Cerebral Angiogenesis: Methods and Protocols

441 Pages·2014·10.82 MB·English

by  Richard Milner (eds.)

Methods in
Molecular Biology 1135
Richard Milner Editor
Cerebral
Angiogenesis
Methods and Protocols
M M B
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hat fi eld, Hertfordshire, AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651

Cerebral Angiogenesis
Methods and Protocols
Edited by
Richard Milner
Department of Molecular and Experimental Medicine,
The Scripps Research Institute, La Jolla, CA, USA
Editor
Richard M ilner
Department of Molecular and Experimental Medicine
The Scripps Research Institute
La Jolla , CA, USA
ISSN 1064-3745 ISSN 1940-6029 (electronic)
ISBN 978-1-4939-0319-1 ISBN 978-1-4939-0320-7 (eBook)
DOI 10.1007/978-1-4939-0320-7
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Prefa ce
While blood vessels in the adult central nervous system are extremely stable under normal
conditions, it is now accepted that these vessels can be rapidly remodeled in a variety of
physiological and pathological situations, including cerebral hypoxia, cerebral ischemia,
and tumor development. All of these conditions trigger cerebral angiogenesis, i.e., the
sprouting of new vessels. In light of the critical importance of cerebral angiogenesis in pro-
moting physiological adaptation (hypoxia), recovery (cerebral ischemia), or pathogenesis
(brain tumors), it becomes a high priority to fully understand the molecular mechanisms
underlying this process. In the last 20 years, we have tremendously expanded our working
knowledge of how vascular remodeling in the brain occurs and identifi ed many of the key
cellular and molecular events underlying this process. From a clinical viewpoint, clear goals
are to promote angiogenesis following ischemic stroke or block angiogenic remodeling
during brain tumor formation. However, we are still some way off from achieving these
noble aims, and it is likely that only after more intensive research and studies, based on the
kind of principles and experimental approaches outlined in this volume, will these goals be
realistically attainable.
In light of the rapidly expanding fi eld of cerebral angiogenesis, the aim of C erebral
Angiogenesis: Methods and Protocols is to provide a selection of the key techniques that are
used in characterizing cerebral angiogenesis and to defi ne the cellular and molecular mech-
anisms underlying this important process. Following the objective of the highly successful
Methods in Molecular Biology series, the aim of this book is to provide a comprehensive step-
by- step guide for the many models and techniques for studying cerebral angiogenesis as
well as more specialized approaches for examining endothelial cell behavior, both in vitro
and in vivo. As such, Cerebral Angiogenesis: Methods and Protocols should help all research-
ers, both those new to this fi eld as well as those looking to use more specialized and sophis-
ticated techniques, to examine blood vessel growth in the brain. Cerebral Angiogenesis:
Methods and Protocols covers a wide range of protocols, which provide a useful resource for
vascular biologists, cell biologists, molecular biologists, and neuroscientists. This book is
divided into six parts. Part I provides an overview of cerebral angiogenesis occurring in dif-
ferent scenarios. Part II describes a variety of different models in which cerebral angiogen-
esis can be studied as well as includes one chapter describing cerebral arteriogenesis, the
formation of new functional arteries. Part III presents a number of methods to characterize
and quantify angiogenic events as well as several different approaches to measure changes
in cerebral blood fl ow. Part IV describes different approaches to investigate the role of spe-
cifi c candidate genes in cerebral angiogenesis as well as an important method to determine
the contribution of blood-derived hematopoietic stem cells. Part V presents several meth-
ods of therapeutically manipulating cerebral angiogenesis by gene delivery. Finally, Part VI
outlines some important approaches to examine cerebral angiogenic mechanisms in vitro,
including primary culture of brain endothelial cells from different species, assays to examine
different aspects of endothelial cell function, and cell culture systems to examine blood
vessel formation in vitro. It also includes two chapters that describe in vitro systems of the
blood–brain barrier.
v
vi Preface
I hope that all readers, both fi rst year graduate students and experienced investigators,
will fi nd Cerebral Angiogenesis: Methods and Protocols to be both timely and useful in their
research endeavours. I would like to thank all contributors for their valuable time and effort
spent in preparing these excellent chapters and for passing on the critical steps and snippets
of experimental knowledge, painstakingly acquired over many years in the laboratory, that
can make all of our lives that much easier. My sincere thanks also go to John Walker for
giving me the opportunity to put this collection together and for providing editorial assis-
tance during the preparation of this book.
