Methods in pain research lawrence kruger, CRC press, 2001 scan

METHODS IN PAIN RESEARCH 0035-FM-frame Page Tuesday, May 8, 2001 10:16 AM METHODS & NEW FRONTIERS IN NEUROSCIENCE Series Editors Sidney A Simon, Ph.D Miguel A.L Nicolelis, M.D., Ph.D Published Titles Apoptosis in Neurobiology Yusuf A Hannun, M.D., Professor/Biomedical Research and Department Chairman/ Biochemistry and Molecular Biology, Medical University of South Carolina Rose-Mary Boustany, M.D., tenured Associate Professor/Pediatrics and Neurobiology, Duke University Medical Center Methods for Neural Ensemble Recordings Miguel A.L Nicolelis, M.D., Ph.D., Associate Professor/Department of Neurobiology, Duke University Medical Center Methods of Behavioral Analysis in Neuroscience Jerry J Buccafusco, Ph.D., Professor/Pharmacology and Toxicology, Professor/Psychiatry and Health Behavior, Medical College of Georgia Neural Prostheses for Restoration of Sensory and Motor Function John K Chapin, Ph.D., MCP and Hahnemann School of Medicine Karen A Moxon, Ph.D., Department of Electrical and Computer Engineering, Drexel University Computational Neuroscience: Realistic Modeling for Experimentalists Eric DeSchutter, M.D., Ph.D., Department of Medicine, University of Antwerp 0035-FM-frame Page Tuesday, May 8, 2001 10:16 AM METHODS IN PAIN RESEARCH Edited by Lawrence Kruger Professor of Neurobiology (Emeritus) University of California, Los Angeles (UCLA) School of Medicine Los Angeles, CA CRC Press Boca Raton London New York Washington, D.C CRC Press Taylor & Francis Group 6000 Broken Sound Parkway NW, Suite 300 Boca Raton, FL 33487-2742 © 2001 by Taylor & Francis Group, LLC CRC Press is an imprint of Taylor & Francis Group, an Informa business No claim to original U.S Government works Version Date: 20140124 International Standard Book Number-13: 978-1-4200-4256-6 (eBook - PDF) This book contains information obtained from authentic and highly regarded sources While all reasonable efforts have been made to publish reliable data and information, neither the author[s] nor the publisher can accept any legal responsibility or liability for any errors or omissions that may be made The publishers wish to make clear that any views or opinions expressed in this book by individual editors, authors or contributors are personal to them and not necessarily reflect the views/opinions of the publishers The information or guidance contained in this book is intended for use by medical, scientific or health-care professionals and is provided strictly as a supplement to the medical or other professional’s own judgement, their knowledge of the patient’s medical history, relevant manufacturer’s instructions and the appropriate best practice guidelines Because of the rapid advances in medical science, any information or advice on dosages, procedures or diagnoses should be independently verified The reader is strongly urged to consult the drug companies’ printed instructions, and their websites, before administering any of the drugs recommended in this book This book does not indicate whether a particular treatment is appropriate or suitable for a particular individual Ultimately it is the sole responsibility of the medical professional to make his or her own professional judgements, so as to advise and treat patients appropriately The authors and publishers have also attempted to trace the copyright holders of all material reproduced in this publication and apologize to copyright holders if permission to publish in this form has not been obtained If any copyright material has not been acknowledged please write and let us know so we may rectify in any future reprint Except as permitted under U.S Copyright Law, no part of this book may be reprinted, reproduced, transmitted, or utilized in any form by any electronic, mechanical, or other means, now known or hereafter invented, including photocopying, microfilming, and recording, or in any information storage or retrieval system, without written permission from the publishers For permission to photocopy or use material electronically from this work, please access www.copyright com (http://www.copyright.com/) or contact the Copyright Clearance Center, Inc (CCC), 222 Rosewood Drive, Danvers, MA 01923, 978-750-8400 CCC is a not-for-profit organization that provides licenses and registration for a variety of users For organizations that have been granted a photocopy license by the CCC, a separate system of payment has been arranged Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only for identification and explanation without intent to infringe Visit the Taylor & Francis Web site at http://www.taylorandfrancis.com and the CRC Press Web site at http://www.crcpress.com 0035-FM-frame Page Tuesday, May 8, 2001 10:16 AM Methods & New Frontiers in Neuroscience Series Editors Sidney A Simon, Ph.D Miguel A.L Nicolelis, M.D., Ph.D Our goal in creating the Methods & New Frontiers in Neuroscience Series is to present the insights of experts on emerging experimental techniques and theoretical concepts that are, or will be, at the vanguard of neuroscience Books in the series cover topics ranging from methods to investigate apoptosis to modern techniques for neural ensemble recordings in behaving animals The series also covers new and exciting multidisciplinary areas of brain research, such as computational neuroscience and neuroengineering, and describes breakthroughs in classical fields such as behavioral neuroscience We want these to be the books every neuroscientist will use in order to get acquainted with new methodologies in brain research These books can be given to graduate students and postdoctoral fellows when they are looking for guidance to start a new line of research Each book is edited by an expert and consists of chapters written by the leaders in a particular field Books are richly illustrated and contain comprehensive bibliographies Chapters provide substantial background material relevant to the particular subject Hence, they are not just “methods books.” They contain detailed “tricks of the trade” and information as to where these methods can be safely applied In addition, they include information about where to buy equipment and websites that are helpful in solving both practical and theoretical problems We hope that as the volumes become available the effort put in by us, the publisher, the book editors, and individual authors will contribute to the further development of brain research The extent that we