Concluding Remarks

It is a great pleasure for me to write the concluding remarks for the book Analgesia, Methods in Molecular Biology Series, edited by Arpad Szallasi. Arpad Szallasi is very well known in the research field, especially with his descrip- tion of the vaniloid receptor VR1 or now better known as TRPV1 receptor, which may be a major novel target for drugs. Clinicians are not very familiar with (or interested in) basic research in pain medicine. However, the infrastructure of basic knowledge in pain medicine begins with research and fundementals of biology. The problem for clinicians also may arise from how basic knowledge and research of pain medicine is presented to the readers. I am very happy to see in this book that all topics have been presented and they have a systematic approach for the readers. The book opens with experimental pain models (Chapters 1–10) which is also very important to conduct studies even for clinical purpose. This section gives detailed information about animal models for acute, chronic, inflammatory, neuropathic, orofacial pain, and migraine. The second step (Chapters 11–15) deals with human pain models. The chapters (Chapters 17–30) on genetic determinants of pain sensitivity and gene expression changes during pain, which is a new and interesting area in pain medicine, may be very influential in the near future. These chapters also give insights into drug discovery and development. Methods for the study of mechanism of action of anesthetic (Chapters 1 and 31) and analgesic drugs (Chapters 16–19) are also presented. The chapter (Chapter 33) on “Proteomics and metabolomics and their application to analgesia research” is new to us. Future therapies are described including gene-based approaches (Chapter 22) and stem cell therapy (Chapter 25) which may be speculative for today but has a future. Last, methods are described for clinical studies in (Chapters 35–38) which also have important input for clinicans. This book is a comprehensive coverage of research methods in analgesia from target discovery through target validation and drug development to preclinical trials, and from in silico methods and cell lines through drosophilia and mice to patients. I congragulate Arpad Szallasi and all contributing authors for their great efforts for this book. I do believe it is a great contribution to pain medicine.

Serdar Erdine, MD, FIPP President World Institute of Pain

555 In d e x

A induced by ...... 48 intradental perfusion in rats...... 95 Acceptor photobleaching...... 201–203 intradermal injection in humans...... 170 Acetaminophen...... 166, 181, 182, 189, 541 receptor TRPV1...... 223, 237, 425 Acetic acid-induced writhing test...... 370 Carbachol...... 194, 197–199, 203 Acetone pain model...... 45, 47–48 Carbamazepine...... 541–543 Acid-depressed feeding (mice)...... 81–85 Carrageenan...... 44–46, 50, 65 Acid-depressed intracranial self-stimulation Cathecol-O-methyltransferase (COMT) activity, (rats)...... 82–83 enzymatic assay...... 379 Acute surgical pain...... 31–38 Cathecol-O-methyltransferase (COMT) gene, human a2-Adrenergic agonists...... 545 cDNA constructs...... 377–379, Allen Brain bank...... 310, 315 381–382, 384–386 ...... 33, 34, 37, 41–43, generation of haplotypic variants...... 377–391 47, 49, 53, 67, 69, 95, 98, 155, 170, 173, 327, 328, real-time PCR to determine 335, 338, 339, 342, 475, 548, 549 transcripts...... 380–381, 388–391 Amplicon...... 283, 290–293 Cathecol-O-methyltransferase (COMT) Atropine...... 83, 194, 197–199 polymorphism...... 375–392, 535 Axopatch, electrophysiology...... 224 Central sensitization...... 31, 33, 42, 43, 170, 476 B Channelopathies...... 310 Channel open probability...... 230–233, 235 Bidirectional sequencing...... 311, 316–318 CHO cells Bioinformatics...... 310–311, 313–316, 416, 527 stably expressing M2/M3 receptors...... 193, 195 Bispectral EEG analysis...... 498 Chronic constriction injury (CCI) model Bone marrow stromal cell (rat)...... 327–335 infraorbital nerve...... 95–97, 99–100 Bone metabolism modulators...... 545–546 sciatic nerve...... 44, 157 Botulinum toxin...... 548–549 Chronic pain...... 42, 43, 80, 155–157, 297, Brain receptor imaging...... 529–531 298, 506, 508, 509, 511, 540, 541, 545, 548, 550 C Clinical correlates of experimental pain...... 156–157 Clinical data collection Caenorhabditis elegans ethical considerations...... 404 behavioral assays...... 8–10 prospective study...... 405–406 chemotaxis assay...... 9 retrospective study...... 406–407 immobility assay...... 8 Clustering of genetic traits...... 266, 267 mating efficiency assay...... 9–10 Cold pressor model...... 165–167 mutant rescue assay...... 14 Collagen-induced arthritis...... 70 mutant screening and selection...... 13–14 Colon of the rat...... 119–121, 134–135 Calcitonin gene-related peptide (CGRP) Complete Freund’s adjuvant (CFA)...... 44–47, RIA for CGRP-like immunoreactivity...... 109 51, 65, 69–77, 94–96, 98, 101 Calcium imaging in single cells...... 248–249 Conditioned place preference test...... 364 Calcium microfluorometry...... 239, 241 Congenital inability to experience pain...... 309, 310 Cannabinoids...... 239, 547 Cortical spreading depression...... 106, 108, 111–112 Capnography...... 496, 497, 501 Cyclic AMP (cAMP) assay Capsaicin in forskolin-stimulated cells...... 368, 423 acute pain induced by capsaicin...... 45, 48 in PGE2-stimulated cells...... 423, 425