La Jolla, C A, USA Richard Milner
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
PART I OVERVIEW OF CEREBRAL ANGIOGENESIS
1 Cerebral Angiogenesis During Development:
Who Is Conducting the Orchestra?. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Ina M. Wittko-Schneider, Fabian T. Schneider, and Karl H. Plate
2 Cerebral Angiogenesis: A Realistic Therapy for Ischemic Disease? . . . . . . . . . . 21
David A. Greenberg
3 Vascular Normalization in Cerebral Angiogenesis: Friend or Foe? . . . . . . . . . . 25
Jisook Lee, Andrew Baird, and Brian P. Eliceiri
4 Pericytes and Adaptive Angioplasticity: The Role of Tumor Necrosis
Factor-Like Weak Inducer of Apoptosis (TWEAK) . . . . . . . . . . . . . . . . . . . . . 35
Paula Dore-Duffy
PART II ANIMAL MODELS OF CEREBRAL ANGIOGENESIS
5 Analysis of Angiogenesis in the Developing Mouse
Central Nervous System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Nicole Ziegler, Karl H. Plate, and Stefan Liebner
6 Hypoxia-Induced Angiogenesis and Capillary Density Determination . . . . . . . 69
Constantinos P. Tsipis, Xiaoyan Sun, Kui Xu, and Joseph C. LaManna
7 The Middle Cerebral Artery Occlusion Model of Transient Focal
Cerebral Ischemia. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Fudong Liu and Louise D. McCullough
8 A Mouse Model of Chronic Cerebral Hypoperfusion Characterizing
Features of Vascular Cognitive Impairment. . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Masafumi Ihara, Akihiko Taguchi, Takakuni Maki, Kazuo Washida,
and Hidekazu Tomimoto
9 A Mouse Model of Permanent Focal Ischemia:
Distal Middle Cerebral Artery Occlusion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103
Kristian P. Doyle and Marion S. Buckwalter
10 A Method of Inducing Global Cerebral Ischemia . . . . . . . . . . . . . . . . . . . . . . 111
Gina Hadley, Michalis Papadakis, and Alastair M. Buchan
11 Induction of Cerebral Arteriogenesis in Mice . . . . . . . . . . . . . . . . . . . . . . . . . 121
André Duelsner, Nora Gatzke, Anja Bondke Persson,
and Ivo R. Buschmann
vii
viii Contents
12 Vessel Painting Technique for Visualizing
the Cerebral Vascular Architecture of the Mouse. . . . . . . . . . . . . . . . . . . . . . . 127
Shea Hughes, Oleksandr Dashkin, and Richard Anthony DeFazio
13 Examining Cerebral Angiogenesis in Response to Physical Exercise. . . . . . . . . 139
Kiersten L. Berggren, Jacob J.M. Kay, and Rodney A. Swain
PART III METHODS OF EXAMINING CEREBRAL ANGIOGENESIS
14 Histological Assessment of Angiogenesis
in the Hypoxic Central Nervous System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
Moises Freitas-Andrade, Jacqueline Slinn, Claudie Charlebois,
and Maria J. Moreno
15 Examining Vascular Remodeling in the Hypoxic Central Nervous System . . . . 177
Amin Boroujerdi, Jennifer V. Welser-Alves, and Richard Milner
16 Analysis of Cerebral Angiogenesis in Human Glioblastomas . . . . . . . . . . . . . . 187
Michel Mittelbronn, Peter Baumgarten, Patrick N. Harter,
and Karl H. Plate
17 Quantitative Cerebral Blood Flow Measurements Using MRI. . . . . . . . . . . . . 205
Eric R. Muir, Lora Talley Watts, Yash Vardhan Tiwari,
Andrew Bresnen, Qiang Shen, and Timothy Q. Duong
18 Fluorescent Angiogenesis Models Using Gelfoam®
Implanted in Transgenic Mice Expressing Fluorescent Proteins. . . . . . . . . . . . 213
Robert M. Hoffman
