Neurochemistry: a practical approach
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford [u.a.]
IRL Press
1997
|
| Ausgabe: | 2. ed. |
| Schriftenreihe: | The practical approach series
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XIX, 302 S. Ill., graph. Darst. |
| ISBN: | 0199634408 0199634394 |
Internformat
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Datensatz im Suchindex
| _version_ | 1804144652681478144 |
|---|---|
| adam_text | Neurochemistry
A Practical Approach
Second Edition
Edited by
A J TURNER
Department of Biochemistry and Molecular Biology, University of Leeds,
Leeds LS2 9JT, UK
and
H S BACHELARD
Magnetic Resonance Centre, Department of Physics, University of Nottingham,
University Park, Nottingham NG7 2RD, UK
-at-
OXFORD UNIVERSITY PRESS
Oxford New York Tokyo
Contents
List of contributors xv
Abbreviations xvii
1 Isolation of synaptosomes, growth cones, and
their subcellular components 1
Pauline Phelan and Phillip R Gordon-Weeks
1 Introduction 1
2 Isolation of synaptosomes 3
Introduction 3
Equipment, solutions, and gradients 3
Electron microscopy 9
Assessment of purity 10
3 Isolation of growth cones 11
Introduction 11
Equipment, solutions, gradients, and column chromatography 13
Assessment of purity 17
4 Isolation of the subcellular components of synaptosomes 19
Introduction 19
Isolation of synaptic plasma membranes 19
Isolation of postsynaptic densities 23
Isolation of synaptic vesicles 27
5 Isolation of the subcellular components of growth cones 33
Introduction 33
Equipment, solutions, and gradients 33
Acknowledgements 36
References 36
2 Proteins of the neuronal cytoplasmic and
membrane-associated cytoskeleton 39
Nandini V L Hayes, Fiona E Holmes, Anne Roobol, Martin
J Garden, and Anthony J Baines
1 Introduction 39
2 Microtubules 40
Principles 40
Contents
3 Neurofilaments 49
Principles 49
4 Membrane-associated cytoskeleton: spectrin and synapsin I 55
Principles 55
5 Calmodulin 64
Principles 64
Acknowledgements 66
References 66
3 Immunocytochemical techniques for the
study of the nervous system 71
/ V Priestley
1 Introduction 71
2 Immunocytochemical methods, general background theory
Primary antibodies
Developing antibodies
Purification and storage of immunoreagents
Specificity
Choosing an immunocytochemical method
3 Light microscopic immunocytochemical protocols
General recipes and procedures
Immunofluorescence
Peroxidase-based procedures (PAP and ABC)
Immunogold-silver staining (IGSS)
Double immunocytochemical staining
4 Electron microscopic (EM) immunocytochemistry
General introduction
Pre-embedding staining
Post-embedding staining
5 Combined imm unocytochemistry and in situ hybridization
6 Conclusion
Acknowledgements
References
4 In situ hybridization approaches to the
study of the nervous system 121
K B Seroogy and J P Herman
1 Introduction 121
Contents
2 Procedures 122
Probes 122
Probe labelling 126
Tissue preparation 130
Tissue pre-treatment 134
Hybridization 134
Post-hybridization treatment 135
3 Detection of hybridization signal 136
Film autoradiography 136
Liquid emulsion autoradiography 136
Non-radioactive detection 138
4 Controls 138
5 Double labelling approaches 140
Combined radioactive and non-radioactive in situ hybridization 140
Combined in situ hybridization and immunocytochemistry 141
Combined in situ hybridization and tract-tracing 141
6 Quantitative analysis of in situ hybridization data 142
Quantitation methods 142
Acknowledgements 148
References 148
5 Investigation of mitochondrial defects in
brain and skeletal muscle 151
/ B Clark, T E Bates, P Boakye, A Kuimov, andJ M Land
1 Introduction 151
2 Clinical biochemistry 152
Static tests 152
Dynamic (in vivo function) testing 154
3 Mitochondrial studies 157
Mitochondrial preparation - • 157
Mitochondrial assessment 159
References - 174
6 Measurement of neurotransmitters and
their associated second messengers
in vivo 175
A K Cadogan and C A Marsden
1 Introduction 175
Measurement of neurotransmitters in vivo 175
xi
Contents
Measurement of second messengers in vivo 176
Assays for quantitative analysis of neurotransmitters and second
messengers 176
2 Microdialysis probes 177
Different types of dialysis probe 177
Probe recovery 178
Factors affecting probe recovery 180
3 Perfusion medium and equipment 181