557 An a l g e s i a 558 Index

D Glyceryl trinitrate...... 107, 108 G-protein coupled receptors (GPRC)...... 239, 531, Dental impaction pain...... 175–189 539, 546, 547 Dental injury model...... 94–99 Grip force test procedure...... 46 Determining anesthetic concentration...... 4–5, 10–12 Dorsal root ganglion (DRG) neuron H culture of...... 240–241, 245 Halothane...... 4, 8, 11, 13, 15, 16, Drosophila melanogaster 64, 129, 147, 437–439, 441 mechanical assay...... 20 Hardy-Weinberg equilibrium (HWE)...... 400, 414 mutagenesis...... 20, 21 Hargreaves method...... 43, 46, 50 thermal nociception assay...... 24–25 HEK293 cells...... 311, 312, 319–321, 425 transposable elements...... 21 Heritability of pain traits dsRNA-containing P-element vectors...... 23 broad-sense...... 262 Dynamic FRET assay narrow-sense...... 262, 263 imaging GPCRs by...... 198–201 Herpes simplex virus (HSV), gene and E RNAi delivery...... 351 Hind-paw pressure test...... 370 Erythema formation scale...... 162, 163 Human mesenchymal stem cells...... 337–345 Erythromelalgia...... 309 Human pain models...... 157, 518 Esophagus of the ferret/mouse...... 122–124, 137–138 Human Protein Atlas...... 310, 315 Evan’s blue dye extravasation assay...... 106, 108, 110 ...... 33, 34, 37, 41–43, 46, 47, F 49, 50, 52–54, 57–65, 67, 68, 95, 98, 155, 159, 160, 162, 163, 169–173, 239, 275, 338, 339, 341–342, First perceptible relief...... 180 422, 475, 480, 481, 540, 545, 546, 548, 550 Fisher 344 rats...... 263 I Fixed-temperature water bath...... 46, 50–51 Fluo-3, intracellular Ca2+ measurement...... 428–429 IMT504...... 328, 329, 332, 335 Fluorescence microscopy...... 342–343 Inbred mouse strains Fluorogold...... 95, 97 recombinant...... 263–264 Formalin pain model...... 44, 45, 47 standard...... 263–264, 273 Free radical scavengers...... 551 Increasing temperature hop plate...... 58, 59, 61, 63–65 Functional magnetic resonance imaging Increasing temperature water bath...... 58, 59, 61, 63–65 (fMRI)...... 520, 522–523, 530 Inflammatory pain...... 44, 67–77 Inhalational anesthetics...... 437–443 G In-silico haplotype mapping...... 273 Gabapentinoids...... 542–543 Ion channels...... 23, 93, 202, Gallbladder of opossum...... 141–144 209–220, 223–236, 239, 247, 255, 256, 445, 549 Gastrocnemius incision pain...... 34–36 L Gel electrophoresis, two-dimensional...... 458, 459 Gene Expression Omnibus...... 310, 315 Laparotomy pain...... 36 Gene knock-down by small interfering RNA Laser Doppler flow meter interference...... 23, 261 for measuring UV-B-induced Genetic correlation, calculation of...... 264 ...... 160 Genomic DNA for meningeal blood flow...... 108, 110–111 preparation of...... 397, 407, 412 Lentiviral vectors...... 298, 299, 302, 304, 306 purification of...... 408–411 Lewis rats...... 70 Genotype-phenotype association...... 403 Likert scale...... 184, 488, 490 Genotyping Linkage analysis of inherited pain conditions...... 309–324 collection of human oral Lipofectamin, transfection by...... 193, 196, 216, mucosa for...... 396, 405, 407, 408 220, 224, 226, 312, 320 human blood...... 405, 406 Lipoparticles...... 446–455 mouse...... 365 Liquid chromatography-mass spectrometry TaqMan SNP genotyping assay...... 398, 411 (LCMS)...... 457, 458, 462, 470–471 GH4C1 cells...... 211, 212, 216, 219 Live cell imaging...... 191–206 An a l g e s i a 559 Index M Optimem...... 193, 196, 212 Orofacial pain...... 93–102 Meaningful relief...... 