19 Laser Speckle Contrast Imaging to Measure Changes
in Cerebral Blood Flow. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223
Ian R. Winship
20 Laser Doppler Flowmetry to Measure Changes in Cerebral Blood Flow . . . . . 237
Brad A. Sutherland, Tamer Rabie, and Alastair M. Buchan
PART IV DETERMINING THE ROLE OF CANDIDATE GENES
IN CEREBRAL ANGIOGENESIS
21 Defining the Role of HIF and Its Downstream Mediators
in Hypoxic-Induced Cerebral Angiogenesis. . . . . . . . . . . . . . . . . . . . . . . . . . . 251
Xiaoyan Sun, Constantinos P. Tsipis, Girriso F. Benderro, Kui Xu,
and Joseph C. LaManna
22 Inducible Gene Deletion in Glial Cells to Study Angiogenesis
in the Central Nervous System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261
Hye Shin Lee and Joseph H. McCarty
23 Bone Marrow Chimera Experiments to Determine the Contribution
of Hematopoietic Stem Cells to Cerebral Angiogenesis. . . . . . . . . . . . . . . . . . 275
Marcia Regina Machein and Karl H. Plate
Contents ix
24 Novel Methods for Accurate Identification, Isolation, and Genomic
Analysis of Symptomatic Microenvironments in Atherosclerotic Arteries . . . . . 289
Mark Slevin, Maribel Baldellou, Elspeth Hill, Yvonne Alexander,
Garry McDowell, Christopher Murgatroyd, Michael Carroll,
Hans Degens, Jerzy Krupinski, Norma Rovira,
Mohammad Chowdhury, Ferdinand Serracino-I nglott,
and Lina Badimon
PART V STIMULATION OF CEREBRAL ANGIOGENESIS BY GENE DELIVERY
25 Induction of Brain Arteriovenous Malformation in the Adult Mouse. . . . . . . . 309
Wanqiu Chen, William L. Young, and Hua Su
26 Stimulation of Cerebral Angiogenesis by Gene Delivery . . . . . . . . . . . . . . . . . 317
Yaohui Tang, Yaning Li, Xiaojie Lin, Peng Miao, Yongting Wang,
and Guo-Yuan Yang
27 Investigating the Role of Perlecan Domain V
in Post-I schemic Cerebral Angiogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 331
Aileen Marcelo and Gregory Bix
PART VI METHODS TO STUDY CEREBRAL ANGIOGENESIS IN VITRO
28 Isolation and Culture of Primary Mouse Brain Endothelial Cells. . . . . . . . . . . 345
Jennifer V. Welser-Alves, Amin Boroujerdi, and Richard Milner
29 Purification of Endothelial Cells from Rat Brain . . . . . . . . . . . . . . . . . . . . . . . 357
Jinhua Luo, Xiangling Yin, Alma Sanchez, Debjani Tripathy,
Joseph Martinez, and Paula Grammas
30 Generation of Primary Cultures of Bovine Brain Endothelial Cells
and Setup of Cocultures with Rat Astrocytes. . . . . . . . . . . . . . . . . . . . . . . . . . 365
Hans C. Helms and Birger Brodin
31 Isolation and Culture of Primary Pericytes from Mouse Brain . . . . . . . . . . . . . 383
Amin Boroujerdi, Ulrich Tigges, Jennifer V. Welser-Alves,
and Richard Milner
32 Assays to Examine Endothelial Cell Migration,
Tube Formation, and Gene Expression Profiles. . . . . . . . . . . . . . . . . . . . . . . . 393
Shuzhen Guo, Josephine Lok, Yi Liu, Kazuhide Hayakawa,
Wendy Leung, Changhong Xing, Xunming Ji, and Eng H. Lo
33 A Neurovascular Blood–Brain Barrier In Vitro Model. . . . . . . . . . . . . . . . . . . 403
Christoph M. Zehendner, Robin White, Jana Hedrich,
and Heiko J. Luhmann
34 In Vitro Models of the Blood–Brain Barrier . . . . . . . . . . . . . . . . . . . . . . . . . . 415
Cathrin J. Czupalla, Stefan Liebner, and Kavi Devraj
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 439