Practical Pharmacology for the Pharmaceutical Sciences

Practical Pharmacology for the Pharmaceutical Sciences is a lab survival guide for those studying Pharmacology, providing hands-on advice on developing pharmacology laboratory and data handling skills. Suitable for both undergraduates and postgraduates, it focuses on laboratory techniques rather than computer-simulated data. It also guides the reader through the process of communicating experimental results in a variety of formats, including posters, oral presentations and project reports.

Split into three main areas, the following topics are covered in detail: 

Preparation for Experimental Pharmacology

• Legal aspects
• Fundamentals of Pharmacology
• Definitions, calculations and statistics

Experiments in Pharmacology

• Microtitre-based techniques using isolated cells
• In vitro techniques using isolated tissues and organs
• Biochemical techniques using cell-free systems

Communicating experimental results

• Data presentation
• How to write scientific reports
• Pharmacological literature

Supported with numerous questions throughout the text, as well as step by step instructions for practical experiments, this book presents an approach to learning pharmacology through an appreciation of authentic experimental data.

Dr D. Michael Salmon, School of Health and Biosciences, University of East London
Dr Salmon studied for a Ph.D. in Biochemistry at Imperial College of Science and Technology, London. He has carried out research on signal transduction at U. Mass. Medical School, USA, National Institute for Biological Standards and Control, London and St. George's Hospital Medical School, London. Since 1989, he has taught and researched in pharmacology at the University of East London. He has been an author on 24 scientific papers in peer-reviewed journals, presented 31 communications to Learned Societies and contributed to 8 books and scientific monographs. He has also contributed to The British Pharmacological Society sponsored computer learning program for pharmacology, pharma-CAL-ogy.

Preface ix

Acknowledgements xi

1 Before Entering the Pharmacology Laboratory 1

1.1 Safety and Risk Assessment 1

1.2 The Laboratory Record Book 3

1.3 Use of Animals in Practical Pharmacology 4

1.4 Experimental Design 5

1.5 Units, Dilutions and Logarithms 7

1.5.1 Units of Mass 8

1.5.2 Units, Concentrations and Logarithms 8

1.5.3 Dilutions 9

1.5.4 Logarithms 10

1.6 Essential Statistics 12

1.6.1 Continuous Data – t-test, ANOVA,

Non-parametric Tests and Regression 12

1.6.2 Discontinuous Data – 2 and Fisher’s Exact Test 21

References 25

2 Basic Pharmacological Principles 27

2.1 Drug–receptor Interaction 27

2.1.1 Agonists 27

2.1.2 Antagonists 30

2.1.3 Receptor Classification 36

2.2 Bioassays 37

2.2.1 Single-point Assays 38

2.2.2 Bracketing Assays, Three-point or 2×1 Assays 38

2.2.3 Multi-point Assays, Such As Four-point or

2×2 Assays 39

References 40

3 Isolated Tissues and Organs 43

3.1 Equipment for In Vitro Experiments 44

3.2 Organ Baths 45

3.3 Physiological Salt Solutions 46

3.4 Transducers 47

3.5 Recording Equipment and Software 49

3.6 Dosing 50

3.7 Electrically Stimulated Preparations 52

3.8 Fault-Finding of In Vitro Isolated Tissue Preparations 53

References 54

4 Smooth Muscle Preparations 55

4.1 Gastrointestinal Smooth Muscle Preparations 55

4.2 Guinea Pig Isolated Ileum 56

4.2.1 Concentration–Response Curves

for Cholinesters 57

4.2.2 Selective Antagonism 59

4.2.3 Specificity of Blood Cholinesterases 62

4.2.4 Quantification of the Potency of an Antagonist 64

4.2.5 Bioassays 67

4.2.6 Calcium Channel Blockers 73

4.2.7 Field-stimulated Guinea Pig Isolated Ileum 76

4.3 Rabbit Isolated Jejunum and the Finkleman Preparation 78

4.3.1 Adrenoceptor Sub-types 79

4.4 Isolated Tracheal Rings 80

4.5 Isolated Vas Deferens 82

Questions on Isolated Tissue Preparations 83

Answers to Problems 87

References 92

5 Cardiovascular Preparations 93

5.1 Isolated Perfused Heart Preparations 94

5.1.1 The Langendorff Preparation 95

5.1.2 Cardiac Interactions of Anti-asthma Drugs 98

5.1.3 The Rat Isolated Auricle Preparation 99

5.2 Thoracic Aorta Preparation 102

5.2.1 Drugs Regulating Nitric Oxide-mediated

Relaxation 104

References 105

6 Skeletal Muscle 107

6.1 Types of Skeletal Muscle 107

6.2 Multiply-Innervated Skeletal Muscle Preparations 108

6.2.1 Agonists and Antagonists Acting on the Frog

Rectus Abdominis 109

6.2.2 Action of Anticholinesterases on the Dorsal

Muscle of the Leech 111

6.3 Focally Innervated Skeletal Muscle Preparations 116

6.3.1 The Frog Gastrocnemius Muscle–Sciatic

Nerve Preparation 119

References 120

7 Isolated Cells 121

7.1 Freshly Isolated and Cultured Cells 121

7.1.1 Advantages of Isolated Cells 121

7.1.2 Cultured Cells 122

7.1.3 Cell Counting 122

7.2 Platelets 125

7.2.1 Inhibition of Aggregation by Nitric Oxide

Donors 127

7.3 Neutrophils 131

7.3.1 Measurement of NADPH Cytochrome c

Reductase 132

7.3.2 Measurement of Intracellular [Ca2+] 134

References 139

8 Biochemical Pharmacology 141

8.1 Pharmacological Applications of Common Biochemical

Techniques 141

8.2 Enzyme Inhibitors 142

8.3 Acetylcholinesterase Inhibitors 143

8.4 Monoamine Oxidase Inhibitors 145

8.4.1 Sub-cellular Distribution of MAO Activity 146

8.4.2 Specificity of MAO Inhibitors for Isoenzymes 149

8.5 Thrombin Inhibitors 151

8.6 ATPase Inhibitors 155

References 158

9 Complementary Methods for Teaching Practical

Pharmacology 161

9.1 The Comparative Merits of Available Methods 161

9.2 Interpretation of Experimental Data 162

9.2.1 Behavioural Experiments 162

9.2.2 Analysis of Metabolites of

5-hydroxytryptamine 166

9.2.3 Radioligand Binding 167

Answers to Questions 171

References 176

10 Communicating Results 177

10.1 Preliminary Reports 177

10.1.1 Tables 178

10.1.2 Graphs 178

10.1.3 Bar Graphs 179

10.1.4 Preliminary Conclusions 179

10.2 Poster Presentations 180

10.3 Oral Presentations 181

10.4 Project Reports 183

10.5 Pharmacological Literature 184

10.6 How to Cite Scientific Information Sources 187

10.7 Plagiarism 188

References 188

Appendix 189

Index

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