Table of ContentsLast Updated March, 2002Foreword
by Sir James Black 1 Receptor Binding
1 Receptor Binding1.1 Receptors as Drug Targets1.2 Receptor Theory 1.3 Practical Aspects of Radioligand Binding 1.4 Characterization of Opioid Receptors 1.5 Characterization of Adrenoceptors 1.6 Characterization of Dopamine Receptors 1.7 Characterization of GABA Receptors 1.8 Characterization of Neuronal Nicotinic Acetylcholine Receptors 1.9 Characterization of P1 (Adenosine) Purinoceptors 1.10 Characterization of Angiotensin II Receptors 1.11 Characterization of Neuropeptide Y Receptors 1.12 Characterization of Cholecystokinin Recptors 1.13 Characterization of CRF Receptors 1.14 Characterization of Wild-Type Excitatory Amino Acid Ion Channels Receptors 1.15 Characterization of Tachykinin Receptors 1.16 Characterization of Benzodiazepine Binding to GABAA Receptors 1.17 Characterization of Potassium Channel Binding 1.18 Characterization of the Picrotoxin Site of GABAA Receptors 1.19 Characterization of Histaminergic Receptors 1.20 Radioligand Assay of Drug Activity at the Glycine Site of the NMDA Receptor 1.21 Overview of Receptor Allosterism 1.22 Quantification of Allosteric Interactions at G Protein Coupled Receptors Using Radioligand Binding Assays 1.23 Characterization of 5-HT1A,B and 5-HT2A,C Serotonin Receptor Binding 1.24 Characterization of Chemokine Receptors 1.25 Characterization of Calcium Channel Binding 1.26 Characterization of Cannabinoid Receptors (New, March 2002) 2 Signal Transduction2.1 Overview of Signal Transduction2.2 Adenylate and Guanylate Cyclase Assays 2.3 Overview of Phosphoinositide Hydrolysis 2.4 Nitric Oxide Synthase Assays 2.5 Analysis of Early Gene Responses 2.6 Agonist-Stimulated [35S]GTPgS Binding 2.7 Assay of Receptor-Stimulated Phosphoinositide Turnover 3 Enzyme Assays3.1 Cyclooxygenase Assays3.2 HIV Protease Assays 3.3 Topoisomerase Assays 3.4 Protein Farnesyltransferase Assays 3.5 Protein Tyrosine Kinase Activity Assays 3.6 Monoamine Oxidase Assays 3.7 Characterization of Matrix Metalloproteinase Inhibitors: Enzymatic Assays 3.8 In Vitro Enzymatic Assays of Protein Tyrosine Phosphatase 1B 3.9 Cytochrome P450 Assays (New, December 2001) 3.10 Measurement of Glutamate Carboxypeptidase II (NAALADase) Enzyme Activity by the Hydrolysis of [3H]-N Acetylaspartylglutamate (NAAG) (New, December 2001) 4 Isolated Tissues4.1 Overview of Receptor Interactions of Agonists and Antagonists4.2 Overview of Length-Tension Relationships in Isolated Tissues 4.3 Isolated Cardiac Muscle Assays 4.4 Isolated Blood Vessel Assays 4.5 a- Adrenoceptor Assays 4.6 b- Adrenoceptor Assays 4.7 P1 (Adenosine) Purinoceptor Assays 4.8 Opioid Receptor Assays 4.9 Angiotensin II Receptor Assays 4.10 Tachykinin Receptor Assays 4.11 Neuropeptide Y (NPY) Receptor Assays 4.12 Nicotinic Acetylcholine Receptor Assays 4.13 Cholecystokinin (CKK) Assays 4.14 Dopamine 2 (DA-2) Receptor Assays Using Sympathetic Nerve Terminals 4.15 Muscarinic Receptor Assays 4.16 Cysteinyl Leukotriene Receptor Assays 4.17 Histamine Receptor Assays 4.18 Prostanoid Receptor Assays 5 Animal Models of Disease5.1 Models of Cardiovascular Disease: Measurement of Arrhythmogenic/Antiarrhythmic Activity in the Guinea Pig and Antihypertensive Activity in the Conscious Rat5.2 Models of Pulmonary Disease: Acute and Chronic Allergic Asthma in the Monkey and Acute and Chronic Viral Pulmonitis in the Mouse 5.3 Gastrointestinal Models: Intestinal Transit and Ulcerogenic Activity in the Rat 5.4 Models of Inflammation: Carrageenan-Induced Paw Edema in the Rat 5.5 Models of Inflammation: Adjuvant-Induced Arthritis in the Rat 5.6 Models of Inflammation: Carrageenan Air Pouch in the Rat 5.7 Models of Pain: Hot-Plate and