achieve this goal will be determined by the utility of these books 0035-FM-frame Page Tuesday, May 8, 2001 10:16 AM 0035-FM-frame Page Thursday, November 15, 2001 8:33 AM Preface The invitation to assemble a book on methodology in pain research proffered by my longtime colleagues, Sid Simon and Miguel Nicolelis, was initially received with considerable uncertainty and hesitation “Pain” encompasses a rather vast field of research and is hardly a subject that could be reduced to a few formulaic chapters describing laboratory protocols ranging from surgical techniques to automated molecular biology, each of which could encompass an entire volume and still prove inadequate for the needs of investigators seeking methodological guidance in this field of endeavor Discussions with several friends whose research interests are deeply committed to studying pain persuaded me to recognize that this is no longer an era of publishing lab cookbooks that would be largely out of date by the time they emerge from the printing press Besides, we are now in an epoch in which new technologies are being developed at a terrifying pace in such areas as high throughput methods, chemical genomics, microarray applications, genotyping, and proteomics The modern investigator keeps current on such rapidly advancing subjects via the World Wide Web, and protocols generally are supplied by vendors of apparatus and reagents But pain research, despite its vastness, is something quite different and special; in part because of its breadth, but largely because it touches upon a subjective report in humans that is poorly understood and yet of fundamental importance and interest to everyone Pain remains the singular most common complaint of patients worldwide, and this is followed by the next most prevalent affliction — the related phenomenon of itch Yet pain has not reached the status of a targeted “disease;” it is clearly part of ordinary human experience It stands apart from other sensations by often being dissociated from a distinctive stimulus condition; i.e., pain can be elicited by apparently innocuous stimuli in pathology or when applied to previously injured tissues Thus, pain can be described pragmatically as a response to a variety of stimuli and contextual circumstances Unlike other sensations, it can be altered significantly by widely known drugs, many of which are “controlled substances,” imposing serious societal problems and consequent encumbrances for laboratory study Finally, it should be emphasized that specific nerve fibers, now called “nociceptors,” were disavowed explicitly over much of the last few decades Now that laboratory experiments can reveal and enable investigators to parse stimulus and response variables, the field has attracted profoundly expanded interest Many of the relevant methods are employed in other areas of neuroscience research and some of the specific techniques are covered in several excellent monographs in the CRC Press series joined by this volume We have attempted here to select topics and authors to meet the needs of investigators seeking guidance on the “how” and the “means” for studying the idiosyncratic features of pain research Major areas of contemporary efforts are outlined and explained, and sources are 0035_frame_IDX Page 305 Wednesday, May 2, 2001 7:00 AM Index 129 mice, 47, 58, 59 A Abdominal constriction (writhing) test, 26–27, 76 Aβ nociceptors, 268, 269, 286 in CCI testing, 81 ACC (Anterior cingulate cortex), 227–228, 232 activation and unpleasantness, 228 MEG studies, 234 pain related activations, 235 Acetaminophen, 288 Acetic acid, 96, 99 Acetone, 99 Acetylcholine measurement by detector patch method, 139 by microdialysis, 136 6-Acetylmorphine, 48 Acid pain stimulation in human subjects, 268 Acrolein, 102 Acrolein fixative, 198 Acute pain definition, 71, 242 fMRI studies, 247 model comparison, 82 models, 71–76; see also specific tests models dissociated from tonic pain models, 15 Aδ-fiber mediated first pain, 268 Aδ-mechano-heat skin nociceptors, 267 Aδ nociceptors, 268 affected by formalin, 27–28 in CCI testing, 81 electrical stimulation mediation, 288 inhibition by opioid agonists, 16 noxious heat stimuli threshold, 18 reflex response, 278 sensations, 269 threshold in mice, 23 Adenosine, 287, 288 effects on heat pain threshold, 286 measurement, 136 AFM (Atomic force microscopy), 140 Alfentanil, 287 Algogens, inflammatory, 16, 272–273 Allodynia defined, 76 Alzheimer's disease, 110, 227, 252 AMCA staining, 192 American Pain Society (APS), Amines, biogenic, 136–137 γ-Aminobutyric acid (GABA) detection by immunocytochemistry, 188; see also GABA AMPA detection by immunocytochemistry, 188 Analgesia definition, experimental measurement factors, 15 Analgesic profiles, 285 Anatomical methods in pain research, 187–207 Anesthesia dolorosa, 79–81 definition, 79–80 Animal research guidelines, 3, 71 Animals; see also Mice; Rats cats, 99 dietary effects on responses, 82–83 dogs, 96 habituation, 14 handling and stress, 13–14 horses, 96 knockout studies restricted to mice, 12 methods for measuring pain in, 71 mice used for genetic studies, mice vs rats as experimental subjects, 13–14 neophobia, 14 nociception tests, see specific tests pain models, 67–87; see also specific models pain response similarity to humans, 68–71 primates, 99 rabbits, 96 rats as principal laboratory subject, rats vs mice as experimental subjects, 13–14 regulation of use in research, species used in pain research, 5, 12t as test subjects, Animal studies pain inferred from behaviors, 11–12 postherpetic neuralgia, 84–85 Anterior cingulate cortex (ACC), see ACC Anterior insula, 227–228, 232 Antibodies commercially available, 119, 120t detection by immunocytochemistry, 194 specificity of commercially available, 119 Antibody microprobes, 137, 139 Apamin resistant current, 180 Aplysia neurons in neuropathic pain research, APS (American Pain Society), 305 0035_frame_IDX Page 306 Wednesday, May 2, 2001 7:00 AM 306 Araldite embedding medium preparation, 207 Astrocytes, expression of MHC II, 124 Atomic force microscopy (AFM), 140 ATP measurement by AFM, 140 by detector patch method, 139 luciferin/luciferase method, 140 Autotomy, 80–81 Avidin conjugates, 192, 195, 197, 200, 201 Axon fiber strand measurements, 151 B BAC (bacteria artificial chromosomes), 57 Backcrossing techniques, 58–59 Back pain, 242, 249–251 brain regional biochemistry, 254–255 chronic alters brain chemistry, 256 fMRI studies, 246 lumbar radiculopathy, 