Table Of ContentM M B ™
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651
wwwwwwwwwwwwww
Sphingosine-1-Phosphate
Methods and Protocols
Edited by
Alice Pébay
University of Melbourne, Parkville, Victoria, Australia
Kursad Turksen
Sprott Centre for Stem Cell Research, Regenerative Medicine Program,
Ottawa Hospital Research Institute, Ottawa, Canada
Editors
Alice Pébay Kursad Turksen
University of Melbourne Sprott Centre for Stem Cell Research
Parkville, Victoria
Regenerative Medicine Program
Australia
Ottawa Hospital Research Institute
Ottawa, Canada
ISSN 1064-3745 ISSN 1940-6029 (electronic)
ISBN 978-1-61779-799-6 ISBN 978-1-61779-800-9 (eBook)
DOI 10.1007 / 978-1-61779-800-9
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2012935580
© Springer Science+Business Media, LLC 2012
This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is
concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction
on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation,
computer software, or by similar or dissimilar methodology now known or hereafter developed. Exempted from this
legal reservation are brief excerpts in connection with reviews or scholarly analysis or material supplied specifically for
the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work.
Duplication of this publication or parts thereof is permitted only under the provisions of the Copyright Law of the
Publisher’s location, in its current version, and permission for use must always be obtained from Springer. Permissions
for use may be obtained through RightsLink at the Copyright Clearance Center. Violations are liable to prosecution
under the respective Copyright Law.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not
imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and
regulations and therefore free for general use.
While the advice and information in this book are believed to be true and accurate at the date of publication, neither
the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be
made. The publisher makes no warranty, express or implied, with respect to the material contained herein.
Printed on acid-free paper
Humana Press is a brand of Springer
Springer is part of Springer Science+Business Media (www.springer.com)
Preface
This volume of MiMB aims at describing techniques used by worldwide experts of the S1P
fi eld in an array of cell types and with various physiological relevance, hence showcasing the
important effects of S1P in development and in physiopathology.
We would like to thank all the authors of this volume who have shared their expertise
and protocols.
We also wish to thank Dr. John Walker for his support during the process of compiling
this book.
Our thanks also go to Patrick Marton for his encouragements and help.
Finally, a special thank you goes to David Casey for his tremendous help in the produc-
tion of the volume.
Parkville, Victoria, Australia Alice Pébay
Ottawa, Canada Kursad Turksen
v
wwwwwwwwwwwwww
Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 Biochemical Methods for Quantifying Sphingolipids: Ceramide, Sphingosine,
Sphingosine Kinase-1 Activity, and Sphingosine-1-Phosphate . . . . . . . . . . . . . . 1
Leyre Brizuela and Olivier Cuvillier
2 Isoform-Selective Assays for Sphingosine Kinase Activity. . . . . . . . . . . . . . . . . . 21
Melissa R. Pitman, Duyen H. Pham, and Stuart M. Pitson
3 Quantification of Sphingosine-1-Phosphate and Related Sphingolipids
by Liquid Chromatography Coupled to Tandem Mass Spectrometry. . . . . . . . . 33
Constantin Bode and Markus H. Gräler
4 A Cleanup Method for Mass Spectrometry of Sphingosine-1-Phosphate
in Blood and Solid Tissues Using a Phosphate Capture Molecule . . . . . . . . . . . 45
Jun-ichi Morishige, Tamotsu Tanaka, and Kiyoshi Satouchi
5 Immunohistochemical Detection of Sphingosine-1-Phosphate
and Sphingosine Kinase-1 in Human Tissue Samples. . . . . . . . . . . . . . . . . . . . . 55
