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5‐Hydroxytryptamine Receptor Assays

Katherine W. Figueroa1,  Graeme R. Martin2,  M. Teresa Pulido‐Rios3

1Johnson & Johnson PRD, La Jolla, California
2Takeda Research Investment, Palo Alto, California
3Theravance, South San Francisco, California



Publication Name: 
Current Protocols in Pharmacology
Unit Number: 
Unit 4.19
DOI: 
10.1002/0471141755.ph0419s46
Online Posting Date: 
September, 2009
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Abstract

5-Hydroxytryptamine (5-HT) receptors, by virtue of their broad expression pattern in peripheral and central tissues, regulate diverse physiological and behavioral responses through the activation of fourteen molecularly distinct receptor subtypes. The tissue-specific distribution of these receptors confers specificity for the actions of serotonin and highlights the therapeutic potential of serotonin receptor modulators. To better assess this therapeutic potential, it is useful to characterize serotonergic agonists and antagonists in physiologically relevant organ systems. Provided in this unit are twelve tissue bath assays using vascular and smooth muscle tissues isolated from guinea-pig, rat, and rabbit. These tests make possible the analyses of compounds at nine serotonin receptor subtypes. Curr. Protoc. Pharmacol. 46:4.19.1-4.19.45. © 2009 by John Wiley & Sons, Inc.

Keywords: serotonin; 5-hydroxytriptamine; 5-HT

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Inhibition of the 5-HT1A Receptor–Mediated Twitch Response in Guinea Pig Transmurally Stimulated Ileum
  • Basic Protocol 2: 5-HT1B Receptor–Mediated Contraction of Rat Caudal Artery
  • 5-HT1D Receptors
  • Basic Protocol 3: 5-Hydroxytryptamine 1D Receptor Assays
  • Alternate Protocol 1: 5-HT1D Receptor–Mediated Contraction of Rabbit Cerebral Arteries
  • Basic Protocol 4: 5-HT2A Receptor–Mediated Contraction of Rabbit Thoracic Aorta
  • Basic Protocol 5: 5-HT2B Receptor–Mediated Contraction of the Rat Stomach Fundus
  • Basic Protocol 6: 5-HT2C Receptor–Mediated Contraction of the Rat Jugular Vein
  • Basic Protocol 7: 5-HT3 Receptor–Mediated Contraction of Guinea Pig Ileal Smooth Muscle
  • 5-HT4 Receptors
  • Basic Protocol 8: 5-HT4 Receptor–Mediated Relaxation of Rat Esophageal Tunica Muscularis Mucosa
  • Alternate Protocol 2: 5-HT4 Receptor–Mediated Contraction of Guinea Pig Distal Colon
  • Basic Protocol 9: Post-Junctional 5-HT7 Receptor–Mediated Relaxation of Guinea Pig Isolated Ileum
  • Alternate Protocol 3: 5-HT7 Receptor–Mediated Relaxation of the Endothelium-Denuded, Precontracted Rabbit Jugular Vein
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Inhibition of the 5-HT1A Receptor–Mediated Twitch Response in Guinea Pig Transmurally Stimulated Ileum

 Materials
  • 200- to 450-g male Hartley guinea pig (Charles River Laboratories)
  • CO2 (e.g., gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 32° ± 1°C
  • Standard 5-HT receptor antagonists (see recipe and steps 6 and 12)
  • 10 mM pargyline (see recipe)
  • 1 mM phenoxybenzamine hydrochloride (see recipe)
  • Standard 5-HT agonists (e.g., 5-CT, 5-HT; see recipes)
  • Test compound(s)
  • Euthanasia chamber (Harvard Apparatus)
  • Surgical instruments: scalpel, scissors
  • Petri dishes
  • 10-ml syringes
  • 1.0 surgical silk
  • Fixed tissue holder fitted with two parallel, bipolar, 2 × 0.06–cm platinum electrodes (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue suspension wire
  • 10-ml tissue bath (also see unit 4.4) including:
    • Electrical stimulator (e.g., Grass S88 plus Grass stimulator isolation units or Digitimer D330)
    • Thermostatically regulated water circulator (WPI)
  • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)