113 with radicular involvement, 250 time variation by fMRI, 251 Baclofen, 101 Bacteria artificial chromosomes (BAC), 57 BALB mice, 47, 59 Bandeiraea simplicifolia isolectin B4 (BSIB4), 197 Basal ganglia, 228, 232 BCA (bicinchoninic acid) assay, 121 BCIP/NBT detection, 115 Benzodizapine receptor binding studied by PET imaging, 227 Bicinchoninic acid (BCA) assay, 121 Biocytin tracer, 201 Bioinformatics, 8–9 BioMed Central, Biowire, Bladder irritation models, 94 Blister suction sampling, 134 Bloc staining, 199 BOLD (Blood oxygenation level dependent) effect, 230, 244 Bradykinin, 272, 273 evoked responses, 174 as inflammatory mediator, 172 receptor antagonists, 101 Brain biochemistry measurement by MRS, 251–258; see also specific species subjective pain reflected in, 256–257 Brain hemodynamics, 246 Brain imaging in human subjects, 225–235 choice of methods, 234 chronic pain pathophysiology, 241–259 Brain tumors, 252 Methods in Pain Research Brainstem sampling by push-pull cannula, 135 (BSIB4) Bandeiraea simplicifolia isolectin B4, 197 BuBu agonist, 51 C C3H/He mice, 21 C57BL/6 mice, 21, 30, 47, 58, 59 avoidance learning, 59 Ca+2 concentration determination at membranes, ion channels, 171 measurement by ISM, 138 Ca+2 currents, 176, 178–179 Cl- current dependence, 181–182 K+ current dependence, 180–181 Cadherins, 125 Caerulein, 98 Caffeine, 288 Calcitonin gene-related peptide, see CGRP CAM (Cellular adhesion molecules), 125–126 Cantharidin-induced blisters, 134 CAP (Compound action potential recording), 149–150 Capsaicin, 99, 101, 272, 287 Capsaicin pain stimulation in human subjects, 268 Capsaicin receptor cloning, Capsaicin response C-fiber-mediated, 16 fMRI studies, 233 Capsaicin sensitivity, 126 Capsaicin stimulation in human subjects, 269–270 Carbamazepine, 83 Carbocyanine dyes, 202–203 Carbon fiber electrode method, 136–137 Carrageenan, 16, 112 CAT (Chloramphenicol acetyltransferase) reporter, 45–46 Cats, 99 11C by PET imaging, 227 CCI (Chronic constriction injury), 83, 111, 112 methods, 81 testing, 74–75 CD8 expression, 124 13C determination in vivo in the brain by MRS, 252 Cecal distension, 96 Cell body as model for afferent terminal, 170–172 Cell-attached patch, see Patch-clamp recording Cellular adhesion molecules (CAM), 125–126 intracellular (ICAM), 125, 126 platelet (PECAM), 125, 126 0035_frame_IDX Page 307 Wednesday, May 2, 2001 7:00 AM Index Central pain, 79 Central pain syndromes, 282–283 Central sensitization, 112, 269, 273, 283 analgesic drugs for cutaneous pain, 287 by capsaicin, 270 in chemical tests, 28 glial cells, 123–124 immunocompetent glia, 123–124 Central summation of nociceptive activity in human subjects, 268 Cerebellum, 228, 232 Cerebral blood flow (CBF) mapping, 229 studies by PET imaging, 227 C-fiber mediated second pain, 268 C-fiber nociceptors, 149, 286 C-fibers, 81, 84, 268 affected by formalin, 27–28 capsaicin responses, 16 function, 287 ion channel activity recording, 156–157 noxious heat stimuli threshold, 18 in pain sensation, 150–151 saline-induced muscle pain mediation, 288 sensations, 269 sensitization, 111 threshold in mice, 23 C-fos proteins, 102 CGRP (Calcitonin gene-related peptide), 272 immunocytochemistry, 194 measurement in vivo, 137 Charybdotoxin-sensitive current, 180 Chemical assays, 25–29 Chemical nociceptive assays, see specific tests suprathreshold, 16 Chemical pain stimulation in human subjects, 268, 272–273 Chemosensitivity of neurons, 172 Chemotherapy-evoked peripheral neuropathy, 85–87, 102 Chicken pox, 84–85 CHIP (cDNA homologous insertion protocol), 56 Chloramphenicol acetyltransferase (CAT) reporter, 45–46 Cholecystokinin in brain and spinal cord, 136 Cholera toxin B subunit tracer, 196, 201 Choline acetyltransferase detection, 254 by immunocytochemistry, 188 Choline determination in vivo in the brain by MRS, 252 Chronic constriction injury (CCI), see CCI Chronic pain, 2, 71, 243 arthritis models, 77–78 definition, 71, 242 fMRI studies, 233, 247 307 freeze lesions, 113 genetic role, 124–125 important brain structures, 258 measurement difficulties, 69 models, 77–79 neural degeneration in, 258 PET studies, 228–229 SPECT imaging, 229–230 study of mRNA nodulation, 116–117 Cingulate, 246, 256, 257 in back pain, 254 pain related activations, 235 Cisplatin, 85 CiteTrack, Clinical pain, see Chronic pain Cl- measurement by ISM, 138 Cluster headache SPECT imaging, 229 C-mechano-heat skin nociceptors, 267 (CMV) Human cytomegalovirus for transgenesis, 44 CNS (Central nervous system), immune system functioning in, 123–126 Cocaine reward blocking, 53 Cold-allodynia tests, 75 Cold pain stimulation in human subjects, 267 Cold-plate test, 22 Colorectal distension (CRD) methods, see CRD Colorectal distension in the rat, see CRD Complete Freund's adjuvant, see Freund's adjuvant Complex Regional Pain Syndrome I, 125 Compound action potential (CAP) recording, 149–150 Conconavalin-A in plating substrate, 159 Conduction velocity, 149 Confocal laser scanning microscopy, 192, 197, 201 in immunocytochemistry, 194 Confounds increase with increased test sensitivity, 17 Contracture-induced pain, 280 Controlled substance legal issues, Coronary artery occlusion, 103 Coronary artery occlusion model, 94 Cortex activations by MEG studies, 233 Cortical activations in pain response, 228, 247–248 Cortical cup method, 134 Corticosteroids, response in bowel distension, 97 Cosmetics industry avoidance of animal testing, COX-2, 113, 126 CRD (Colorectal distension) methods, 94–97 procedures, 93–97 quantifiable responses, 94 in the rat, 77 responses in mice, 96 0035_frame_IDX Page 308 Wednesday, May 2, 2001 7:00 AM 308 Cre recombinase, 54, 55–56 Creatine determination in vivo in the brain by MRS, 252 Creatine phosphate determination in vivo in the brain by MRS, 252 CrossRef, Croton oil, 99 Cruelty to experimental subjects, see Ethical and legal issues Culture media for neuron experiments, 158 Cutaneous pain analgesic drug effects, 286 induction in human subjects, 265, 266 pharmacological studies, 286 Cyanine staining, 192 Cyclophosphamide, 102 Cystitis, 94, 102 Cytokine measurements in rodents, 110–126 Cytokines antiinflammatory, 110 detectability, 110 following SNT, 117 inflammatory, 87 in neuropathic pain state maintenance, 113 proinflammatory, 110 protein detection and quantification, 118–122 techniques for study, 114–122 tissue insult response, 110 D DAMGO, 48, 52, 59 DBA/2 mice, 21, 30 Delta-opioid receptor agonists, 101 Delta receptor, 42, 47 coregulation with mu receptor, 