Barbara Visentin, Gary Reynolds, and Roger Sabbadini
6 Assessment of Sphingosine-1-Phosphate Activity in Biological Samples
by Receptor Internalization and Adherens Junction Formation . . . . . . . . . . . . . 69
Hideru Obinata and Timothy Hla
7 High-Throughput Assays to Measure Intracellular Ca2+ Mobilization
in Cells that Express Recombinant S1P Receptor Subtypes . . . . . . . . . . . . . . . . 77
William J. Valentine and Gabor Tigyi
8 Quantifying Sphingosine-1-Phosphate-Dependent Activation
of the RhoGTPases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
Matt D. Medlin, Joan M. Taylor, and Christopher P. Mack
9 Optimized Protocols to Analyze Sphingosine-1-Phosphate Signal
Transduction Pathways During Acrosomal Exocytosis in Human Sperm . . . . . . 99
Silvia A. Belmonte and Laila Suhaiman
10 Use of Intravital Microscopy and In Vitro Chemotaxis Assays
to Study the Roles of Sphingosine-1-Phosphate in Bone Homeostasis. . . . . . . . 129
Taeko Ishii, Shunsuke Kawamura, Issei Nishiyama, Junichi Kikuta,
and Masaru Ishii
11 Assessment of Sphingosine-1-Phosphate Receptor Expression
and Associated Intracellular Signaling Cascades in Primary Cells
of the Human Central Nervous System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
Veronique E. Miron, Bryce A. Durafourt, Jack P. Antel,
and Timothy E. Kennedy
vii
viii Contents
12 Sphingosine-1-Phosphate Signaling in Skeletal Muscle Cells . . . . . . . . . . . . . . . 155
Elisabetta Meacci, Francesca Bini, and Chiara Battistini
13 Maintenance of Human Embryonic Stem Cells by Sphingosine-1-Phosphate
and Platelet-Derived Growth Factor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Raymond C.B. Wong, Martin F. Pera, and Alice Pébay
14 Ceramide and Sphingosine-1-Phosphate Signaling in Embryonic Stem Cell
Differentiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177
Erhard Bieberich
15 Sphingosine-1-Phosphate Signaling in Neural Progenitors. . . . . . . . . . . . . . . . . 193
Phillip Callihan and Shelley B. Hooks
16 Utilizing Sphingosine-1-Phosphate to Stimulate Sprouting Angiogenesis. . . . . . 201
Shih-Chi Su and Kayla J. Bayless
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 215
Contributors
JACK P. ANTEL (cid:129) Neuroimmunology Unit, The Montreal Neurological Institute,
McGill University, Montreal, QC, Canada
CHIARA BATTISTINI (cid:129) Department of Biochemical Sciences, University of Florence,
Florence, Italy
KAYLA J. BAYLESS (cid:129) Department of Molecular & Cellular Medicine, Texas A&M Health
Science Center, College Station, TX , USA
SILVIA A. BELMONTE (cid:129) Laboratorio de Biología Celular y Molecular, Instituto de
Histología y Embriología (IHEM-CONICET), Facultad de Ciencias Médicas,
Universidad Nacional de Cuyo, Mendoza, Argentina
ERHARD BIEBERICH (cid:129) Program in Developmental Neurobiology, Institute of Molecular
Medicine and Genetics, Medical College of Georgia, Georgia Health Sciences University,
Augusta, GA , USA
FRANCESCA BINI (cid:129) Department of Biochemical Sciences, University of Florence,
Florence, Italy
CONSTANTIN BODE (cid:129) Molecular Cancer Research Centre, Charité University Medical
School , Berlin , Germany
LEYRE BRIZUELA (cid:129) Institut de Pharmacologie et de Biologie Structurale, CNRS,
Toulouse , France ; UPS, IPBS, Université de Toulouse, Toulouse , France
PHILLIP CALLIHAN (cid:129) Department of Pharmaceutical and Biomedical Sciences,
University of Georgia, Athens , GA , USA
OLIVIER CUVILLIER (cid:129) Institut de Pharmacologie et de Biologie Structurale, CNRS,
Toulouse , France ; UPS, IPBS, Université de Toulouse, Toulouse , France
BRYCE A. D URAFOURT (cid:129) Neuroimmunology Unit, The Montreal Neurological Institute,
McGill University, Montreal , QC , Canada
MARKUS H. GRÄLER (cid:129) Molecular Cancer Research Centre, Charité University Medical
School , Berlin , Germany