Basic Protocol 2: 5-HT1B Receptor–Mediated Contraction of Rat Caudal Artery

 Materials
  • 250- to 350-g male Wistar rats (Charles River Laboratories)
  • CO2 (e.g., gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 32° ± 1°C
  • 10 mM pargyline (see recipe)
  • 1 mM phenoxybenzamine (see recipe)
  • 1 mM U-46619 (see recipe)
  • Standard 5-HT receptor antagonists (e.g., cyanopindolol; see recipe and step 10)
  • Standard 5-HT receptor agonists (e.g., 1 µM 5-CT or 5-HT; see recipes)
  • Test compound(s)
  • Euthanasia chamber (Harvard Apparatus)
  • Dissecting instrument: scalpel, scissors
  • Petri dishes
  • Fixed tissue holder with stainless steel wire mount (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue suspension wire
  • 10-ml tissue bath (also see unit 4.4) including (WPI):
    • Thermostatically regulated water circulator
    • Polygraph/chart recorder/electronicdata acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)
Basic Protocol 3:  5-Hydroxytryptamine 1D Receptor Assays
 Materials
  • 200- to 450-g male Dunkin-Hartley guinea pigs (Charles River Laboratories)
  • CO2 (gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 32° ± 1°C
  • Standard 5-HT2 receptor antagonist (e.g., methiothepin; see recipe)
  • 10 mM pargyline (see recipe)
  • 1 mM phenoxybenzamine (see recipe)
  • 1 mM U-46619 (see recipe)
  • Standard 5-HT receptor agonist (e.g., 1 µM sumatriptan; see recipe)
  • Test compound(s)
  • Euthanasia chamber (Harvard Apparatus)
  • Dissection instruments: scalpel, scissors
  • Petri dishes
  • Fixed tissue holder with stainless steel wire mount (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue suspension wire
  • 10-ml tissue bath equipment (WPI)(also see unit 4.4) including:
    • Thermostatically regulated water circulator (WPI)
    • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)
Alternate Protocol 1:  5-HT1D Receptor–Mediated Contraction of Rabbit Cerebral Arteries
 Additional Materials (also see Basic Protocol 3)
  • 2.5- to 3.5-kg male New Zealand white rabbits (Charles River Laboratories)
  • Pentobarbitone/sodium heparin (Henry Schein)
  • 1 mM indomethacin (see recipe)
  • 4 M KCl
  • 10 mM acetylcholine (see recipe)
  • Small animal guillotine (Harvard Apparatus)
  • Binocular microscope
  • 0.04 × ~3–in. long stainless steel wire roughened by rubbing with steel wool

Basic Protocol 4: 5-HT2A Receptor–Mediated Contraction of Rabbit Thoracic Aorta

 Materials
  • 2.5- to 3.0-kg male New Zealand white rabbits (Froxfield, Hampshire, UK)
  • Pentobarbitone/sodium heparin (Henry Schein)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 32° ± 1°C
  • 10 mM pargyline (see recipe)
  • 1 mM benextramine tetrahydrochloride (see recipe)
  • Standard 5-HT agonists (i.e., 5-HT; see recipe)
  • Standard 5-HT2A receptor antagonists (agonist-dependent, e.g., Ketanserin; see recipe and step 8)
  • Test compound(s)
  • Surgical instruments
  • Petri dishes
  • 0.04 × ~3–in. long stainless steel wire roughened by rubbing with steel wool
  • Fixed tissue holder with stainless steel wire mount (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue-suspension wire
  • 10-ml tissue bath (WPI) (also see unit 4.4) including:
    • Thermostatically regulated water circulator (WPI)
    • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)

Basic Protocol 5: 5-HT2B Receptor–Mediated Contraction of the Rat Stomach Fundus

 Materials
  • 250- to 350-g male Sprague Dawley rats (Charles River Laboratories)
  • CO2 (gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Tyrode's solution with and without corticosterone, cocaine, and ketanserin (see recipe), 37° ± 1°C
  • Standard 5-HT antagonists (e.g., SB204741; see recipe and step 9)
  • 10 mM pargyline (see recipe)
  • 10 mM benextramine tetrahydrochloride (see recipe)
  • Standard 5-HT receptor agonist (e.g., 5-HT, -methyl-5-HT, 5-CT; see recipe)
  • Test compound(s)
  • 4 M KCl
  • Airtight euthanasia chamber (Harvard Apparatus)
  • 23-G hypodermic needle and 5-ml syringe
  • Surgical instruments: scalpel, scissors, fine scissors with sharp pointed edges, curved forceps
  • Petri dishes
  • Blade (razor blade)
  • 1.0 surgical silk threads
  • Fixed tissue holder with stainless steel wire mount (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue suspension wire
  • 10-ml tissue bath (WPI) (also see unit 4.4) including:
    • Thermostatically regulated water circulator (WPI)
    • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)