52 morphine analgesia in null mutant mice, 50 in null mutant mice, 50 Delta receptor agonists, 52 Delta-selective agonists, 51–52 Deoxygenated hemoglobin (deoxyHb) as MRI contrast agent, 230 Deoxyhemoglobin, 243 Detector patches, 139–140 Dexamethasone, 45 Dextran tracer, 196, 201 fluorescent, 202 Dextromethorphan, 287 effects on heat pain threshold, 286 and temporal summation, 287 DiA, 202 Diabetes formalin test rat response, 73 neuropathic pain, 281 Methods in Pain Research neuropathy models, 83–84 peripheral neuropathy, 79 Dialkylaminostyryl dyes, 202 Diamidino yellow tracer, 202 Digoxigenin reporter, 200 DiI dye, 202 Dimerization of receptors, 52 DiO dye, 202 [11C]Diprenorphine for receptor mapping, 229 Dissectors for neuron counting, 220–221, 223 Distension of hollow visceral structures, 94 duodenum, 97 ileum, 97 jejunum, 97 renal pelvis, 98 urinary bladder, 98–99 urinary bladder stimulation responses, 100 DLPFC (Dorsolateral prefrontal cortex), 255, 256, 257 in back pain, 254 cDNA probes, 200 DNA studies on genetically modified mice, Dogs, 96 Dopamine, 50 receptor binding studied by PET imaging, 227 Dopamine measurement, 136, 136–137, 139 DOP receptor, see Delta receptor DOR receptor, see Delta receptor Dorsolateral prefrontal cortex (DLPFC), see DLPFC (Dorsolateral prefrontal cortex) Doxycycline, 44 DPDPE agonist, 51–52 Drug metabolism affected by genetic differences, 286 Duodenum distension, 97 Dynorphin binding, 42, 50 E E-BioSci, Ecdysone receptor as transactivator, 44 Echoplanar speed imaging (EPI), 231 E coli tetracycline resistance system for transgene expression, 44 Electrical pain induction in human subjects, 266, 272 Electrochemistry assays, 136–137, 139 carbon fiber electrode method, 136–137 with HPLC, 136–137 Electronic publishing and information sources, 7–9 Electrophysiological recording methods, 147–162 recording chambers, 153–154 0035_frame_IDX Page 309 Wednesday, May 2, 2001 7:00 AM Index surgery, 153–154 tissue isolation, 153–154 ELISA (Enzyme-linked immunosorbent assays), 118, 119, 121–122 Embedding medium preparation, 207 Encephalograms for pain assessment, 276–277 Endomorphin binding, 42 beta-Endorphin, 47 beta-Endorphin binding, 42 Endorphin binding, 42 Endorphins in brain and spinal cord, 136 Enkephalin, 51, 53 Binding, 42 in brain and spinal cord, 136 detection by immunocytochemistry, 188 mu receptor binding, 52 Enzyme detecton by immunocytochemistry, 188 Enzyme-linked immunosorbent assay (ELISA), see ELISA (Enzyme-linked immunosorbent assays) Enzymes for neuron isolation, 157–158 EPI (Echoplanar speed imaging), 231 Epi-fluorescence microscopy, 192, 194, 197 Epilepsy, 227, 252 EPON-Araldite embedding medium preparation, 207 Estrogen receptor, 45 Etching embedded immunocytochemistry specimens, 199 Ethical and legal issues, 3–4, 102 avoiding cruelty to laboratory subjects, controlled substance issues, placebos, terminally ill cancer patients, failure to provide adequate pain treatment, withholding of aid to control groups, Evoked potential pain assessment methods in human subjects, 276, 282–283 Exercise-induced muscle pain in human subjects, 271–272 Experimental pain used in human subjects, 281–284 Extracellular recording techniques, 148–151, 153; see also specific techniques nature of pain, 149 schematic, 150 Extracellular sampling techniques, 133–140; see also specific techniques F Fast blue tracer, 202 19F determination in vivo in the brain by MRS, 252 FDG (18-Fluorodeoxyglucose), 226–227 Female reproductive organ stimulation, 102 309 Fentanyl, 48 18F for PET studies, 226–227 Fibromyalgia, 283, 284 Fixation of tissues, 189–190 for electron microscopy, 198 formaldehyde fixative preparation, 203 for light microscopy, 189–191 Flp recombinase, 54 Fluorescein staining, 192 Fluorescence resonance energy transform (FRET) studies, see FRET Fluorescent dyes, 202 Fluorochromes, 192 18-Fluorodeoxyglucose (FDG) in PET imaging, 226–227 Fluorogold tracer, 196, 202 fMRI (Functional magnetic resonance imaging), MRI (Magnetic resonance imaging) Formalin test, 27–29, 111, 112 C-fiber nociceptors, 73 diabetic rat responses, 73 Fos-protein expression, 97 Fractionator for neuron counting, 221 Freeze lesions, 113 FRET (fluorescence resonance energy transfer) studies, Freund's adjuvant, 16, 77 Frozen sectioning, 191 beta-Funaltrexamine, 48, 50 Functional dyspepsia, 284 G GABA determination in vivo in the brain by MRS, 252, 253, 254 light microscopic detection, 191 GABAA detection by immunocytochemistry, 188 GABA detection by immunocytochemistry, 188 GABA measurement, 136 Gabapentin, 28, 70, 73 GADPH, 116 beta-Galactosidase gene as reporter, 46 Galanin measurement in vivo, 137 Genetic correlation of nociceptive assay responses in mouse strains, 15 Genetic differences in drug metabolism, 286 Gerbil, 84 GFAP (Glial fibrillary acidic protein promoter), 112 overexpression, 124 staining, 119 Glial cells activation by cytokines, 113 0035_frame_IDX Page 310 Wednesday, May 2, 2001 7:00 AM 310 cytokine expression for conversion into immunocompetent cells, 126 expression of MHC II, 124 immunocompetent, 123–124 microglia, 123; see also Microglia synthesis of toxins, 126 TNF induction of MHC II expression, 124 Glial fibrillary acidic protein (GFAP) promoter, see GFAP (Glial fibrillary acidic protein promoter) Glucocorticoid receptor, 45 Glucose determination in vivo in the brain by MRS, 252, 253, 255 Glutamate decarboxylase detection by immunocytochemistry, 188 Glutamate detection, 188 Glutamate measurement by detector patch method, 139 determination in vivo in the brain by MRS, 252, 253, 254 by microdialysis, 136 in vivo, 137 Glutamate-receptor antagonists, 101 Glutamine determination in vivo in the brain by MRS, 252 Glutaraldehyde fixative, 198 Glycine detection by immunocytochemistry, 188 by light microscopic methods, 191 H Ham's media, 158 Hargreaves' paw-withdrawal test, see Pawwithdrawal test HCO3- measurement by ISM, 138 1H determination in vivo in the brain by MRS, 252 Heat stimulation withdrawal reflex in human subjects, 278 Heat stimuli, brain regions involved in response, 227–228 Hemodynamic response in fMRI, 231 Heroin, 48 Herpes simplex virus expression systems, 52–53 protein, 44 High Wire Press, High-performance liquid chromatography (HPLC), see HPLC (Highperformance liquid chromatography) Hippocampal neurons, 160 HIV-associated dementia, 110 Methods in Pain Research H+ measurement by ISM, 138 Hollow viscus distension of hollow visceral structures, 76–77 inflammation, 78 Horseradish