Formalin Tests in Rodents 5.8 Models of Affective Illness: Forced Swimming and Tail Suspension Tests in Rodents 5.9 Affective Illness: Chronic Mild Stress 5.10 Urogenital Dysfunction: Benign Prostatic Hyperplasia 5.11 Models of Urogenital Dysfunction: Urinary Urge Incontinence (UUI) 5.12 Models of Urogenital Dysfunction: Volume-Evoked Micturition Reflex (VEMR) in the Conscious Rat 5.13 Models of Dimentia: Delayed Alternation in Aged Rats 5.14 Spatial Discrimination Water Maze Test in Septal-Lesioned Rats 5.15 Models of Schizophrenia: PPI of Startle in Rats 5.16 Models of Anxiety: Stress-Induced Hyperthermia in Group-Housed Mice 5.17 Models for Assessing Antipsychotics: Amphetamine-Induced Hyperactivity and Stereotypies in Mice 5.18 Models of Anxiety: Ultrasonic Vocalizations of Isolated Rat Pups 5.19 Models for Environmentally Induced Eating Disorders: Dietary Hyperphagia and Anorexia Nervosa 5.20 Overview of the Use of Transgenic Animals in CNS Drug Discovery 5.21 Models of Renal Insufficiency: The Anti-Thy-1.1 Model of Acute Proliferative Glomerulonephritis 5.22 Models of Epilepsy: Electroshock and Chemical Induced Convulsions in the Mouse 5.23 Characterization of Matrix Metalloproteinase Inhibitors: Angiogenesis and Tumor Models 5.24 Classic In Vivo Cancer Models: Three Examples of Mouse Models Used in Experimental Therapeutics (New, December 2001) 5.25 Overview of Animal Models of Asthma (New, March 2002) 5.26 Guinea Pig Models of Asthma (New, March 2002) 5.27 Models of Cardiac Ischemia-Reperfusion Injury in Dogs and Rats (New, March 2002) 6 Molecular Biology6.1 Overview of Receptor Cloning6.2 Receptor Reporter Systems 6.3 Expression of Cloned Receptors in Mammalian Cell Lines 6.4 Isotopic Assays for Reporter Gene Activity 6.5 Nonisotopic Assays for Reporter Gene Activity 6.6 Use of the A. Victoria Green Fluorescent Protein to Study Protein Dynamics In Vivo 6.7 Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization 6.8 Mobility Shift DNA-Binding Assay Using Gel Electrophoresis 7 Pharmacokinetics7.1 Overview of Pharmacokinetics7.2 Estimating Intestinal Mucosal Permeation of Compounds Using Caco-2 Cell Monolayers 7.3 Overview of Drug Product Development (New, December 2001) 8 Receptor/Enzyme Localization8.1 In Vitro Autoradiography8.2 Autoradiographic Localization of Growth Factor Receptors in Neuronal Tissues 9 Drug Discovery Technologies9.1 Overview of Chemical Diversity9.2 Cell-Based Assays Using the Fluorometric Imaging Plate Reader (FLIPR) 9.3 Overview of Combinatorial Chemistry 9.4 Overview of High-Throughput Screening 9.5 Screening in Constitutively Active Receptor Systems 10 Safety Pharmacology/Toxicology10.1 Overview of Safety Pharmacology10.2 Models of Inflammation: Gastrointestinal Ulceration in the Rat 10.3 Toxicology in Drug Discovery and Development 10.4 Place Preference Test in Rats 10.5 Substance Abuse: Self-Administration in Monkeys 10.6 Vigilance-Controlled Quantified EEG in Safety Pharmacology 10.7 An Overview of QT Interval Assessment in Safety Pharmacology 11 Electrophysiological Techniques11.1 Overview of Electrophysiology11.2 Electrophysiological Analysis of G Protein-Coupled Receptors in Mammalian Neurons 11.3 The Action Potential of the Purkinje Fiber: An In Vitro Model for Evaluation of the Proarrhythmic Potential of Cardiac and Noncardiac Drugs (New, December 2001) 12 In Vitro Cellular Assays12.1 Overview of Cell and Tissue Culture Techniques12.2 Chondrocyte Culture and Assay 12.3 Preparation of Nuclear and Cytoplasmic Extracts from Mammalian Cell 12.4 Cellular Assays of Chemokine Receptor Activation AppendicesA.1 NOMENCLATURE AND USEFUL DATAA.2 STOCK SOLUTIONS, EQUIPMENT, AND LABORATORY
GUIDELINES A.3 STANDARD TECHNIQUES |