Basic Protocol 6: 5-HT2C Receptor–Mediated Contraction of the Rat Jugular Vein

 Materials
  • 250- to 350-g male Sprague-Dawley rats (Charles River Laboratories)
  • CO2 (gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 37° ± 1°C
  • 10 µM ketanserin (see recipe)
  • 10 mM pargyline (see recipe)
  • 1 mM phenoxybenzamine (see recipe)
  • Test compound(s)
  • Standard 5-HT agonist (see recipe)
  • Standard 5-HT antagonists (e.g., RS102221 or mesulergine; see recipe and step 9)
  • Euthanasia chamber (Harvard Apparatus)
  • Surgical instruments: scalpel, scissors
  • Polypropylene tubing (1-mm o.d.)
  • Petri dishes
  • Fixed tissue holder with stainless steel wire mount (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue-suspension wire
  • 10-ml tissue bath (WPI) (also see unit 4.4) including:
    • Thermostatically regulated water circulator (WPI)
    • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)

Basic Protocol 7: 5-HT3 Receptor–Mediated Contraction of Guinea Pig Ileal Smooth Muscle

 Materials
  • 200- to 450-g male Hartley guinea pig (Charles River Laboratories)
  • CO2 (i.e., gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 37° ± 1°C
  • Standard 5-HT receptor antagonists (e.g., granisetron; see recipe)
  • 1 mM methacholine bromide (see recipe)
  • 10 mM pargyline (see recipe)
  • Test compound(s)
  • Standard 5-HT receptor agonist (e.g., 5-HT; see recipe)
  • Euthanasia chamber (Harvard Apparatus)
  • Surgical instruments: scalpel, scissors
  • 10-ml syringes
  • Petri dishes
  • 13.8-cm Pasteur pipets (0.65-cm o.d.)
  • Cotton wool
  • 1.0 surgical silk threads
  • Fixed tissue holder (WPI)
  • Isotonic transducer (Hugo Sachs Elektronik, type 373)
  • 10-ml tissue bath (WPI) (also see unit 4.4) including:
    • Thermostatically regulated water circulator (WPI)
    • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)
Basic Protocol 8:  5-HT4 Receptor–Mediated Relaxation of Rat Esophageal Tunica Muscularis Mucosa
 Materials
  • 250- to 350-g male Sprague-Dawley rat (Charles River Laboratories)
  • CO2 (gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 32° ± 1°C
  • Standard 5-HT receptor antagonists (e.g., GR 113808; see recipe)
  • 10 mM pargyline (see recipe)
  • 1 mM carbachol (see recipe)
  • Standard 5-HT agonist (e.g., 5-HT; see recipe)
  • Test compound(s)
  • Euthanasia chamber (Harvard Apparatus)
  • Surgical instruments: scalpel, fine forceps, scissors
  • Petri dishes
  • 1.0 surgical silk threads
  • Fixed tissue holder with stainless steel wire mount (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue suspension wire
  • 10-ml tissue bath (WPI) (also see unit 4.4) including:
    • Thermostatically regulated water circulator (WPI)
    • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)
Alternate Protocol 2:  5-HT4 Receptor–Mediated Contraction of Guinea Pig Distal Colon
 Additional Materials (also see Basic Protocol 8)
  • 200- to 450-g male Dunkin Hartley guinea-pig (Charles River Laboratories)
  • 10-ml pipets
  • Squirt bottle
  • Glass rod (5-mm diameter)
  • Q-tips

Basic Protocol 9: Post-Junctional 5-HT7 Receptor–Mediated Relaxation of Guinea Pig Isolated Ileum

