Animal Models of Acute Neurological Injuries II: Injury and Mechanistic Assessments, Volume 2

713 Pages·2012·13.66 MB·English

by  R. Anne Stetler, Miguel A. Perez-Pinzon (auth.), Jun Chen, Xiao-Ming Xu, Zao C. Xu, John H. Zhang (eds.)

S P H
PRINGER ROTOCOLS ANDBOOKS
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Animal Models of Acute
Neurological Injuries II
Injury and Mechanistic Assessments, Volume 2
Edited by
Jun Chen
Department of Neurology and the Center for Cerebrovascular
Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
Xiao-Ming Xu
Departments of Neurological Surgery and Anatomy & Cell Biology,
Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute,
Indiana University School of Medicine, Indianapolis, IN, USA
Zao C. Xu
Department of Anatomy & Cell Biology, Indiana University School of Medicine,
Indianapolis, IN, USA
John H. Zhang
Department of Neurosurgery, Loma Linda University School of Medicine,
Loma Linda, CA, USA
Editors
Jun Chen Xiao-Ming Xu
Department of Neurology Departments of Neurological Surgery and
and the Center for Cerebrovascular Anatomy & Cell Biology
Disease Research Spinal Cord and Brain Injury Research Group
University of Pittsburgh School of Medicine Stark Neurosciences Research Institute
Pittsburgh, PA, USA Indiana University School of Medicine
Indianapolis, IN, USA
Zao C. Xu
Department of Anatomy & Cell Biology John H. Zhang
Indiana University School of Medicine Department of Neurosurgery
Indianapolis, IN, USA Loma Linda University School of Medicine
Loma Linda, CA, USA
ISSN 1949-2448 e-ISSN 1949-2456
ISBN 978-1-61779-781-1 e-ISBN 978-1-61779-782-8
DOI 10.1007/978-1-61779-782-8
Springer New York Dordrecht Heidelberg London
Library of Congress Control Number: 2012933796
© Springer Science+Business Media, LLC 2012
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Foreword
The second edition of this outstanding reference book is being published in very challenging
times. Ischemic and traumatic CNS injuries remain a huge public health problem through-
out the world. Stroke continues to be ranked as the third leading cause of death in all
developed countries and arguably the major cause of long-term disability. Within the USA
alone, estimates are currently at 1.2 million stroke incidents each year, leading to over
150,000 outright deaths. The picture for traumatic brain injury (TBI) and spinal cord
injury (SCI) is equally daunting and impacts the young and the very old of our population
disproportionately. In Europe, head-injured individuals living with signifi cant disability
number near seven million. In China, estimates indicate that over three million people each
year endure signifi cant brain injury. These facts emphasize the need for innovative approaches
to CNS injury research and a reinvigoration of our mandate to translate data from bench-
to-bedside.
Equally as challenging is the lack of safe pharmacological agents that consistently
improve clinical CNS injury outcomes. Little new pharmacology has entered clinical prac-
tice for TBI in over three decades, and even investigational therapies, such as hypothermia
remain uncertain in safety or effi cacy for heterogeneous patient populations. At this writ-
ing, the stroke research community is thoughtfully questioning the achievability of thera-
peutic “neuroprotection” in humans. The latest large-scale clinical trial failure is still fresh
in our minds, as we review summaries of the Stroke Acute Ischemic NXY-059 Treatment
Trial (SAINT II). Bench a nd clinical researchers are looking for signs of hopeful progress,
despite SAINT II’s negative results. Many an international stroke conference opens its ses-
sions with the reminder that recombinant tissue plasminogen activator (tPA) remains the
only FDA approved agent for acute stroke treatment. While tPA does not repair post-
ischemic cellular damage, its effi cacy emphasizes the oldest research concept in our arma-
mentarium, i.e., the importance of restoring perfusion. Studies of optimal recanalization
remain crucial, fueled by the extension of tPA’s therapeutic window beyond 3 h. The latter
aspect may widen tPA’s utility in acute stroke therapy in a variety of countries.
Despite this litany of challenges, basic and translational CNS injury researchers can
point to many areas of progress and changing experimental paradigms that are vigorous and
productive. Compelling directions include a focal shift from neuron-centric pharmacologi-
cal agents into (1) integrated evaluations of the neurovascular unit, (2) increased investiga-
tional intensity into neuro-regeneration and the therapeutic boundary between molecular
protection vs. recovery mechanisms, (3) a broader research stance that addresses stroke as
a “systemic disease” that impacts non-CNS organs and the immune system, and (4) creative
endovascular approaches to clot retrieval and manipulation.
