–Cell Interactions
Holger K. Eltzschig, Thomas Weissmüller, Alice Mager, and Tobias Eckle… 73
7 Analysis of Mammalian Sperm–Egg Membrane Interactions During In Vitro Fertilization
Genevieve B. Wortzman, Allison J. Gardner, and Janice P. Evans …. 89
8 Collagen Gel Contraction Assay
Peter Ngo, Punitha Ramalingam, Jonathan A. Phillips, and Glenn T. Furuta….103
9 Methods to Assess Tissue Permeability
Juan C. Ibla and Joseph Khoury ……………………………………………….. 111
10 Bacterial–Bacterial Cell Interactions in Biofilms: Detection of Polysaccharide Intercellular Adhesins by Blotting and Confocal Microscopy
Kimberly K. Jefferson and Nuno Cerca……………………………………… 119
11 A Biochemical Method for Tracking Cholera Toxin Transport From Plasma Membrane to Golgi and Endoplasmic Reticulum
Heidi E. De Luca and Wayne I. Lencer ……………………………………… 127
12 Isolation and Culture of Murine Heart and Lung Endothelial Cells for In Vitro Model Systems
Yaw-Chyn Lim and Francis W. Luscinskas…………………………………. 141
13 Expression Cloning of Signaling Proteins Regulated by Cell Adhesion
Michelle L. Matter and Joe W. Ramos ………………………………………. 155
14 Assays for the Calcium Sensitivity of Desmosomes
Anita J. Merritt, Anthea Scothern, and Tanusree Bhattacharyya ….. 167
15 Tight Junctions and Cell–Cell Interactions
Markus Utech, Matthias Brüwer, and Asma Nusrat ……………………. 185
16 Simultaneous Immunofluorescence Detection of Cadherin and β-Catenin in Mouse Tissue Sections
Nobuko Obara ………………………………………………………………………. 197
17 In Vitro Neutrophil Transepithelial Migration
Winston Y. Lee, Alex C. Chin, Susan Voss, and Charles A. Parkos ….. 205
18 The Blot Rolling Assay: A Method for Identifying Adhesion Molecules Mediating Binding Under Shear Conditions
Robert Sackstein and Robert Fuhlbrigge …………………………………… 217
19 Cell–Cell Interaction in the Transcellular Biosynthesis of Novel ω-3-Derived Lipid Mediators
Nan Chiang and Charles N. Serhan ………………………………………….. 227
20 Isolation and Analysis of Lipid Rafts in Cell–Cell Interactions
Aimee Landry and Ramnik Xavier ……………………………………………. 251
Index ……………………………………………………………………………………………… 283