 Materials
  • 200- to 450-g male Dunkin-Hartley guinea pigs (Charles River Laboratories)
  • CO2 (i.e., gas cylinder or dry ice pellets to which a small amount of water has been added)
  • Krebs-Henseleit solution with and without corticosterone, cocaine, indomethacin, and atropine (see recipe), 37°C
  • 10 mM pargyline (see recipe)
  • Histamine or substance P
  • Standard 5-HT agonists (e.g., 5-CT, 5-HT; see recipe)
  • Test compound(s)
  • Standard 5-HT receptor antagonists (see recipe)
  • Euthanasia chamber (Harvard Apparatus)
  • Surgical instruments: scalpel, scissors
  • 10-ml syringes
  • 1.0 surgical silk threads
  • Petri dishes
  • Fixed tissue holder with stainless steel wire mount (WPI)
  • Isometric transducer (e.g., Grass FT03, Danish Myo Technology (DMT) Force transducer 750 TOBS) with stainless steel tissue suspension wire
  • 10-ml tissue bath (WPI) (also see unit 4.4) including:
    • Thermostatically regulated water circulator (WPI)
    • Polygraph/chart recorder/electronic data acquisition system (e.g., Biopac Acknowledge, Notocord, Danish Myo Technology)

Alternate Protocol 3: 5-HT7 Receptor–Mediated Relaxation of the Endothelium-Denuded, Precontracted Rabbit Jugular Vein

 Additional Materials (also see Basic Protocol 9)
  • 2.4- to 3.0-kg male New Zealand white rabbit (Froxfield, Hampshire, UK)
  • Pentobarbitone/sodium heparin (Henry Schein)
  • Krebs-Henseleit solution with and without corticosterone and cocaine (see recipe), 37° +/– 1°C
  • 1 mM phenoxybenzamine (see recipe)
  • Standard 5-HT receptor agonist (i.e., 0.1 mM -methyl-5-HT; see recipe)
  • Standard 5-HT receptor antagonists (e.g., SB-269970; see recipe)
  • Test compound(s)
  • 1 mM U-46619 (see recipe)
  • Standard 5-HT receptor agonists (e.g., 5-CT, 5-HT; see recipe)
  • 0.04 × ~3–in. long stainless steel wire roughened by rubbing with steel wool
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Figures

  • Figure 4.19.1
    Isolated, transmurally electrically stimulated guinea-pig ileum assay. (A) Details of the isolated tissue holder unit showing the position of the two platinum electrodes. (B) Diagram showing the tissue holder in position in the organ bath.

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  • Figure 4.19.2
    Treatment groups A, B, and C should be allocated according to a block design as illustrated, such that each day a different tissue bath is used for each treatment group.

    View Image
  • Figure 4.19.3
    5-Carboxyamidotryptamine (5-CT) concentration-response curves obtained in the absence (filled circles) and presence of 0.1 (open circles), 0.3 (filled squares), and 1 µM (open squares) (±)-WAY100135 on the isolated, transmurally electrically stimulated guinea pig ileum assay. Data redrawn with permission from Fletcher et al. (1993).

    View Image
  • Figure 4.19.4
    Dissection of rat caudal artery. (A) Details of how to appropriately cut the tail to expose the caudal artery. (B) Diagram showing the caudal artery and vein with indication as to the section to dissect.

    View Image
  • Figure 4.19.5
    Isolated rat caudal artery assay. (A) Details of the isolated tissue holder unit showing position of the artery ring on the tissue holder. (B) Diagram showing the tissue holder in position in the organ bath.

    View Image
  • Figure 4.19.6
    Contractile responses to 5-hydroxytryptamine (5-HT) in a single ring of rat caudal artery following pretreatment with 3 µM phenoxybenzamine and precontraction with 10 nM U46619. Data redrawn with permission from Craig and Martin (1993).

    View Image
  • Figure 4.19.7
    Diagram showing the area of cerebrum from which to remove the appropriate section of rabbit cerebral artery. Dissect the rostral cerebral artery from the caudal surface of the cerebellum with a pair of fine dissection scissors or scalpel. See arrows for region to dissect.

    View Image
  • Figure 4.19.8
    5-HT concentration-response curves obtained in the absence (filled squares) and presence of increasing (i.e., 1, 10, and 100 nM; open squares, filled circles, and open circles, respectively) concentrations of methiothepin on the isolated, rabbit cerebral arteries. Data redrawn with permission from Deckert et al. (1994).

