Microchemical Analysis of Nervous Tissue - 1st Edition - ISBN: 9780080181004, 9781483187389

Microchemical Analysis of Nervous Tissue

1st Edition

Methods in Life Sciences

Authors: Neville N. Osborne
eBook ISBN: 9781483187389
Imprint: Pergamon
Published Date: 1st January 1974
Page Count: 240
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Description

Microchemical Analysis of Nervous Tissue focuses on the use of microbiochemical methods in the analysis of nervous tissue, with emphasis on those related to the study of amines, amino acids, phospholipids, and proteins. Special attention is paid to the choice of biological material and the various procedures used for the isolation by dissection of defined components of the nervous system.

Comprised of 10 chapters, this volume begins with an overview of microprocedures used in neurochemistry, followed by a discussion on the importance of choosing the biological material for microanalysis. The isolation of nervous tissue for analysis is then considered, with particular reference to invertebrate neurons; cell components from fresh, "fixed," freeze-dried, and frozen impregnated tissue; and discrete areas of nervous tissue. Subsequent chapters describe some instruments and glassware used in microprocedures, along with the applications of such procedures; general techniques used in microprocedures; microdetermination of phospholipids as well as amines and amino acids as dansyl derivatives; and microelectrophoresis of proteins.

This book will be of interest to molecular biologists, microbiologists, physiologists, and neurochemists.

Table of Contents


Preface

Acknowledgments

1. General Introduction

2. Choice of Biological Material for Microanalysis

3. Isolation of Nervous Tissue for Analysis

Isolation of Characterized Invertebrate Neurons

Isolation of Cell Components from Fresh Tissue

Isolation of Cell Components from 'fixed' Tissue

Isolation of Cell Components from Freeze-Dried Tissue

Isolation of Cell Components from Frozen Impregnated Tissue

Isolation of Discrete Areas of Nervous Tissue

4. Some Instruments and Glassware Used in Microprocedures

Dissecting Tools

Scalpel

Glass Needles

Nylon and Hair Loops

Hair Points

Pipettes

General Considerations

Their Construction

Using the Pipettes

Microtubes

5. Available Microbiochemical Procedures and Their Application

Nucleic Acids

Methods

Studies

Enzyme Activities

Methods

Studies

Ions

Methods

Studies

Transmitter Substances

Screening of Transmitters

Microchromatography of Amines

Fluorometry of Amines

6. General Techniques Used in Microprocedures

Weight Determination

Disruption of Nervous Tissue, e.g. Homogenization

Dialysis

7. Microdetermination of Amines and Amino Acids as Dansyl Derivatives

Introduction

Theoretical and Practical Considerations

The Reaction

Determination of Optimum pH

Incubation Time

Concentration of Dansyl-Chloride

Deductions

Requirements for the Analysis of Tissue Samples

Apparatus and Instruments

Glassware

Chemicals and Solutions

Sundries

General Procedure

Dansyl-Chloride Solution

Extraction of Amines and Amino Acids

Dansylation

Microchromatography

Identification of Amines and Amino Acids on Microchromatograms

Autoradiography and Measurement of Radioactivity

Quantification

Procedure for Analyzing Neurons

Biological Application of Procedure

Amines and Amino Acids in Characterized Neurons

Determination of Contents of Specific Amino Acids and Tryptamine

Detection of Biochemical Change in Nervous Tissue of Minute Dimensions

Other Studies Involving the Use of Microdansyl Procedures

Determination of Amino Acids in Peptides and Proteins

The Possibility of Using Dansyl-Chloride to Detect Noradrenaline, Adrenaline, Octopamine and Related Compounds in the Microscale

8. Microdetermination of Phospholipids

Introduction

Theoretical and Practical Considerations

Advantages of Microchromatography

Preparation of Microchromatography Plates

Chromatography

Identification of Lipids

Quantification

Remarks about Fluorometry

Autoradiography

Microanalysis of Individual Phospholipids in Nervous Tissue

Extraction

Microchromatography

Identification and Elution

Fluorometric Determination

Requirements for the Analysis of Tissue Samples

Apparatus and Instruments

Glassware

Chemicals and Solutions

Sundries

Application of the Procedure

Characterized Cells

Nervous Tissue

9. Microelectrophoresis of Proteins

Introduction

General Principles of Polyacrylamide Electrophoresis

Microelectrophoresis (Partly Homogeneous Polyacrylamide Separating Gel)

General Principle

Requirements

Procedure

Determination of Rf Values of Proteins

Isolation of Individual Proteins

Autoradiography of Microgels

Microelectrophoresis on Gradient Gels

Preparation of Gradient Gels

Fractionation of Proteins

Fractionation of Proteins Containing SDS

Adaptation of Microelectrophoresis

Ribonucleic Acid Species

Analysis of Different Dehydrogenases

Microanalysis of Brain Proteins

Proteins Related to Learning in Nerve Cells

Analysis of Proteins in Invertebrate Neurons

Developmental Changes

Microisoelectric Focusing (LDH)

Principle of the Microisoelectric Focusing Method for Detecting

Reagents

Extracts

Procedure

Demonstration of LDH Patterns

Alternative Buffer System for Microisoelectric Focusing

10. General Comments

Index

Details

No. of pages:
240
Language:
English
Copyright:
© Pergamon 1974
Published:
Imprint:
Pergamon
eBook ISBN:
9781483187389

About the Author

Neville N. Osborne