Perfusion fluid 181
Perfusion equipment 181
Perfusate collection 182
Administration of drugs via the probe 183
4 Microdialysis procedures 183
Probe implantation 183
Use of conscious or anaesthetized animals 185
Experiments using the freely-moving animal 188
Effects of microdialysis on the brain microenvironment 190
5 Measurement of neurotransmitters using HPLC-EC 190
Basic principles 190
Amines and their metabolites 191
HPLC-EC equipment 191
Chromatographic conditions for separation of amine
neurotransmitters 192
Acetylcholine 193
Amino acids 194
6 Measurement of second messengers 195
Assays for cAMP and cGMP 195
7 Conclusion 196
Acknowledgements 198
References 198
7 Protein phosphorylation and
dephosphorylation in the nervous system 201
Peter R Dunkley, Paula E Jarvie, and Alistair T R Sim
1 Introduction 201
2 General procedures 202
ATP-labelling of proteins in lysed tissues 204
Prlabelling of proteins in intact tissue 207
Quantitation of protein phosphorylation 212
Safe use of 32P 213
xii
Contents
3 Analysis of substrates
Isolation and identification of phosphoproteins
Characterization of phosphoproteins
Determination of phosphorylation sites
4 Analysis of protein kinases
Content
Activity
5 Analysis of serine/threonine protein phosphatases
Content
Activity
Acknowledgements
References
8 Receptor-linked phosphoinositide
metabolism 229
Ian H Batty, A Nigel Carter, R A John Challiss, and
John N Hawthorne
1 Introduction 229
2 Radiolabelling of phosphoinositides 230
3 Measurement of changes in phosphoinositide metabolism 232
Extraction of phosphoinositides 232
Separation of phospholipids by thin-layer chromatography 235
Detection of phospholipid on thin-layer plates 238
Quantitative analysis of phospholipids on thin-layer plates 239
4 Determination of free and lipid bound inositol 239
5 Measurement of release of inositol phosphates 240
Sample preparation 240
Separation of inositol phosphates by Dowex chromatography 240
Deacylation 241
Analysis by HPLC - 245
6 Mass determination of phosphoinositide cycle
intermediates 252
7 Phosphatidylinositol 3-kinase 255
Assay of PI 3-kinase 256
HPLC separations 260
Measuring PI 3-kinase activation in intact cells 262
Acknowledgements 267
References 267
xiii
Contents
9 Neurotransmitter transporters: purification,
reconstitution, and assay of the brain GABA
transporter 269
Shoshana Keynan
1 Introduction 269
2 The biochemistry and molecular biology of the GABA
transporter 270
3 Purification of neurotransmitter transporters 273
4 Reconstitution of protein into liposomes 275
5 Purification of the GABA transporter 279
6 Conclusions 287
Acknowledgements 288
References 288
Al Suppliers of antibodies 290
A2 Core list of suppliers 294
Index 299
XIV
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| spelling | Neurochemistry a practical approach ed. by A. J. Turner ... 2. ed. Oxford [u.a.] IRL Press 1997 XIX, 302 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series Neurochemistry cabt Neurobiologie (DE-588)4041871-6 gnd rswk-swf Neurochemie (DE-588)4041872-8 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Neurochemie (DE-588)4041872-8 s DE-604 Methode (DE-588)4038971-6 s Neurobiologie (DE-588)4041871-6 s Turner, Anthony J. 1947- Sonstige (DE-588)1043030417 oth HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=021773569&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Neurochemistry a practical approach Neurochemistry cabt Neurobiologie (DE-588)4041871-6 gnd Neurochemie (DE-588)4041872-8 gnd Methode (DE-588)4038971-6 gnd |
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| title | Neurochemistry a practical approach |
| title_auth | Neurochemistry a practical approach |
| title_exact_search | Neurochemistry a practical approach |
| title_full | Neurochemistry a practical approach ed. by A. J. Turner ... |
| title_fullStr | Neurochemistry a practical approach ed. by A. J. Turner ... |
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| title_short | Neurochemistry |
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| title_sub | a practical approach |
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| topic_facet | Neurochemistry Neurobiologie Neurochemie Methode |
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