Skip to main content
Download PDF

Effect of the 5-HT4 receptor agonist tegaserod on the expression of GRK2 and GRK6 in the rat gastrointestinal tract

BMC Research Notes volume 11, Article number: 362 (2018) Cite this article

Abstract

Objective

Tegaserod is a 5-hydroxytryptamine type 4 (5-HT4) receptor agonist, formerly used in treating constipation predominant irritable bowel syndrome, which desensitizes 5-HT4 receptors in rat oesophagus and colon in vitro. Desensitization of 5-HT4 receptors is regulated by G-protein coupled receptor kinases. This study was designed to assess the effect of 5-HT4 receptor activation on the expression of GRK2 and GRK6 in the rat oesophagus and distal colon by acute administration of tegaserod.

Results

Rats were treated with a single dose of tegaserod (5 mg/kg) and tissue samples of the oesophagus and distal colon were prepared and level of GRK2 and GRK6 protein expression was determined using western blotting. The immunodensity of GRK2 and GRK6 was normalized against the loading control β-actin and compared with control animals. Acute administration of tegaserod for 1, 2, 3, 4, 6, and 8 h did not change significantly the immunodensity of GRK2 or GRK6 in the oesophagus or GRK2 in the distal colon when compared with control animals. This may indicate that the basal level of GRK2 and GRK6 expression is sufficient to regulate the desensitization of 5-HT4 receptors in acute drug treatment.

Introduction

5-Hydroxytryptamine type 4 (5-HT4) receptor agonists have prokinetic effects in the gastrointestinal tract stimulating motility and secretion through enhanced acetylcholine release from excitatory motor neurons and interneurons [1]. Tegaserod is a 5-HT4 receptor agonist with clinical efficacy in patients with constipation-predominant irritable bowel syndrome and chronic constipation [2]. Tegaserod was used for the treatment of constipation-predominant irritable bowel syndrome in females and chronic constipation for both males and females until withdrawn in 2007 as it was associated with rare adverse cardiovascular effects [2, 3]. Tegaserod increases gastric emptying and accelerates small intestine and colonic transit in healthy human subjects [4] but increases colonic transit without altering gastric emptying in patients with constipation-predominant irritable bowel syndrome [5]. Tegaserod has a low therapeutic gain of 5–12% above placebo and is poorly absorbed with about two-thirds of oral doses being eliminated in the faeces [6]. It facilitates the peristaltic reflex in mouse, rat, guinea-pig and human intestine and also attenuates sensory neurotransmission in human rectum [7,8,9,10,11,12]. Tegaserod increased the amplitude of evoked excitatory postsynaptic currents in cultured myenteric neurons from mice, which desensitized rapidly, making it difficult to obtain responses to higher concentrations [13]. In addition, tegaserod desensitizes 5-HT4 receptors in rat oesophagus and colon in vitro [14, 15].

G protein coupled receptor (GPCR) kinase (GRK) regulated homologous desensitization has been reported for 5-HT4 receptors in mouse colliculus neurons and rat oesophagus and colon [14,15,16,17]. Cell culture studies using co-expression of GRKs with different 5-HT4 receptor splice variants showed the involvement of GRK2 and GRK5 in desensitization [18,19,20]. In contrast, desensitization of 5-HT4 receptors has been associated with GRK6 in the oesophagus and both GRK2 and GRK6 in the distal colon of the rat [15].

Changes in the level of GRK expression elicited by in vivo stimulation or blockade of GPCRs by agonist and antagonist is complex and not predictable [21, 22]. For example, chronic treatment with desipramine (a noradrenaline reuptake blocker) and acute treatment with fluoxetine (a 5-HT reuptake blocker) did not change significantly the immunodensity of GRK2/3 [23]. While chronic treatment with both opioid agonists and antagonists increases the expression of GRK2 and GRK6 in the brain [24, 25]. The molecular mechanisms associated with the regulation of 5-HT4 receptors in vivo by specific GRKs after treatment with 5-HT4 receptor agonist has not been studied. Therefore, the aim of this study was to assess the effect of 5-HT4 receptor activation on the expression level of GRK2 and GRK6 in the rat oesophagus and distal colon by acute administration of tegaserod.