    View Image
  • Figure 4.19.9
    5-HT concentration-response curves obtained in the absence (filled circles) and presence of 1 (open circles), 3 (filled squares), 10 (open squares), 30 (filled triangles), and 100 nM (open triangles) spiperone on the isolated rabbit thoracic aorta. Data redrawn with permission from Leff and Morse (1987).

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  • Figure 4.19.10
    5-HT concentration-response curves obtained in the absence (filled squares) and presence of 30 nM (open squares), 100 nM (filled circles), 300 nM (open circles), and 1 µM (filled diamonds) yohimbine, in the rat stomach fundus longitudinal muscle. Data redrawn with permission from Baxter et al. (1994).

    View Image
  • Figure 4.19.11
    5-HT concentration-response curves obtained in the absence (filled squares) and presence of 3 (open squares), 10 (filled circles), 30 (open circles), 100 (filled diamonds), and 300 µM (open diamonds) ,,-dimethyltryptamine in the rat jugular vein. The responses in this example data set are expressed as a percentage of the control curve maximum. Data redrawn with permission from Leff et al. (1986).

    View Image
  • Figure 4.19.12
    5-HT concentration-response curves obtained in the absence (filled squares) and presence of 30 (open squares), 100 (filled circles), and 300 nM (open circles) RS42358-197 in the guinea-pig ileum preparation. These data were obtained by a noncumulative dosing regimen on intact ileum segments and are expressed as a percentage of the control curve maximum. The Krebs solution contained 1 µM methysergide to block 5-HT1- and 5-HT2-receptors, and 10 µM 5-methoxytryptamine to desensitize 5-HT4 receptors. Data redrawn with permission from Eglen et al. (1993).

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  • Figure 4.19.13
    Preparation of the rat esophageal tunica muscularis mucosa. Schematic diagram showing the dissection procedure used to remove the external muscularis propria to expose the tunica muscularis.

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  • Figure 4.19.14
    5-HT concentration-response curves obtained in the absence (filled squares) and presence of 1 (open squares), 10 (filled circles), and 100 nM (open circles) GR113808 on the isolated rat esophagus. The responses in this example data set are expressed as a percentage of the control curve maximum. Data redrawn with permission from Gale et al. (1994).

    View Image
  • Figure 4.19.15
    5-HT concentration-response curves obtained in the absence (filled squares) and presence of increasing concentrations of 100 nM (open squares), 300 nM (filled circles), and 1 µM (open circles) SDZ 205 557 on the isolated guinea-pig distal colon. Effect is defined as the decrease in the force of contraction and is expressed as the fraction of 5-HT control maximum response. Data redrawn with permission from Wardle and Sanger (1993).

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  • Figure 4.19.16
    Representative polygraph trace showing the functional antagonism (relaxation) of 5-CT on contractures elicited by substance P in guinea pig ileum (used with permission from Carter et al., 1995).

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  • Figure 4.19.17
    5-HT concentration-response curves obtained in the absence (filled squares) and presence of increasing concentrations of 10 µM (open squares), 30 µM (filled circles), and 100 µM (open circles) N,N-dimethyltryptamine on endothelium-denuded rabbit jugular vein. Effect is defined as the decrease in the force of contraction and is expressed as the fraction of 5-HT control maximum response. Data redrawn with permission from Martin et al. (1987).