peroxidase (HRP) staining, 192, 195, 196, 197, 200, 201 Horses, 96 Hot-plate assays, 17, 20–22, 51 animal species behavior variation, 21 increasing-temperature test, 21–22 neural mechanisms, 14 response interpretation, 20–21 standardization of response interpretation, 72–73 HPLC (High-performance liquid chromatography), 136 with electrochemical detection, 136–137 HRP (Horseradish peroxidase) staining, see Horseradish peroxidase (HRP) staining Human cytomegalovirus (CMV) for transgenesis, 44 Human growth hormone expression, 45 Human self-experimentation, 151 Human subjects experimental pain, assessment of, 275–288, 275t pain induction methods, 265–274; see also specific types of pain Hybridization indirect, 200 in situ, 200 Hypalgesia defined, Hyperalgesia defined, 76 Hypertonic saline, 272, 273 for muscle pain induction in human subjects, 279–280, 279–280 Hypoalgesia defined, Hypothalamus, 228 I IASP (International Society for the Study of Pain), guidelines for animal research, 3, 71 Iberiotoxin-sensitive current, 180 IBS (Irritable bowel syndrome), 284 Ibuprofen, 287 ICAM (Cellular adhesion molecules, intracellular), 119, 125, 126 Idiopathic trigeminal neuralgia, 83 IL-10, 110, 111, 117, 122 IL-1α, 117 0035_frame_IDX Page 311 Wednesday, May 2, 2001 7:00 AM Index IL-1β, 110, 111, 112, 113, 117 antibodies, 119 antibodies commercially available, 120 assays, 121 in back pain, 113 ELISA assay, 122 ICAM-1 expression induction, 126 in SNT injury, 122 IL-4, 111, 122 IL-6, 110, 111, 112, 113, 117 antibodies, 119 antibodies commercially available, 120 assays, 121 ELISA assay, 121 in SNT injury, 122 Ileum distension, 97 Immersion fixation, 190 Immersion test for thermal hypersensitivity, 74–75 Immune cell infiltration, 87 Immunocytochemistry definition, 188 electron microscopy and, 197–199 light microscopy and, 188–197 multiple immunolabelling and light microscopy, 193–194 neurotransmitters studied by, 188 staining controls, 195–196 and tract tracing, 196–197 tract tracing, 201 Immunofluorescence staining, 192 protocol for vibratome sections, 204–205 Immunoglobin cellular adhesion molecules, 125 Immunogold staining, 199 post-embedding protocol, 206–207 Immunohistochemistry, 114, 118–119 antibody specificity, 119 with light microscopy, 188–197 Immunolabelling, multiple, 193–194 Immunoperoxidase staining, 192, 199 for electron microscopy, 198 method for electron microscope samples, 205–206 method for vibratome sections for light microscopy, 203–204 Immunostaining methods, 191–194; see also specific methods difficulties, 195 for electron microscopy, 198, 199 for light microscopy, 191–192 Inflammation of a hollow viscus, 78 Inflammatory algogens, 16 Inflammatory pain, see Chronic pain Informatics websites, Inositol complex determination in vivo in the brain by MRS, 252, 254–255 311 In situ hybridization (ISH), 114, 200 difficulties, 114–115 Insula, 246 Integrins, 125 Interferons, 110 Interleukins, 110 International Society for the Study of Pain (IASP), Intracellular Adhesion Molecule (ICAM), see ICAM (Cellular adhesion molecules, intracellular) Intracellular injection methods and immunocytochemistry, 197 Intracellular recording in vitro, 152–153, 154–155 in vivo, 151–152 In vitro extracellular recording techniques, 138–140; see also specific methods schematic, 150 in vivo differences, 139 In vivo extracellular recording techniques; see also In vivo extracellular sampling techniques schematic, 150 In vivo extracellular sampling techniques, 134–138; see also specific methods in vitro differences, 139 Inward-rectifying currents, 179–180 Ion channel measurements, 139–140 Ion channels, 169–182; see also Transducers Ionic currents, 174–179; see also specific ion currents inward-rectifying currents, 179–180 Ion-selective microelectrode (ISM), see ISM (Ionselective microelectrode) IRES (Internal ribosomal entry site), 55–56 Irritable bowel syndrome (IBS), 284 Ischemia, 110 coronary artery occlusion, 103 model of visceral organs, 94 muscle pain, 271–272 role of cellular adhesion molecules, 125 ISM (Ion-selective microelectrode), 137–138, 139 species measured, 138 Itch receptors, 151 J Jejunum distension, 97 K Kappa receptor, 42, 46, 47, 48, 51 agonists, 101 0035_frame_IDX Page 312 Wednesday, May 2, 2001 7:00 AM 312 binding, 49 morphine rewarding properties, 50 null mutant mice, 50, 51 K+ currents, 174–175 characterization, 175 inward-rectifying currents, 179 VGKCs (Voltage-gated K+ currents), 174–175 Ketamine, 287, 288 effects on heat pain threshold, 286 and temporal summation, 287 treatment, 283 Kidney stones, see Ureteral calculosis K+ measurement by ISM, 138 Knockin mice, 55, 56 KOP receptor, see Kappa receptor KOR receptor, see Kappa receptor Kozak sequence, 43 Krox-24 proteins, 102 L LabView software, 245 Lactate determination in vivo in the brain by MRS, 252, 253, 255 Laminin in plating substrate, 159 Lasers for thermal pain induction, 267, 277 Lasers for thermal pain induction in human subjects, 282 Lateral-medullary infarcts, 228 Lectin binding and immunocytochemistry, 197 tracer, 201 Legal consequences, Leukocytes in areas of infection, 125–126 Levo-ascorbic acid, 272 17Li determination in vivo in the brain by MRS, 252 Lidocaine, 84, 98–99, 247 Lipopolysaccharide (LPS)-induced hyperalgesia, 111 LRET (Luminescence resonance energy transfer) studies, LRSC, see Rhodamine staining Luciferase reporter, 44 Luciferin/luciferase ATP assay, 140 Luminescence resonance energy transfer (LRET) studies, Lupus erythematosus, 124 Methods in Pain Research M Magnetic resonance spectroscopy (MRS), see MRS (Magnetic resonance spectroscopy) Magnetoencephalography (MEG), see MEG Major Histocompatibility Complex (MHC), see MHC (Major Histocompatibility Complex) Major Histocompatibility Complex II (MHC II), see MHC II (Major Histocompatibility Complex II) McGill Pain Questionnaire (MPQ), 256, 257, 276 Mechanical allodynia, see Mechano-allodynia Mechanical assays, 23–25; see also specific tests Mechanical pain stimulation in human subjects, 268, 272 Mechano-allodynia, 70, 87, 111 von Frey Fiber Test for, 76 Mechano-hyperalgesia, 75, 86, 87 Mechanosensitivity of neurons, 172 Media for neuron experiments, 158 MEDx software, 245 MEG (Magnetoencephalography), 233–234 MEM (Minimal essential medium), 158 Membrane ion channels, see Ion channels Mexiletine, 84 MHC (Major Histocompatibility Complex), 124–125 MHC genes, 124 MHC I, 124 MHC II (Major Histocompatibility Complex II), 119, 124 Mice; see also Animals; specific strains