Stimulation and Inhibition of Neurons

333 Pages·2013·7.729 MB·English

by  Stephen Abbott (auth.), Paul M. Pilowsky, Melissa M.J. Farnham, Angelina Y. Fong (eds.)

N
EUROMETHODS
Series Editor
Wolfgang Walz
University of Saskatchewan
Saskatoon, SK, Canada
For further volumes:
http://www.springer.com/series/7657

Stimulation and Inhibition
of Neurons
Edited by
Paul M. Pilowsky, Melissa M.J. Farnham,
and Angelina Y. Fong
Australian School of Advanced Medicine, Macquarie University, North Ryde, NSW, Australia
Editors
Paul M. Pilowsky Melissa M.J. Farnham
Australian School of Advanced Medicine Australian School of Advanced Medicine
Macquarie University Macquarie University
North Ryde, NSW, Australia North Ryde, NSW, Australia
Angelina Y. Fong
Australian School of Advanced Medicine
Macquarie University
North Ryde, NSW, Australia
ISSN 0893-2336 ISSN 1940-6045 (electronic)
ISBN 978-1-62703-232-2 ISBN 978-1-62703-233-9 (eBook)
DOI 10.1007/978-1-62703-233-9
Springer New York Heidelberg Dordrecht London
Library of Congress Control Number: 2012950593
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Series Preface
Under the guidance of its founders Alan Boulton and Glen Baker, the Neuromethods series
by Humana Press has been very successful since the fi rst volume appeared in 1985. In about
17 years, 37 volumes have been published. In 2006, Springer Science + Business Media
made a renewed commitment to this series. The new program will focus on methods either
that are unique to the nervous system and excitable cells or which need special consider-
ation to be applied to the neurosciences. The program will strike a balance between recent
and exciting developments like those concerning new animal models of disease, imaging,
in vivo methods, and more established techniques. These include 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. The
careful application of methods is probably the most important step in the process of scientifi c
inquiry. In the past, new methodologies led the way in developing new disciplines in the
biological and medical sciences. For example, Physiology emerged out of Anatomy in the
nineteenth century by harnessing new methods based on the newly discovered phenome-
non of electricity. Nowadays, the relationships between disciplines and methods are more
complex. Methods are now widely shared between disciplines and research areas. New
developments in electronic publishing also make it possible for scientists to download chap-
ters or protocols selectively within a very short time of encountering them. This new
approach has been taken into account in the design of individual volumes and chapters in
this series.
Wolfgang Walz
v

