Muscarinic Receptor: From Structure to Animal Models

294 Pages·2016·10.274 MB·English

by  Jaromir Myslivecek, Jan Jakubik (eds.)

Neuromethods 107
Jaromir Myslivecek
Jan Jakubik Editors
Muscarinic Receptor:
From Structure
to Animal Models
N
EUROMETHODS
Series Editor
Wolfgang Walz
University of Saskatchewan
Saskatoon , Canada
For further volumes:
http://www.springer.com/series/7657

Muscarinic Receptor:
From Structure to Animal Models
Edited by
Jaromir Myslivecek
Institute of Physiology, Charles University, Prague, Czech Republic
Jan Jakubik
Department of Neurochemistry, Acedemy of Sciences of the Czech Republic, Institute of Physiology,
Prague, Czech Republic
Editors
Jaromir Myslivecek Jan J akubik
Institute of Physiology Department of Neurochemistry
Charles University Academy of Sciences of the Czech Republic
Prague, Czech Republic Institute of Physiology
Prague, Czech Republic
ISSN 0893-2336 ISSN 1940-6045 (electronic)
Neuromethods
ISBN 978-1-4939-2857-6 ISBN 978-1-4939-2858-3 (eBook)
DOI 10.1007/978-1-4939-2858-3
Library of Congress Control Number: 2015945343
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Prefa ce
Muscarinic receptors are involved in a number of physiological events like cognitive
processes, motor coordination, attention, circadian rhythms, food reinforcement, drug
addiction, and synaptic plasticity. This book provides methodology for the study of musca-
rinic receptors at the structural to systemic level. The chapters are primarily intended as a
resource for scientists who want to newly establish protocols to study muscarinic receptors
without hitches and falling to potential pitfalls. One of such pitfalls is the lack of subtype-
selective ligands that makes studies targeted to specifi c subtype problematic. One of the
methodological approaches for subtype identifi cation in tissues and organs is immunohis-
tochemical or Western blot analysis of muscarinic receptors. However, these methods have
strong limitations as the selectivity of antibodies is usually poor and antibodies also target
nonfunctional and degraded receptors. Thus these methods do not provide assessment of
real number of receptors. Moreover, some artifacts can originate from tissue preparation.
These can be avoided by studying receptors in their natural environment.
We start our book with methods for characterization of muscarinic receptor in crystal-
lography studies that advanced our understanding of structural properties and activation
mechanism of muscarinic receptors and are cornerstone in molecular modeling and
computer- based approaches to study muscarinic receptors. Then we move to binding tech-
niques that thanks to heterologous expression systems allow us to perform binding studies
very accurately and easily and discuss overcoming diffi culties arising from the lack of selec-
tive ligands. Next we provide protocols to investigate molecular properties of muscarinic
receptors. Then we provide protocols to study muscarinic receptors in the central nervous
system using autoradiography and PET studies. We end with protocols on animals with
knock-out and knock-in muscarinic genes to study the role of muscarinic receptors in physi-
ology and behavior.
Prague, Czech Republic Jaromir Myslivecek
Jan J akubik
v

Series Preface
Experimental life sciences have two basic foundations: concepts and tools. The N euromethods
series focuses on the tools and techniques unique to the investigation of the nervous system
and excitable cells. It will not, however, shortchange the concept side of things as care has
been taken to integrate these tools within the context of the concepts and questions under
investigation. In this way, the series is unique in that it not only collects protocols but also
includes theoretical background information and critiques which led to the methods and
their development. Thus it gives the reader a better understanding of the origin of the
techniques and their potential future development. The N euromethods publishing program
strikes a balance between recent and exciting developments like those concerning new ani-
mal models of disease, imaging, in vivo methods, and more established techniques, includ-
ing, for example, immunocytochemistry and electrophysiological technologies. New
trainees in neurosciences still need a sound footing in these older methods in order to apply
a critical approach to their results.
Under the guidance of its founders, Alan Boulton and Glen Baker, the N euromethods
series has been a success since its fi rst volume published through Humana Press in 1985.
The series continues to fl ourish through many changes over the years. It is now published
under the umbrella of Springer Protocols. While methods involving brain research have
changed a lot since the series started, the publishing environment and technology have
changed even more radically. Neuromethods has the distinct layout and style of the Springer
Protocols program, designed specifi cally for readability and ease of reference in a laboratory
setting.
The careful application of methods is potentially the most important step in the process
of scientifi c inquiry. In the past, new methodologies led the way in developing new disci-
plines in the biological and medical sciences. For example, Physiology emerged out of
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ered phenomenon of electricity. Nowadays, the relationships between disciplines and meth-
ods are more complex. Methods are now widely shared between disciplines and research
areas. New developments in electronic publishing make it possible for scientists that
encounter new methods to quickly fi nd sources of information electronically. The design of
individual volumes and chapters in this series takes this new access technology into account.
Springer Protocols makes it possible to download single protocols separately. In addition,
Springer makes its print-on-demand technology available globally. A print copy can there-
fore be acquired quickly and for a competitive price anywhere in the world.
Wolfgang W alz
vii

Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Series Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v ii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x i
1 Towards the Crystal Structure Determination of Muscarinic
Acetylcholine Receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Ryoji S uno , Hidetsugu A sada , and Takuya K obayashi
2 H omology Modeling and Docking Evaluation of Human Muscarinic
Acetylcholine Receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 5
Trayder Thomas , D avid K . C halmers , and E lizabeth Yuriev
3 Radioligand Binding at Muscarinic Receptors . . . . . . . . . . . . . . . . . . . . . . . . . 3 7
Esam E . El-Fakahany and Jan J akubik
4 B inding Method for Detection of Muscarinic Acetylcholine Receptors
in Receptor’s Natural Environment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Ikunobu Muramatsu , Hatsumi Y oshiki , Kiyonao S ada ,
Junsuke U wada , Takanobu T aniguchi , Takayoshi Masuoka ,
and Matomo N ishio
5 Use of Antibodies in the Research on Muscarinic Receptor Subtypes. . . . . . . . 8 3
Wisuit P radidarcheep and M artin C. M ichel
6 A llosteric Modulation of Muscarinic Receptors . . . . . . . . . . . . . . . . . . . . . . . . 9 5
Jan Jakubik and Esam E . E l-Fakahany
7 Subcellular and Synaptic Localization of Muscarinic Receptors
in Neurons Using High-Resolution Electron Microscopic
Preembedding Immunogold Technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 31
Véronique B ernard
8 Investigation of Muscarinic Receptors by Fluorescent Techniques . . . . . . . . . . 147
Cornelius K rasel , Andreas R inne , and Moritz Bünemann
9 A utoradiography Assessment of Muscarinic Receptors
in the Central Nervous System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 59
Vladimir F arar and J aromir M yslivecek
10 Imaging of Muscarinic Receptors in the Central Nervous System. . . . . . . . . . . 181
Hideo T sukada , S hingo Nishiyama , and Kazuhiro T akahashi
11 D etection of Non-neuronal Acetylcholine. . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
Ignaz Karl Wessler and Charles James Kirkpatrick
12 U tilization of Superfused Cerebral Slices in Probing Muscarinic
Receptor Autoregulation of Acetylcholine Release. . . . . . . . . . . . . . . . . . . . . . 221
Glenda Alquicer, Vladimír D oležal and Esam E. E l-Fakahany
ix