Main text

Methods

Adult male Sprague–Dawley rats (Monash Animal Services) weighing 200–280 g were randomly divided into six treatment groups and one control group (n = 6; total number = 42). Rats were housed at the Faculty of Pharmacy and Pharmaceutical Sciences, Monash University’s animal facility in a 12 h light–dark cycle with food and water ad libitum. Rats were treated with a single intraperitoneal injection of tegaserod maleate (Sequoia Research Products Ltd, Berkshire, UK) (5 mg/kg) dissolved in vehicle (10% sulfobutyl ether-beta cyclodextrin) or just vehicle (controls). Rats were killed 1 h after vehicle or 1, 2, 3, 4, 6 and 8 h after drug administration by carbon dioxide asphyxiation. Animals were quickly dissected and the lower third of the oesophagus proximal to the diaphragm, and the distal colon were excised. The outer muscularis externa of the oesophagus was separated from the inner tunica muscularis mucosae. Tissues were washed three times with ice-cold phosphate buffered saline (PBS) and homogenized in 1:10 (w/v) of homogenization buffer [50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 1% Nonidet P40, 0.5% sodium deoxycholate, complete protease inhibitor cocktail tablet (Roche, Sydney, Australia)] for 1 min before centrifugation at 12,000×g for 10 min. Supernatant protein concentration was determined by Quant-It protein assay (Invitrogen) and homogenates were stored at − 80 °C until used.

Total protein (50 µg) was separated on 12% polyacrylamide gels by SDS-PAGE and transferred to nitrocellulose membranes (Amersham Bioscience). Each gel was run with a sample from control to all test groups. Nonspecific binding to the membrane was blocked with Odyssey blocking buffer (LI-COR Biosciences) for 1 h. Membranes were probed with combination of two primary antibodies: goat anti-β-actin and either mouse anti-GRK2 or rabbit anti-GRK6 (Table 1) overnight at 4 °C and washed four times with PBS and 0.1% Tween 20 PBS (PBST). Membranes were incubated with fluorophore-conjugated secondary antibodies (Table 1) for 1 h at room temperature and washed four times for five min using PBST. An Odyssey Infrared Imaging System (LI-COR Biosciences) was used to examine the immunoblots. The integrated optical density of the immunoreactivity was assessed using the Odyssey Infrared Imaging System software (LI-COR Biosciences). The integral optical density of GRK2 and GRK6 from each lane was normalised against β-actin for that lane and expressed as percentage of control values.

Table 1 Details of primary and secondary antibodies
Full size table

Data for each treatment group was expressed as mean ± standard error of mean (SEM) of GRK2 and GRK6 levels expressed as percentage of control values. Statistical analyses were performed using GraphPad Prism 5 (GraphPad Software, La Jolla, California USA). Values were compared between the different groups using one-way analysis of variance (ANOVA) followed with Dunnett’s multiple comparison post hoc test. A P value < 0.05 was considered to be statistically significant.

Results

Immunoblot analysis of the tissue homogenate of the oesophagus and distal colon revealed immunoreactive protein of around 80 kDa for GRK2 or alternatively 66 kDa for GRK6 under the 800 nm excitation wavelength channel and around 42 kDa for β-actin under the 700 nm excitation wavelength channel on the same membrane (Fig. 1). This allowed the analysis of the immunodensity of GRK2 and β-actin or GRK6 and β-actin on the same membrane. The immunodensity of GRK2 relative to β-actin did not change significantly in either the oesophagus (P = 0.52, one-way ANOVA) or distal colon (P = 0.66, one-way ANOVA) following acute treatment with tegaserod (Fig. 1a, b). Similarly, the immunodensity of GRK6 relative to β-actin in the oesophagus (Fig. 1c) not change significantly (P = 0.92, one-way ANOVA) following acute treatment with tegaserod.