assays not used in, 16 differences from rats, 13–14, 27 differences from rats in hot-plate assays, 21 differences from rats in thermal assay response, 18 flinch response, 28 genetic correlation of nociceptive assay responses in mouse strains, 15 genetic studies of, 5, 12 increasing use of, 5, 12 knockin mutations, 55, 56 knockout studies in, 12 mutagenesis techniques for opioid system study, see Mutagenesis techniques for murine opioid system study preproenkephalin deficient mutants, 15 SOD mouse, 84 strain variabilities, 29–30, 57–58, 59–60 in hot-plate assays, 21 in tail-withdrawal test, 18 tachykinin-1 mutants, 15 0035_frame_IDX Page 313 Wednesday, May 2, 2001 7:00 AM Index unsuitable for repeated trials, 13 wild-type inbreeding, 58 Microdialysis sampling, 135, 136, 137, 139 Microelectrodes for intracellular recording, 152 Microglia, 123 expression of MHC II, 124 overexpression of GFAP, 124 Microneurography in humans, 150–151 Microspheres, fluorescent, 202 Microtome for sectioning, 191 Mifeprestone (RU486), 45 Migraine, serum levels of Mg+2, 138 Minimal essential medium (MEM), 158 Minnesota Multiphasic Personality Inventory (MMPI), see MMPI MMPI (Minnesota Multiphasic Personality Inventory) in pain measurement, 69 Morphine, 287 analgesia receptor dependence, 50 analgesic action to thermal stimuli, 48 derivatives for acute pain, 242 response in bowel distension, 97 thermal stimulus tests of, 17 visceral pain response, 98 Morphine-6-glucuronide (M6G), 48 Morphometric approaches using MRS, 258 Motor cortex, 228 MRI (Magnetic resonance imaging), 230–233 chronic pain techniques, 242 fMRI, 227, 230–233, 243 acute pain studies, 246 application to clinical pain, 246 back pain studies, 249–251, 254 in clinical setting, 231 correlation with pain perception, 244 data interpretation, 245 experiments, 230–231 measurements, 244 pain intensity studies, 249 SMP studies, 246–248 pain related activations, 235 with PET imaging, 226 phantom limb pain studies, 232–233 mRNA (messenger ribonucleic acid) assays, 114–118 data interpretation, 117–118 mRNA (messenger ribonucleic acid) temporal expression patterns, 122 MRS (Magnetic resonance spectroscopy), 251–258 back pain vs chemical concentrations, 257 data collection and processing, 255 localization in the brain, 254 in vivo, 252–253 Mu agonists, 49–50 313 Mu receptor, 42, 46, 47, 51, 52 agonists, 101 coregulation with delta receptor, 52 immunocytochemical staining, 193 morphine action on thermal stimuli, 48 mutant mice, 48 null mutant mice, 51 spliced forms, 49 Mu receptor agonists, 52 Multiple sclerosis, 110, 124, 125, 252, 282 Muscle pain, 77 analgesic drugs, 288 assessment in human subjects, 278–280 methods, 279 pain and motor control interaction, 280 stimulation in human subjects, 270–271, 271, 272–273 Mustard oil, 16, 78, 99, 102, 287 stimulation in human subjects, 268, 270 Mutagenesis techniques for murine opioid system study, 41–60; see also specific techniques chemical agents, 42 construct design, 57 genes identified and cloned, 42 receptor genes identified and cloned, 42 targeted mutagenesis, 42 targeted mutations, 55, 56 tissue specific mutations, 53–54 transgenesis, 42 viral-based gene constructs, 42 N Na+2 channel blockers, 84 Na+ currents, 176–178, 177 and neuronal excitability, 176 patch electrode measurements, 177 Na+ measurement by ISM, 138 NAA (N-acetyl aspartate) depletion in neurodegenerative diseases, 252 measurement in vivo in the brain by MRS, 252, 254 in back pain, 256 Naloxonazine, 48 Naltrexone, 50 Naltrindole, 48 Naproxen/paracetamol, 288 National Center for Biotechnology Information (NCBI), National Chronic Pain Outreach Association, 243 NCBI (National Center for Biotechnology Information), neo selection cassette, 56, 57 0035_frame_IDX Page 314 Wednesday, May 2, 2001 7:00 AM 314 Nerokinin detection by immunocytochemistry, 188 Nerve growth factor (NGF), 159–160 Nerve inflammation, 87 Neurobiotin labelling, 197 Neurokinin A, 272 measurement in vivo, 137 Neuroma-in-continuity, 81, 82 Neuronal counts, Neuronal culture preparation, 159 Neuronal tracers, 201 Neuronal tract tracing, 200–203 Neuron counting methods, 213–223 Neuron isolation, enzymes for, 157–158 Neuropathic pain, 70, 71, 111 chemotherapy-evoked, 85–87 drug effects on, 73 inflammatory cascade, 122 models, 79–81 from nerve inflammation, 87 postherpetic neuralgia, 84–85 spontaneous vs stimulus-evoked, 81 temporal summation, 281 Neuropeptide Y measurement in vivo, 137 Neurotransmitters studied by immunocytochemistry, 188 NGF (Nerve growth factor), 159–160 NH4+ measurement by ISM, 138 Nitric oxide, 113 measurement, 136–137 synthase inhibitors, 101 NMDA, 73, 268 detection, 196 detection by immunocytochemistry, 188 receptor antagonists, 99 NMDA-antagonist treatment, 283 N-methyl-D-aspartate (NMDA), see NMDA Nociceptive assays; see also Mechanical assays; Thermal assays choice of, 14–16 confounds with increased test sensitivity, 17 dimensions characterizing, 14 thermal, 18–23; see also Thermal assays use of multiples, 15 Nociceptive intensity, 16–18 thermal assays, 17 Nociceptive withdrawal reflex in human subjects, 277–278 NO (nitric oxide), 113 measurement, 136–137 Non-steroidal anti-inflammatory drugs (NSAIDs), see NSAIDs nor-Binaltrophamine (norBNI), 48 Norepinephrine, 80 Norepinephrine measurement, 136, 136–137 Methods in Pain Research NSAIDs (Non-steroidal anti-inflammatory drugs), 28 for acute pain, 242 antinociception in hot-place assays, 21–22 mechanism of action, 17 for muscle pain, 272 response in bowel distension, 97 Null mutant mice, 47–51 basal function changes, 51 receptor interactions, 51–53 tissue specific, 54–55 O Obese sand rat, 84 Occupational injuries primarily back injuries, 250 17O determination in vivo in the brain by MRS, 252 Oligomerization of receptors, 52 Oligonucleotide probes, 200 Opiate receptor, binding studied by PET imaging, 227, 229 κ-opioid agonists, 28 Opioid peptide measurement in vivo, 137 κ-opioid receptor agonists, 99 κ-opioid receptors, 15 Opioid systems, interdependence, 52 Opioids, endogenous, 42–43 OPR-1 receptor, see Delta receptor OPR-2 receptor, see Kappa receptor OPR-3 receptor, see Mu receptor Optical dissector for neuron counting, 220–221, 223 Optical techniques, Orphanin FQ/nociceptin orphan opioid ligand, 17–18 Osmification of electron microscopy samples, 206 OX-42 staining, 119 Oxyhemoglobin, 243 P Paclitaxel, 79, 85–86, 86–87 Pain definition, as essential mechanism, gender differences, 99 hormonal differences, 99 human