Preface
Many of our modern investigations into the nervous system are possible because of the
pioneering work from Ludwig’s Institute in the late 1800s ( 1, 2 ) . From this single site came
many of the techniques and discoveries that still provide information on the work being
conducted today. Nevertheless, activation, inhibition or destruction of the nervous system
or its component parts as a vital tool for the investigation of function has undergone remark-
able development in the past 150 years. More recently, new approaches have been devel-
oped that allow for activation, inhibition or destruction of parts of the nervous system as
therapeutic tools. Methods for activating or inhibiting parts of the nervous system have
developed considerably since Borison and Wang’s ( 3 ) initial experiments with Radon lesions
in the medulla oblongata of dogs 60 years ago (Fig. 4 of ( 3 ) ).
The objective of this book is to provide an overview of modern methods for generating
lesions as well as for stimulating and inhibiting neural pathways. Most physiological studies
require activation or inhibition of pathways in order to gain a mechanistic understanding of
sensory inputs, integration and efferent pathways. Currently available approaches provide
sensitivity and specifi city that were unheard of at the time of Borison and Wang ( 3 ) . An
overview of the progress in the use of the neurotransmitter glutamate as a chemical excitant
is provided. This method has its roots in the seminal work of many investigators in the
1970s ( 4 ) . Since that time, investigations into the role of amino acid neurotransmitters
increased dramatically as it became clear that they are responsible for the largest component
of fast neurotransmission in the brain and spinal cord, as well as exerting effects on slowly
acting metabotropic receptors and being required for other cellular activities.
A number of different approaches for activating and inhibiting neurons, nuclei and
nerves are assayed. Many new techniques such as optogenetics and the use of the in situ
perfused preparation are examined. In other sections, the use and validity of more well-
known approaches are reassessed. The use of local anaesthesia to achieve reversible denerva-
tion in physiological studies—an approach that is common in clinical medicine—is examined
in the context of baroreceptor denervation. Less invasive methods of examining barorecep-
tor function using heart rate variability and barorefl ex sensitivity are also considered. At the
intracellular level, cellular and integrative functions resulting from the activation or inhibi-
tion of neuronal G protein systems are assayed. Current approaches to antidromic activa-
tion are then assessed, as is the technique of dye labelling by electroporation of Neurobiotin.
Activation and inhibition of single neurons during intracellular recording as well as using
multibarrel electrodes extracellularly in the vicinity of an intracellular electrode are described.
Analysis of sympathetic nerve activity in the periphery is a topic that has created consider-
able discussion for many years, and this is also examined. Another approach that is com-
monly used in neurophysiology is the compound action potential. Examination of the
compound action potential in relation to vision and estimation of motor units in mice is
vii
vviiiiii Preface
described. Changes in peripheral nerve function following nerve injury and subsequent
neuropathic pain is an important area of study and is discussed in relation to two models.
Finally, a method for improving motor function in humans following stroke is described.
North Ryde, NSW, Australia Paul M. Pilowsky
Melissa M.J. Farnham
Angelina Y. Fong
References
1. F ye WB (1986) Carl Ludwig and the Leipzig Physiological Institute: ‘a factory of new knowledge’.
Circulation 74(5): 920–928
2. L udwig C, Cyon dE (1866) Die Re fl exe eines der sensiblen Nerven des Herzens auf die motorischen
Nerven der Blutgeffässe. Ber der Sächs Ges der Wissenschaften 18: 307–328
3. B orison HL, Wang SC (1951) Quantitative effects of radon implanted in the medulla oblongata: a
technique for producing discrete lesions. J Comp Neurol 94: 35–55
4. W atkins JC, Curtis DR, Felix D (1971) Effect of intracellular injection of pyridine nucleotides on elec-
trical responses of cat spinal motoneurones. Brain Res 35(2): 570–572
Contents
Series Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
PART I NOVEL EXPERIMENTAL TECHNIQUES
1 Optogenetic Stimulation of Neurons in the Brainstem
of Freely Behaving Rats and Mice. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Stephen Abbott
2 Stimulating Peripheral Afferents to Evoke Cardiorespiratory
Reflex Responses in the In Situ Arterially Perfused Preparation. . . . . . . . . . . . . . . . 19
Angelina Y. Fong
3 Excitatory Responses to Microinjection of Glutamate Depend
on Dose Not Volume: A Meta-Analysis of Studies in Rat RVLM. . . . . . . . . . . . . . . 37
Andrea H. Gaede and Paul M. Pilowsky
4 Acute Activation and Inhibition of the Sympathetic
Baroreceptor Reflex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Melissa M.J. Farnham
PART II INTRACELLULAR SIGNALLING
5 Pharmacological Dissection of G Protein-Mediated
Second Messenger Cascades in Neurons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
Mark C. Bellingham
6 In Vivo Manipulation of Intracellular Signalling Pathways. . . . . . . . . . . . . . . . . . . . 107
V.J. Tallapragada
PART III THE SINGLE NEURONS
7 Identification of Spinally Projecting Neurons
in the Rostral Ventrolateral Medulla In Vivo. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
Simon McMullan
8 Microiontophoretic Study of Individual Neurons During
Intracellular Recording. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
Qi-Jian Sun and Paul M. Pilowsky
9 Neurobiotin Electroporation for Combined Structural
and Functional Analysis of Neurons in Developing Mouse Brain Slices . . . . . . . . . . 151
Refik Kanjhan and Mark C. Bellingham
10 Juxtacellular Neuronal Labelling, Physiological Characterization
and Phenotypic Identification of Single Neurons In Vivo . . . . . . . . . . . . . . . . . . . . 167
Anthony J.M. Verberne and Ida J. Llewellyn-Smith
ix