Fig. 1
figure 1

Time course study of the effect of tegaserod (5 mg/kg, i.p.) on the expression levels of GRK2 and GRK6 in the rat digestive tract. a GRK2 levels in the oesophagus; b GRK2 levels in the distal colon; and c GRK6 levels in the oesophagus. Upper panels: representative immunoblots illustrating the effects of tegaserod on the expression level of GRK2 or GRK6 at 1, 2, 3, 4, 6 and 8 h after treatment. Lower panels: the mean densitometric values of GRK2 or GRK6 levels relative to β-actin and expressed as percentages of values in untreated control rats (± SEM, n = 6 per group). No significant difference from the control occurred in any of the treatments (P > 0.05; one-way ANOVA). Con control, h hours following treatment

Full size image

Discussion

This study was designed to investigate the effect of acute activation of 5-HT4 receptors using the 5-HT4 receptor agonist tegaserod on the expression level of GRK2 and GRK6 in rat oesophagus and distal colon. Prior studies indicate that in the oesophagus GRK6 is associated with the desensitization of 5-HT4 receptors whilst GRK2 and GRK6 are associated with desensitization of 5-HT4 receptors in the distal colon of the rat [15]. GRK-mediated GPCR desensitization has a physiological significance to protect cells from over-stimulation in the persistent presence of agonists and to keep the signals under regulation. Up-regulation of GRKs facilitates GPCR desensitization whilst down-regulation of GRKs impedes GPCR desensitization in vitro and in vivo [26]. Intravenous administration of tegaserod (0.001–1 mg/kg) has been reported to evoke a dose-dependent increase in relaxation of the longitudinal muscle of the rat oesophagus. The response had a rapid onset (commencing less than 1 min after dosing) and reached a maximum typically within 2 min [27,28,29]. In addition, subcutaneous administration of tegaserod (0.03, 0.3 and 3 mg/kg) has been reported to produce a dose-dependent colonic prokinetic effect in guinea-pigs [30]. Moreover, intraperitoneal administration of tegaserod (0.1, 1, 10 mg/kg) has been reported to produce a dose-dependent reduction in the number of abdominal contractions induced by colonic distensions in rats [8]. Based on the above studies a single dose of tegaserod (5 mg/kg i.p.) was used for time course study of the effect of 5-HT4 receptor activation on the level of expression of GRK2 and GRK6 in the rat oesophagus and distal colon.

Similar to the finding of the present in vivo study, acute treatment with the 5-HT reuptake blocker fluoxetine that increases the synaptic concentration of 5-HT, was not associated with changes in the immunodensity of GRK2/3 in rat frontal cortex membrane and cytosolic fractions [23]. In contrast, acute treatment with tricyclics antidepressant desipramine increased in a dose and time-dependent manner the content of GRK2/3 in the membrane whilst chronic treatment did not alter the immunodensity of GRK2/3 [23].

Tegaserod activates 5-HT4 receptors on intrinsic primary afferent neurons and mimics the action of endogenous serotonin released from enterochromaffin cells. It stimulates the intrinsic primary afferent neurones that release transmitters such as calcitonin gene-related peptide, which activate cholinergic interneurons. The cholinergic interneurons activate the peristaltic reflex [9]. Tegaserod has been shown to desensitize the peristaltic reflex induced by mucosal stimulation in a time and concentration dependent manner in rat colon [14]. In addition, in rat oesophagus, tegaserod desensitized 5-HT induced-relaxation of the tunica muscularis mucosae in a time-dependent manner [15]. The lack of change in the expression level of GRK2 in the oesophagus (Fig. 1a) is in line with the immunohistochemistry study showing that there was no expression of GRK2 on the smooth muscle of the muscularis mucosae where the 5-HT4 receptors are concentrated in the oesophagus [15, 31, 32]. The interesting finding is the lack of change in the expression level of GRK6 in the oesophagus and GRK2 in distal colon tissues (Fig. 1b, c) where they were found to be co-expressed and co-immunoprecipitated with 5-HT4 receptors [15]. This may be due to the expression of sufficient levels of GRK2 and GRK6 at the basal level to regulate the desensitization of 5-HT4 receptor. Alternatively, it may require chronic administration of tegaserod to obtain changes in the expression level of GRK2 and GRK6. The desensitization of GPCRs requires translocation of GRK2 from cytosol to membrane upon activation by agonist [33]. Analysis of change in the immunoreactivity of GRK2 in the membrane fraction warrant further investigation.