experiments, induction in, 264–274; see also specific types of pain induction of for human experiments; see also specific types of pain organizations, 0035_frame_IDX Page 315 Wednesday, May 2, 2001 7:00 AM Index PET imaging of, 227–229 psychophysical measurements, 70 purpose of, response similarity to humans, 68–71, 98 as a scientific discipline, as a sensory system, study in human subjects, subjective components, types and stimulated nociceptors, 151 Pain adaptation model, 280 Pain consciousness, 248–249 Parietal cortex, 247 Parkinson's disease, 227 Partial nerve transection (PNT) injury, see PNT Patch-clamp recording in vitro, 156–157 cell-attached patch, 173 and ion channel measurements, 171 pressure clamping, 173 in vivo, 161–162 Paw-flick test, see Paw-withdrawal test Paw-pressure test, 25, 75–76 Paw-withdrawal test, 22, 24, 73–74 interpretation issues, 74 strain sensitivity, 22 PCD (Programmed cell death) studies, 199–200 PCR (Polymerization chain reaction) methods, 31P determination in vivo in the brain by MRS, 252 PECAM (Platelet cellular adhesion molecules), 125, 126 Peltier principle thermodes, 267 Perfusion of excised tissues, 138–139 Perfusion fixation, 189 Periaqueductal gray, 228 Peripheral nerves injury and spontaneous discharge, 151 single unit recording, 151 Peripheral neuropathy, 79, 83 and central neuroinflammatory processes, 124 chemotherapy-evoked, 85–87 immune recognition role, 124 lasers for thermal pain induction, 282–283 models, 81–83 Peripheral sensitization by capsaicin, 270 PET (positron emission tomography), 226–229, 233, 243–244 pain intensity, 249 pain related activations, 235 PET (positron emission tomography) imaging vs fMRI, 227 limitations on radioactive dosing, 227 PHA-L tracer, 201 Phantom limb pain, 79–80 fMRI studies, 232–233 MEG studies, 234 315 Pharmacodynamcs, 285 Pharmacological studies in human subjects, 284–288 important aspects, 285 Phaseolus vulgaris leucoagglutinin tracer, 196, 201 Phosphates measured by MRS, 252 Physical dissector for neuron counting, 220 Phytoestrogen effects on pain responses, 82–83 Pin-prick test, 75 Placebo ethical issues, Plating substrates, 159 PNT (Partial nerve transection) injury, 81–82 confounds, 82–83 Point-resolved spectroscopy (PRESS), see PRESS (Point-resolved spectroscopy) Political consequences of pain research, Poly-d-lysine plating substrate, 159 Poly-L-ornithine plating substrate, 159 Polymerization chain reaction (PCR) methods, see PCR POMC (Proopiomelanocortin) gene, 46, 47 Positron emission tomography (PET), see PET (Positron emission tomography) Post-fixation, 190 Postherpetic neuralgia, 84–85 Potassium chloride, 272 Prefrontal cortex, 228, 246 and back pain, 251 Preproenkephalin deficient mutant mice, 15 PRESS (Point-resolved spectroscopy), 252 Pressure algometry in human subjects, 279 Primates, 99 Prodynorphin gene, 47 Proenkephalin A promoter in mice, 45–46 Profile counts, 214–215, 214–215 Profile ratios, 215 Progesterone receptor, 45 Programmed cell death (PCD) studies, 199–200 Proopiomelanocortin (POMC) gene, 46, 47 Propidium iodide tracer, 202 Prostaglandins, 113 PGE2 as inflammatory mediator, 172 PGe2 inhibition of VGKCs, 175 PGE2 neuron sensitization in vitro, 171 Protein kinase A in nociception, 15, 45 Protein kinase C in nociception, 15 Protocols for microscopy procedures, 203–207 Psychophysical assessment of pain in human subjects, 276, 279 Psychophysical measurements, 70 PubMed, Puffer pipette, 174 Push-pull cannula, 135 Push-pull cannula method, 137 0035_frame_IDX Page 316 Wednesday, May 2, 2001 7:00 AM 316 Methods in Pain Research R S Rabbits, 96 Radiant heat testing tail color effects in, 19 variability, 19 Radiochemicals in PET imaging, 226–227 Radioisotopes decline in use, in PET imaging, 226 replacement by other technologies, Randall-Selitto paw-pressure test, 25, 75–76 Rats; see also Animals differences from mice, 13–14, 27 in hot-plate assays, 21 in thermal assay response, 18 Rats as principal laboratory subject, rCBF (Regional cerebral blood flow) imaging, 227 Receptor binding studies by PET imaging, 227 Receptor interactions, 52 Receptor mapping by PET imaging, 229 Recombinases for conditional targeted mutations, 54–56 cre recombinase, 54 Reference space in histological sections, 216, 219–220 Reflex withdrawal, 12 Remifentanil, 288 Renal pelvis distension, 98 Resiniferatoxin, 101 Restless legs syndrome, 230 Restraints causing SIA, 19, 20 Retinoic acid receptor, 44 Reverse transcriptase polymerase chain reaction (RT-PCR), 114 Rheumatoid arthritis, 124 Rhodamine staining, 192, 197, 203 Ribonuclease protection assay (RPA), see RPA (Ribonuclease protection assay) RNA isolation kit comparison, 115 RNA isolation procedures, 115 RNA probes, 200 RPA (Ribonuclease protection assay), 114 analysis of cytokine mRNA, 116 limitations, 117 pH sensitivity, 116 study of mRNA nodulation, 116–117 RT-PCR (Reverse transcriptase-polymerase chain reaction), 117 RU486 (mifeprestone), 45, 55 S1 (Primary somatosensory cortex), 227–228, 232 MEG studies, 234 pain related activations, 235 S2 (Secondary somatosensory cortex), 227–228, 232 MEG studies, 234 pain related activations, 235 Sciatic nerve, 111 Sectioning for electron microscopy, 198 for light microscopy, 190–191 Selectins, 125 Self-mutilating behavior, 80–81 Sensitization, see Central sensitization Sensory neurons, ion channels in, 172 Serial reconstructions, 215 Serotonin measurement, 136, 136–137 Serum for culture media, 158–159 Serum substitutes for culture media, 158–159 Shingles, 84–85 Shrinkage in sample preparation for neuron counting, 221 SIA (Stress-induced antinociception), 17 in abdominal constriction test, 26–27 animal strain dependence, 18 confounds, 17 endogenous opioid peptides involved, 60 in formalin test, 28 highly noxious stimuli to reduce impact, 18 in hot-plate assays, 21 injection-related, 17–18 in laboratory animals, 13 mice lacking enkephalin, 51 in null mutant mice, 47–48 in radiant heat testing, 19 restraint effects, 19, 20 Simulated echo-acquisition mode (STEAM) spectroscopy, see STEAM (Simulated echo-acquisition mode) spectroscopy Single nucleotide polymorphisms (SNPs) in mice, Single photon emission computerized tomography (SPECT) imaging, see SPECT (Single photon emission computerized tomography) imaging Skin burn model, 270, 287 Skin nociceptors, 267 Small bowel distension, 97 SMP (Sympathetically mediated pain), fMRI studies of, 246–248 SNPs (Single nucleotid polymorphisms) in mice, 0035_frame_IDX Page 317 Wednesday, May 2, 