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    Leung, E., Walsh, L.K., Pulido-Rios, M.T., and Eglen, R.M. 1996b. Characterization of putative 5-HT7 receptors mediating direct relaxation in Cynomolgus monkey isolated jugular vein. Br. J. Pharmacol. 117:926-930.
    Martin, G.R., Leff, P., Cambridge, D., and Barrett, V.J. 1987. Comparative analysis of two types of 5-hydroxytryptamine receptor mediating vasorelaxation: Differential classification using tryptamines. N.-S. Arch. Pharmacol. 336:365-373.
    McLean, P.G. and Coupar I.M. 1995. 5-HT4 receptor antagonist affinities of SB207710, SB205008, and SB203186 in the human colon, rat oesophagus, and guinea-pig ileum peristaltic reflex. Naunyn Schmiedebergs Arch. Pharmacol. 352:132-140.
    Mize, A.L., Poisner, A.M., and Alper, R.H. 2001. Estrogens act in rat hippocampus and frontal cortex to produce rapid, receptor-mediated decreases in serotonin 5-HT1A receptor function. Neuroendocrinology 73:166-174.
    Oksenberg, D., Marsters, S.A., O'Dowd, B.F., Jin, H., Havlik, S., Peroutka, S.J., and Ashkenazi, A. 1992. A single amino-acid difference confers major pharmacological variation between human and rodent 5-HT1B receptors. Nature 360:161-163.
    Peroutka, S.J. and Snyder, S.H. 1979. Multiple serotonin receptors: Differential binding of [3H]5-hydroxytryptamine, [3H]lysergic acid diethylamide and [3H]spiroperidol. Mol. Pharmacol. 16:687-699.
    Prins, N.H., Briejer, M.R., Van Bergen, P.J., Akkermans, L.M., and Schuurkes, J.A. 1999. Evidence for 5-HT7 receptors mediating relaxation of human colonic circular smooth muscle. Br. J. Pharmacol. 128:849-852.
    Rapport, M.M., Green, A.A., and Page, I.H. 1948. Serum vasconstrictor (serotonin) IV isolation and characterization. J. Biol. Chem. 176:1243-1251.
    Ruat, M., Traiffort, E., Leurs, R., Tardivel-Lacombe, J., Diaz, J., Arrang, J., and Schwartz, J. 1993. Molecular cloning, characterization and localization of a high-affinity serotonin receptor (5-HT7) activating camp formation. Proc. Natl. Acad. Sci. U.S.A. 90:8547-8551.
    Shen, Y., Monsma, F.J., Metcalf, M.A., Jose, P.A., Hamblin, M.W., and Sibley, D.R. 1993. Molecular cloning and expression of a 5-hydroxytryptamine 7 serotonin receptor subtype. J. Biol. Chem. 268:18200-18204.
    Stollack, J.S. and Furchgott, R.F. 1983. Use of selective antagonists for determining the types of receptors mediating the actions of 5-hydroxytryptamine and tryptamine in the isolated rabbit aorta. J. Pharmacol. Exp. Therap. 224:215-221.
    Terrón, J.A. and Falcón-Neri, A. 1999. Pharmacological evidence for the 5-HT7 receptor mediating smooth muscle relaxation in canine cerebral arteries. Br. J. Pharmacol. 127:609-616.
    Tonini, M., Vicini, R., Cervio, E., De Ponti, F., De Giorgio, R., Barbara, G., Stanghellini, V., Dellabianca, A., and Sternini, C. 2005. 5-HT7 receptors modulate peristalsis and accomodation in the guinea pig ileum. Gastroenterology 129:1557-1566.
    Wardle, K.A. and Sanger, G.J. 1993. The guinea-pig distal colon—A sensitive preparation for the investigation of 5-HT4 receptor-mediated contractions. Br. J. Pharmacol. 13:1593-1599.
 Key References
    Alexander et al., 2008. See above.
    Bradley, P.B., Engel, G., Feniuk, W., Fozard, J.R., Humphrey, P.P.A., Middlemiss, D.N., Mylecharane, E.J., Richardson, B.P., and Saxena, P.R. 1986. Proposals for the classification and nomenclature of functional receptors for 5-hydroxytryptamine. Neuropharmacology 25:563-576.
    Gaddum and Picarelli, 1957. See above.
    Hoyer, D., Hannon, J.P., and Martin, G.R. 2002. Molecular, pharmacological and functional diversity of 5-HT receptors. Pharmacol. Biochem. Behav. 71:533-554.
    Humphrey et al., 1993. See above.
    Peroutka and Snyder, 1979. See above.
    Saxena, P.R., De Vries, P., and Villalon, C.M. 1998. 5-HT-like receptors: A time to bid goodbye. Trends Pharmacol. Sci. 19:311-316.

The above references are recommended for tracing the development of the current 5-HT receptor classification scheme and its associated nomenclature.

 Internet Resources
    http://www.iuphar-db.org/GPCR/ReceptorListForward

The above Web site is recommended for obtaining a comprehensive list of 5-HT receptor ligands and references.

     
 
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