In the distal colon, 5-HT4 receptor immunoreactive cells were found in longitudinal muscle, myenteric plexuses, circular muscle, submucosal plexuses and muscularis mucosae. GRK6 was expressed in the longitudinal muscle, circular muscle, and muscularis mucosae and co-immunoprecipitated with 5-HT4 receptors [15]. Due to the interference of the background it was not possible to quantify the immunodensity of GRK6. In addition, the 5-HT4 receptors are located presynaptically at neuronal synapses within the myenteric plexus [1] and GRK5 is exclusively localised on the nerve endings of both myenteric and submucosal plexuses [15]. It is therefore worthwhile to investigate the effect of tegaserod on the expression of GRK5 and GRK6 at mRNA level in the distal colon.

Based on desensitization studies of β2-adrenoceptors and M3 muscarinic receptors, it is often generalized that partial agonists induce less desensitization of GPCRs than full agonists. They could stabilize receptor conformations that differ in their capacity to interact or serve as substrates for GRKs and arrestins to generate downstream recognition barcodes [34, 35]. Partial agonists augment submaximal endogenous stimulation and prevent an exaggerated response to an endogenous agonist. As a result partial agonists have a lower tendency to induce receptor desensitization and/or receptor down-regulation [34, 35]. However, the capacity of agonists to induce desensitization of 5-HT4 receptors depends more on the activation potency of the drug than its efficacy [16]. The lower potency of tegaserod may contribute to the lack of its effect on the expression level of GRKs in rat oesophagus and distal colon. Alternatively, tegaserod has similar binding affinities to both 5-HT4 and 5-HT2B receptors [30] and acts as an antagonist at the 5-HT2B receptor [29, 30]. Thus, the 5-HT2B receptor antagonist effect of tegaserod may counteract its effect on the expression level of GRKs.

Limitations

Limitations of this study include the use of only one concentration of tegaserod. However, the concentration used reduces the number of abdominal contractions induced by colonic distensions in rats measured over 90 min [8]. Taken together with prior observations that desensitization is only detectable after 20 min in rat oesophagus [15], our sampling times of 1–8 h should detect changes in GRK expression. Another limitation is the use of a control at only a single time point, which occurred in part to restrict animal usage. A further limitation is the lack of positive control for changes in GRK2 and 6 expression although our prior study [15] indicated that the antisera used detected changes in the expression level of these proteins. So far, no studies have investigated the effect of 5-HT4 receptor activation on the expression level of GRKs using agonists with different potency, selectivity and efficacy. Such studies will help understand whether the expression of GRKs is altered based on agonist potency, selectivity or efficacy in physiological systems.

Abbreviations

5-HT4 :

5-hydroxytryptamine type 4

ANOVA:

one-way analysis of variance

GPCR:

G-protein coupled receptor

GRK:

GPCR kinase

PBS:

phosphate buffered saline

PBST:

phosphate buffered saline Tween

SEM:

standard error of mean

References

  1. Gershon MD, Tack J. The serotonin signaling system: from basic understanding to drug development for functional GI disorders. Gastroenterology. 2007;132:397–414.

    Article  PubMed  CAS  Google Scholar 

  2. Beattie DT, Smith JAM. Serotonin pharmacology in the gastrointestinal tract: a review. Naunyn-Schmiedeberg’s Arch Pharmacol. 2008;377:181–203.

    Article  CAS  Google Scholar 

  3. McLean PG, Borman RA, Lee K. 5-HT in the enteric nervous system: gut function and neuropharmacology. Trends Neurosci. 2007;30:9–13.

    Article  PubMed  CAS  Google Scholar 

  4. Degen L, Matzinger D, Merz M, Appel-Dingemanse S, Osborne S, Luchinger S, Bertold R, Maecke H, Beglinger C. Tegaserod, a 5-HT4 receptor partial agonist, accelerates gastric emptying and gastrointestinal transit in healthy male subjects. Aliment Pharmacol Ther. 2001;15:1745–51.

    Article  PubMed  CAS  Google Scholar 

  5. Prather CM, Camilleri M, Zinsmeister AR, McKinzie S, Thomforde G. Tegaserod accelerates orocecal transit in patients with constipation-predominant irritable bowel syndrome. Gastroenterology. 2000;118:463–8.

    Article  PubMed  CAS  Google Scholar 

  6. De Maeyer JH, Lefebvre RA, Schuurkes JAJ. 5-HT4 receptor agonists: similar but not the same. Neurogastroenterol Motil. 2008;20:99–112.