180, 181, 185, 189 Osteoarthritic (OA) pain...... 44, 546 Mechanical allodynia...... 47, 67, 69, 95, Osteotomy pain...... 33, 37 98, 335, 338, 339, 342, 549 Meningeal blood flow...... 108, 110–111 P Meninges...... 106, 110 Pain assessment Metabolomics...... 457–472 Likert scale in...... 184, 488, 490 Migraine models standardizing...... 483–491 animals...... 106, 110, 111 variability in...... 484–485, 487, 550 human...... 106, 108, 109, 112 Pain-depressed behavior...... 79–90 Minimum erythema dose (MED)...... 160–163 Pain genetics...... 275 Motor coordination behavior Pain imaging (Rotarod apparatus)...... 339 functional...... 518, 519, 521–526, 528, 534 Multidimensional scaling (MDS)...... 266, 267 metabolic...... 519, 526 Multilineage progenitor cell...... 424, 432 molecular...... 519, 521–528, 535 Multiple Primer Extension Pancreas of the rat...... 124–125, 138–141 (MPEX)...... 398–400, 411–413 Paroxysmal extreme ...... 309 Mustard oil...... 94–96, 98, 101 PCR-RFLP (restriction fragment length N polymorphism)...... 397–398, 409 Peak pain intensity difference (PPID)...... 185 nAChRa7 coding sequence...... 213 Peak pain relief...... 185 ND7/23 cells...... 311, 312, 319–321, 323 Peltier device...... 162, 171 Nerve growth factor (NGF)...... 240, 245, 423, Peripheral sensitization...... 33, 42 427, 428, 548 Personalized medicine...... 396 Neuropeptide release measurement...... 249–251 Photolabeling...... 437–443 Neuropeptides...... 95, 101, 106–109, Picospitzer apparatus...... 224 239, 249–251, 257 Pin prick test procedure...... 46, 52 Nitric oxide (NO) release, measurement of...... 424 Plantar incision pain...... 34, 35 NMDA antagonists...... 170, 547–548, 550 Plantar test apparatus, Ugo Basile...... 339 Nociception...... 19–27, 34, 35, 41–55, Plasmids 57, 58, 101, 170, 266, 267, 280, 310, 350, 370, 550 expression vector pCDNA3.1...... 193 Non-invasive transcranial direct current linearization...... 216 stimulation...... 505–512 Miniprep kit to obtain plasmid DNA...... 215 Non-stationary noise analysis...... 228–230 plasmid 8.91...... 298 Non-steroid anti-inflammatory drugs plasmid MD-G...... 298 (NSAIDs)...... 68, 105, 106, 166, pTrip-CMV-WPRE...... 298, 302 178, 179, 182, 185, 186, 478, 540–541 Polymerase chain reaction (PCR) Noxious heat threshold...... 57–65 allele-specific PCR (AS-PCR)...... 398, 399, 410, 411 O reaction mix...... 214, 288 real-time quantitative PCR...... 283 Oligonucleotides...... 261, 290, 317–319, RT-PCR...... 213, 279–294, 430 327–335, 397–399, 411–412, 417 Positron emission tomography (PET)...... 508, 520, m-Opioidergic neurotransmission, 522–526, 529, 530, 532, 535 imaging of...... 531–534 Potassium channel openers...... 549 Opioid partial agonism...... 363–373 Preemptive analgesia...... 475–481 [3H]Opioid receptor binding assay...... 364 pRES-green fluorescent protein (GFP)...... 224, 226 m-Opioid receptor gene (OPRM1) Preventive analgesia...... 476, 477 isoforms, MOR3...... 421–433 Principal component analysis (PCA)...... 266, 267, 467, 468 Opioids...... 34, 35, 58, 81, 159, 160, Procedural sedation...... 493–502 166, 170, 177, 185–187, 191, 275, 363–373, Proteomics, label-free quantitative...... 460, 463 395–417, 421–433, 478, 479, 481, 501, 512, 525, ProteOn optical biosensor chip...... 447, 451 526, 529, 531–535, 539–540, 542, 545, 548, 550 Pulse oximetry...... 495, 497, 498, 501 Optical biosensors...... 445–455 Purification of viral particles...... 299 An a l g e s i a 560 Index R T