Essential to all of these challenges and opportunities is the understanding, and precise
use, of animal models in CNS injury research. In this second edition, Drs. Jun Chen, Xiao-
Ming Xu, Zao Xu, and John Zhang provide a vital reference that will advance our fi eld and
educate researchers at all levels. These substantive volumes provide a manual for expert use
of animal models of cerebral ischemia, intracerebral and subarachnoid hemorrhage, vasos-
pasm, TBI, and SCI and emphasize mechanistic evaluations in all areas. An impressive array
v
vi Foreword
of internationally valued experts and leading investigators have authored chapters which are
oriented around disease processes, culminating in a concise methodological reference that
the reader will depend on as a guide to success. The volumes have been thoroughly updated
and emphasize procedures in a way that captures underlying cellular and molecular mecha-
nisms at work in injured tissue. Helpful material outlines procedural pitfalls and ways to
avoid common failures that can confound in vivo studies.
For newcomers to CNS injury, Part I through V of Volume 1 are essential as they sur-
vey the principles and technical nuances of all current methodologies appropriate for com-
prehensive animal studies. Part VI–VIII focus on hemorrhage and vasospasm, a topic of
great importance as patients with these diseases are commonly found in neuroscience criti-
cal care units around the world. Part I and II of Volume 2 contain the foundations of CNS
ischemic models and offer broad coverage of morphological, physiological, molecular, and
behavioral assessments, with new emphasis on neurogenesis and recovery parameters. Part
III focuses on neonatal injury, a highly specialized and under-studied area of ischemic
pathology that is frequently overlooked in reference compendia. Part IV and V complete
the survey of injury models with key material on the assessment of TBI and SCI, emphasiz-
ing up-to-date imaging techniques that are elemental to the fi eld.
The use of sophistical animal models remains the bedrock of CNS injury research.
There is currently no substitute for the intact animal if our goal is to evaluate microcircula-
tory networks, the cells and units these networks serve, or the integrated functional output
that is the basis of life. A nimal Models of Acute Neurological Injuries II is an invaluable aid
to junior investigators and more senior scientists who are forging new directions in their
science or perhaps engaging translational hypotheses for the fi rst time. This valuable collec-
tion of experience from the best in the fi eld offers a thoughtful opportunity to all of us
faced with challenging times.
Portland, OR, USA Patricia D. Hurn
Preface
Our fi rst book, entitled A nimal Models of Acute Neurological Injuries , published by Humana
Press/Springer (ISBN: 978-1-60327-184-4), was a great success. In that book, we con-
structed a benchwork manual for the most commonly used animal models of acute neuro-
logical injuries, including cerebral ischemia, hemorrhage, vasospasm, and traumatic brain,
and spinal cord injuries. Since its publication, the book has been well received by both clini-
cal and basic researchers due to its practicality.
Since the animal models for acute neurological injuries are in place, our next goal, i.e.,
the goal of these books, was to construct chapters on assessing these disorders from cells
and molecules to behavior and imaging. These comprehensive assessments are the key for
understanding disease mechanisms as well as developing novel therapeutic strategies to
ameliorate or even prevent damages to the nervous system.
The layout of these books is disease/disorder oriented, which encapsulates several parts,
including sections on global cerebral ischemia, focal cerebral ischemia, neonatal hypoxia–
ischemia, subarachnoid hemorrhage, cerebral vasospasm, intracerebral hemorrhage, trau-
matic brain injury, traumatic spinal cord injury, and general assessments. Each part includes
coverage of morphological, physiological, biochemical, neurobehavioral, and neuroimag-
ing assessments. In general, each begins with an introductory discussion on the availability
and selection of specifi c assessments for specifi c injury models as well as their pros and cons.
In some chapters, the assessments are divided according to levels of importance into “rec-
ommended methods” that are well established and most extensively used, or “alternative
methods” that depict less extensively used assessments that have their own utility.
These books are designed to provide both expert guidance and step-by-step proce-
dures, along with multiple photographs and/or schematic drawings on assessments of acute
neurological injuries. Throughout each chapter, the readers are aided in understanding
what, why, when, where, and how a particular assessment is used. We hope that these books
can be useful for trainees or beginners in their assessments of acute neurological injuries, for
experienced scientists from other research fi elds who are interested in either switching fi elds
or exploring new opportunities, and for established scientists within the fi eld who wish to
employ new assessments.
We are grateful to all chapter contributors for their time, effort, and dedication. It would
be impossible to publish these books without their signifi cant contributions. Finally, we
hope that the volumes will be useful for students, postdoctoral fellows, and clinical and
basic scientists who would like to study acute neurological disorders.