2001 7:00 AM Index SNT (Spinal nerve transection) injury, 82, 113, 114 assays, 122 cytokine assays, 121 mRNA studies, 116–117 SOD mouse, 84 Somatosensory cortex primary (S1), see S1 secondary (S2), see S2 Somatostatin measurement in vivo, 137 Southern blot analysis, 44 Soy effects on pain responses, 82–83 Species used in pain research, 12 SPECT (Single photon emission computerized tomography) imaging, 229–230, 233 pain related activations, 235 Spinal cord fluid sampling by push-pull cannula, 135 Spinal cord injury pain, 230 Spinal nerve transection (SNT) injury, see SNT (Spinal nerve transection) injury Spinal superfusion method, 135 SPM (Statistical parametric mapping) program, 227 SQUID (Superconducting quantum interference device) magnetometers, 233 Staining; see also specific stains fluorescent dyes, 202 Statistical parametric mapping (SPM) program, 227 STEAM (Simulated echo-acquisition mode) spectroscopy, 252 data collection and processing, 255 Stereologic methods, 215, 217 models, 215, 217 procedure, 221–222 protocols, 218–223 Stereologic neuron counting methods, 213–223 Steroid hormone binding domain methods, 45 Stimulus control and standardization, Stimulus intensity effects, 16–17, 16–18 Stimulus modality classification, 14 Strain dependence of mouse responses, 30, 44, 57–58, 59–60 Streptavidin detection, 200 Streptozocin, 83–84 Stress-induced antinociception (SIA), see SIA Stroke, 252 Substance P, 272 in brain and spinal cord, 136 detection, 194 detection by immunocytochemistry, 188 measurement in vivo, 137 nociceptive, 112 Suction electrodes, 149 Summation of muscle pain, 273–274 317 Summation of nociceptive activity in human subjects, 268–269 Supraspinally integrated behaviors, 73 Sympathetically mediated pain (SMP), see SMP (Sympathetically mediated pain) Syringomyelia, 282 T Tachykinin-1 mutant mice, 15 differences from wild-type, 16–17 Tail-clip test, 24–25 Tail-flick test, 51, 72, see Tail-withdrawal test tail color effects, 72 Tail-immersion test, see Tail-withdrawal test Tail-withdrawal test, 18–20, 72 avoidance learning, 23 radiant heat testing variability, 19 SIA due to retraint, 19 tail color effects in radiant heating test, 19 Targeted mutations, 56, 57 Taxol, 85 TechDox, Temporal summation of nociceptive activity in human subjects, 268, 278 Temporomandibular joint sampling by push-pull cannula, 135 Tetracaine, 101 Tetracycline-controlled system, 55 Thalamus, 228, 232, 257 in back pain, 254 in central pain, 230 measurement of noxious stimulus impact, 149 pain related activations, 235 Thalidomide, 111 Theophylline, 288 Thermal assays, 75; see also specific tests good predictors of analgesic potential, 18 paw-withdrawal assay, 22 spinal vs supraspinal, 22–23 Thermal hypersensitivity immersion test, 74–75 Thermal pain induction in human subjects, 267 Tic douloureux, 83 Tissue preparation for immunocytochemistry, 189–190 Tissue sectioning, 190–191 from embedded blocks, 191 (TNBS) Trinitrobenzene sulfonic acid, 96 TNF (Tumor necrosis factor), 87, 110–113, 117 antibodies, 119 antibodies commercially available, 120 regulation of ICAM, 126 Tourniquet model, 271, 288 Tract tracing, 200–203 Tract tracing and immunocytochemistry, 196–197 0035_frame_IDX Page 318 Wednesday, May 2, 2001 7:00 AM 318 Transducers characterization, 172 chemical, 173–174 mechanical, 173 properties in nociceptor processing, 172 thermal, 173 Transgenesis, 43–46 definition, 43 dexamethasone, 45 ecdysone receptor, 44 E coli tetracycline resistance system for transgene expression, 44 estrogen receptor, 45 glucocorticoid receptor, 45 herpes simplex virus protein, 44 human cytomegalovirus (CMV) used in, 44 human growth hormone expression, 45 irreducible transgenes, 43, 44–45 Kozak sequence, 43 luciferase reporter, 44 mice for opoid system experiments, 45–46 for opioid system studies, 45–46 progesterone receptor, 45 retinoic acid receptor, 44 targeted mutations, 46–53 Transposons, 57 Trigeminal ganglia, 136, 157–158 Trigeminal neuralgia, 83 Trinitrobenzene sulfonic acid (TNBS), 96 Trinitrophenol stimulant, 173 Trophic factors in plating media, 159–160 Tumor necrosis factor (TNF), see TNF (Tumor necrosis factor) TUNEL (TdT-mediated dUTP-biotin nick end labelling) method, 188, 199–200 Turpentine, 78, 96, 99, 101 Tursky electrode, 266 Methods in Pain Research VGCCs (Voltage gated calcium currents), see Ca+2 currents VGKCs (Voltage-gated K+ currents), see K+ currents VGSCs (Voltage-gated sodium currents), see Na+ currents Vibratome sectioning, 190 Vincristine, 79, 85–86 Viral vectors for transgenesis, 52–53 Visceral pain animal models lacking, 93 colonic-rectal distension model, 94–97 distension of hollow visceral structures, 94 distension of hollow viscus, 76–77 emulation by abdominal constriction test, 27 induction in human subjects, 274, 274 κ-opioid receptors in, 71 methods, 6, 93–104, 94–97; see also specific methods models, 95 various nature, 103 Visual analog scales (VAS), see VAS Voltage gated calcium currents (VGCCs), see Ca+2 currents Voltage gated sodium currents (VGSCs), see Na+ currents von Frey Fiber Test, 23–24, 70, 76, 286 W Web of Science citation indices, Websites of interest, 7–9 Wheat-germ agglutinin (WGA-HRP) tracer, 201 Whiplash-injury patients, 283 Wick method, 134 Wind-up, 149, 268 Withdrawal reflex in human subjects, 277–278 Writhing test, see Abdominal constriction (writhing) test U UBD (Urinary bladder distension), 98–99 Ureteral calculosis, 78–79, 94, 97–98 Urinary bladder distension (UBD), 98–99 Urinary bladder irritants, 99–102, 100 Urinary bladder stimulation responses, 100 X Xenon-133 for CBF mapping, 229 Xylene, 99, 101 Y V Varicella-zoster virus, 84–85 VAS (Visual analog scales), 276, 278–279 VDS (Verbal descriptor scales), 276 Verbal descriptor scales (VDS), 276 Vertex potentials, 276–277 Yeast shuttle vector, 57 induction of for human experiments, 264–274 Z Zymosan, 16, 96, 111 ... disciplines 1.2 DEFINING PAIN The definition of pain has been elusive for a variety of reasons deriving from the idiosyncratic history of pain research, 3,4 but principally from the failure to distinguish... AM Methods in Pain Research Recognition of pain as a scientific discipline is relatively recent, deriving largely from the formation of pain clinics and large-scale organizations, i.e., the International... demonstrated to be painful when injected into humans, and produce nociceptive behaviors in animals when injected into the skin (e.g., capsaicin, bradykinin, prostaglandins, formalin) or peritoneal