Radiant heat assay Tail-flick assay...... 54, 364, 365, 369–372 (Hargreaves method)...... 43, 46, 50 Tail incision pain...... 36, 37 Randall-Selitto paw pressure apparatus...... 68 TaqMan SNP genotyping...... 398, 411 Reduction of spontaneous activity by adjuvant T4 DNA ligase...... 211 (RSAA) model...... 69, 75–77 Temporomandibular joint disorder...... 375–392 Regenerative medicine...... 338 Testis of the dog...... 129, 146–147 Resiniferatoxin...... 58, 59, 280 Thoracotomy pain...... 34, 36 Resriction enzymes Transdermal lidocaine...... 544 BamHI...... 211, 214 Transient transfection XhoI...... 211, 214 Clonfectin kit...... 424, 430 RNA interference (RNAi)...... 2, 23, 347–360 in HEK cells...... 226 RNA isolation...... 280, 292 Tricyclic antidepressants (TCAs)...... 170, 542, 544 RNA quantification...... 280 Triptans...... 105, 106, 239, 544 RNeasy mini kit...... 280, 285, 286 TRIzol reagent...... 285–286 Rotarod apparatus...... 339 TRPV1 receptors, in HEK cells...... 226–230 RT-PCR, gel-based...... 279–294 U S Ultraviolet light (UV-B) pain model...... 159–163 Saalman multitester...... 160, 161 Ureter of the guinea pig...... 127–129, 145–146 Sciatic nerve injection with RNAi...... 353–356 Urinary bladder of the mouse...... 126–127, 144–145 SDS-PAGE...... 353–354, 357, 464 Uterine cervix of the rat...... 129–130, 147–148 Selective breeding of mice...... 262–263 Single fiber recording...... 120, 244, 251 V Single nucleotide polymorphism (SNP)...... 269, Visceral pain...... 34, 35, 115–153 274, 316, 382, 398, 401, 402, 408–411, 414–416, 535 Visual analog pain score...... 184 Single photon emission computed tomography Volatile anesthetic...... 1–16, 108, 439 (SPECT)...... 520, 523–526, 529 von Frey filaments...... 98, 334, 335, 480 Site-directed mutagenesis...... 311, 318–319, 378, 383 Skin-nerve preparation...... 238, 240, 244 W Small bowel of the rat or mouse...... 117–119, 132–134 Western blotting...... 354, 355, Small interfering RNAs (siRNAs)...... 23, 347, 348, 350 357–359, 379–380, 387–388, 426 Sodium channel Na 1.7 gene v Whole blood Total RNA Kit™...... 280, 287 SCN9A...... 309 Whole cell patch-clamp...... 225, 312 Spared nerve injury...... 157, 339, 341 Whole genome amplification (WGA)...... 275, 407, 408 Spontaneous locomotor activity...... 34, 76 Whole genome scan...... 310, 314 Stem cell transplantation...... 337–345 Stomach of the rat...... 121–122, 135–137, 151 Z Subcostal incision pain...... 36, 37 (SP)...... 106, 109, 110, 540, 549 Ziconotide...... 544–545