Pittsburg, PA, USA Jun Chen
Indianapolis, IN, USA Xiao-Ming Xu
Indianapolis, IN, USA Zao C. Xu
Loma Linda, CA, USA John H. Zhang
vii

Contents
Foreword . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiii
1 Introduction (Parts I–III). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
R. Anne Stetler and Miguel A. Perez-Pinzon
PART I GLOBAL CEREBRAL ISCHEMIA
2 Morphological Assessments of Global Cerebral Ischemia: Viable Cells. . . . . . . . . . . 7
Peiying Li
3 Morphological Assessments of Global Cerebral Ischemia: Degenerated Cells. . . . . . 19
Peiying Li
4 Morphological Assessments of Global Cerebral Ischemia: Electron Microscopy. . . . 29
Irina S. Balan, Tibor Kristian, Chunli Liu, Andrew J. Saladino,
and Bingren Hu
5 Biochemical and Molecular Biological Assessments of Global Cerebral
Ischemia: mRNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
R. Anne Stetler and Yu Gan
6 Biochemical and Molecular Biological Assessments of Global Cerebral
Ischemia: Protein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
Yu Gan and R. Anne Stetler
7 Neurobehavioral Assessments of Global Cerebral Ischemia . . . . . . . . . . . . . . . . . . . 67
Pinar Kendigelen, Dandan Sun, and Pelin Cengiz
8 Assessment of Neurogenesis in the Dentate Gyrus After Global
or Focal Ischemia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Wei Liu and Jialing Liu
PART II FOCAL CEREBRAL ISCHEMIA
9 Infarct Measurement in Focal Cerebral Ischemia: TTC Staining . . . . . . . . . . . . . . . 93
Feng Zhang and Jun Chen
10 Morphological Assessments of Focal Cerebral Ischemia: White Matter Injury. . . . . 99
Feng Zhang and Lyanne C. Schlichter
11 Blood Flow Reduction: Laser Doppler, 14C-IAP. . . . . . . . . . . . . . . . . . . . . . . . . . . 107
Feng Zhang, Steven H. Graham, and Jun Chen
12 Biochemical and Molecular Biological Assessments of Focal Cerebral
Ischemia: mRNA and MicroRNA. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
Yu Gan and R. Anne Stetler
ix
x Contents
13 Biochemical and Molecular Biological Assessments of Focal Cerebral
Ischemia: Protein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
R. Anne Stetler and Yu Gan
14 Assessments of Inflammation After Focal Cerebral Ischemia . . . . . . . . . . . . . . . . . . 139
Xiaoming Hu and Jun Chen
15 Neurobehavioral Assessments of Focal Cerebral Ischemia:
Sensorimotor Deficit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
Yejie Shi and Dandan Sun
16 Neurobehavioral Assessments of Focal Cerebral Ischemia: Cognitive Deficit. . . . . . 157
Yejie Shi and Dandan Sun
17 Assessment of Neurogenesis in Models of Focal Cerebral Ischemia. . . . . . . . . . . . . 163
Feng-Yan Sun, Xiao Sun, Ji-Jiang Wang, Qiu-Wan Zhang,
and Yong-Quan Wang
18 Assessment of Angiogenesis in Models of Focal Cerebral Ischemia . . . . . . . . . . . . . 181
Hong Shi, Yanqin Gao, and Weimin Liang
PART III NEONATAL HYPOXIA-ISCHEMIA
19 Morphological Assessments of Neonatal Hypoxia–Ischemia: In Situ Cell
Degeneration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
Changlian Zhu and Klas Blomgren
20 Morphological Assessments of Neonatal Hypoxia–Ischemia:
White Matter and Blood-Brain Barrier Injury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201
Dianer Yang and Chia-Yi Kuan
21 Biochemical and Molecular Biological Assessments of Neonatal
Hypoxia–Ischemia: Cell Signaling. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211
Changlian Zhu and Klas Blomgren
22 Biochemical and Molecular Biological Assessments of Neonatal
Hypoxia–Ischemia: Inflammation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221
Xiaoming Hu and Jun Chen
23 Neurobehavioral Assessments of Neonatal Hypoxia–Ischemia. . . . . . . . . . . . . . . . . 223
Masanori Iwai and Jun Chen
24 Assessment of Neurogenesis and White Matter Regeneration . . . . . . . . . . . . . . . . . 229
Masanori Iwai, Hiroko Tajiri, Shiro Matsumoto, Hiroshi Mitsubuchi,
and Fumio Endo
PART IV TRAUMATIC BRAIN INJURY
25 Assessments for Traumatic Brain Injury: An Introduction. . . . . . . . . . . . . . . . . . . . 247
Kathryn E. Saatman
26 Morphological Assessments of Traumatic Brain Injury . . . . . . . . . . . . . . . . . . . . . . 257
Daniel van Bregt, Theresa Currier Thomas, Rachel K. Rowe,
and Jonathan Lifshitz