TIMOTHY HLA (cid:129) Center for Vascular Biology, Department of Pathology and Laboratory
Medicine, Weill Cornell Medical College, Cornell University, New York , NY , USA
SHELLEY B. HOOKS (cid:129) Department of Pharmaceutical and Biomedical Sciences,
University of Georgia, Athens , GA , USA
MASARU ISHII (cid:129) Laboratory of Cellular Dynamics, WPI-Immunology Frontier Research
Center, Osaka University, Suita , Japan
TAEKO ISHII (cid:129) Laboratory of Cellular Dynamics, WPI-Immunology Frontier Research
Center, Osaka University, Suita , Japan
SHUNSUKE KAWAMURA (cid:129) Laboratory of Cellular Dynamics, WPI-Immunology Frontier Re-
search Center, Osaka University , Suita, Japan
TIMOTHY E. KENNEDY (cid:129) Centre for Neuronal Survival, The Montreal Neurological Insti-
tute, McGill University, Montreal , Canada
JUNICHI KIKUTA (cid:129) Laboratory of Cellular Dynamics, WPI-Immunology Frontier
Research Center, Osaka University , Suita, Japan
ix
x Contributors
CHRISTOPHER P. MACK (cid:129) Laboratory Medicine, Department of Pathology,
University of North Carolina, Chapel Hill, NC , USA
ELISABETTA MEACCI (cid:129) Department of Biochemical Sciences, University of Florence,
Florence, Italy ; Interuniversity Institute of Myology (IIM), Florence, Italy
MATT D. MEDLIN (cid:129) Laboratory MedicineDepartment of Pathology and Laboratory Medi-
cine , University of North Carolina, Chapel Hill, NC , USA
VERONIQUE E. MIRON (cid:129) Centre for Multiple Sclerosis Research, Scottish Centre for
Regenerative Medicine, The University of Edinburgh, Edinburgh, UK
JUN-ICHI MORISHIGE (cid:129) Research Center for Green Science, Fukuyama University,
Fukuyama, Japan
ISSEI NISHIYAMA (cid:129) Laboratory of Cellular Dynamics, WPI-Immunology Frontier
Research Center, Osaka University , Suita, Japan
HIDERU OBINATA (cid:129) Laboratory Medicine, Department of Pathology, Center for Vascular
Biology , Weill Cornell Medical College, Cornell University, New York , NY , USA
ALICE PÉBAY (cid:129) Department of Pharmacology, Centre for Neuroscience, The University
of Melbourne, Parkville, Victoria , Australia
MARTIN F. PERA (cid:129) Eli and Edythe Broad Center for Regenerative Medicine
and Stem Cell Research, University of Southern California, Los Angeles, CA , USA ; Cen-
tre for Neuroscience, The University of Melbourne , Parkville, VIC , Australia
DUYEN H. PHAM (cid:129) Molecular Signalling Laboratory , Centre for Cancer Biology,
SA Pathology , Adelaide, SA , Australia
MELISSA R. PITMAN (cid:129) Molecular Signalling Laboratory , Centre for Cancer Biology,
SA Pathology , Adelaide, SA , Australia
STUART M. PITSON (cid:129) Molecular Signalling Laboratory , Centre for Cancer Biology,
SA Pathology , Adelaide, SA , Australia ; School of Molecular and Biomedical Science,
University of Adelaide, Adelaide, SA , Australia
GARY REYNOLDS (cid:129) Centre for Liver Research, NIHR Biomedical Research Unit,
University of Birmingham & Queen Elizabeth Hospital Birmingham NHS Trust,
Birmingham, UK
ROGER SABBADINI (cid:129) Lpath Incorporated, San Diego, CA , USA
KIYOSHI SATOUCHI (cid:129) Department of Nutrition and Life Science, Fukuyama University,
Fukuyama, Japan
SHIH-CHI SU (cid:129) Department of Molecular & Cellular Medicine, Texas A&M Health
Science Center, College Station, TX , USA
LAILA SUHAIMAN (cid:129) Laboratorio de Biología Celular y Molecular, Instituto de Histología y
Embriología (IHEM-CONICET), Facultad de Ciencias Médicas, Universidad Nacional
de Cuyo, Mendoza, Argentina
TAMOTSU TANAKA (cid:129) Department of Applied Biological Science, Fukuyama University,
Fukuyama, Japan ; Institute of Health Biosciences, University of Tokushima
Graduate School, Tokushima , Japan
JOAN M. TAYLOR (cid:129) Department of Pathology and Laboratory Medicine,
University of North Carolina, Chapel Hill, NC , USA
GABOR TIGYI (cid:129) Department of Physiology, University of Tennessee Health Science Center,
Memphis , TN , USA