    Article  PubMed  CAS  Google Scholar 

  7. Coffin B, Farmachidi JP, Rueegg P, Bastie A, Bouhassira D. Tegaserod, a 5-HT4 receptor partial agonist, decreases sensitivity to rectal distension in healthy subjects. Aliment Pharmacol Ther. 2003;17:577–85.

    Article  PubMed  CAS  Google Scholar 

  8. Greenwood-Van Meerveld B, Venkova K, Hicks G, Dennis E, Crowell MD. Activation of peripheral 5-HT4 receptors attenuates colonic sensitivity to intraluminal distension. Neurogastroenterol Motil. 2006;18:76–86.

    Article  PubMed  CAS  Google Scholar 

  9. Grider JR, Foxx-Orenstein AE, Jin JG. 5-Hydroxytryptamine4 receptor agonists initiate the peristaltic reflex in human, rat, and guinea pig intestine. Gastroenterology. 1998;115:370–80.

    Article  PubMed  CAS  Google Scholar 

  10. Hoffman JM, Tyler K, MacEachern SJ, Balemba OB, Johnson AC, Brooks EM, Zhao H, Swain GM, Moses PL, Galligan JJ, et al. Activation of colonic mucosal 5-HT4 receptors accelerates propulsive motility and inhibits visceral hypersensitivity. Gastroenterology. 2012;142(844–54):e4.

    Google Scholar 

  11. Ji SW, Park H, Chung JP, Lee SI, Lee YH. Effects of tegaserod on ileal peristalsis of guinea pig in vitro. J Pharmacol Sci. 2004;94:142–52.

    Article  Google Scholar 

  12. McGuire C, Boundouki G, Hockley JRF, Reed D, Cibert-Goton V, Peiris M, Kung V, Broad J, Aziz Q, Chan C, et al. Ex vivo study of human visceral nociceptors. Gut. 2017;67:86–96.

    Article  Google Scholar 

  13. Liu M, Geddis MS, Wen Y, Setlik W, Gershon MD. Expression and function of 5-HT4 receptors in the mouse enteric nervous system. Am J Physiol Gastrointest Liver Physiol. 2005;289:G1148–63.

    Article  PubMed  CAS  Google Scholar 

  14. Grider JR. Desensitization of the peristaltic reflex induced by mucosal stimulation with the selective 5-HT4 agonist tegaserod. Am J Physiol. 2006;290:319–27.

    Google Scholar 

  15. Nedi T, White PJ, Coupar IM, Irving HR. Tissue dependent differences in G-protein coupled receptor kinases associated with 5-HT4 receptor desensitization in the rat gastro-intestinal tract. Biochem Pharmacol. 2011;81:123–33.

    Article  PubMed  CAS  Google Scholar 

  16. Ansanay H, Sebben M, Bockaert J, Dumuis A. Characterization of homologous 5-hydroxytryptamine4 receptor desensitization in colliculi neurons. Mol Pharmacol. 1992;42:808–16.

    PubMed  CAS  Google Scholar 

  17. Rondé P, Ansanay H, Dumuis A, Miller R, Bockaert J. Homologous desensitization of 5-hydroxytryptamine4 receptors in rat esophagus: functional and second messenger studies. J Pharmacol Exp Ther. 1995;272:977–83.

    PubMed  Google Scholar 

  18. Barthet G, Carrat G, Cassier E, Barker B, Gaven F, Pillot M, Framery B, Pellissier LP, Augier J, Kang DS, et al. Β-arrestin1 phosphorylation by GRK5 regulates G protein-independent 5-HT4 receptor signalling. EMBO J. 2009;28:2706–18.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Barthet G, Gaven F, Framery B, Shinjo K, Nakamura T, Claeysen S, Bockaert J, Dumuis A. Uncoupling and endocytosis of 5-hydroxytryptamine 4 receptors: distinct molecular events with different GRK2 requirements. J Biol Chem. 2005;280:27924–34.

    Article  PubMed  CAS  Google Scholar 

  20. Mnie-Filali O, Amraei MG, Benmbarek S, Archer-Lahlou E, Peñas-Cazorla R, Vilaró MT, Boye SM, Piñeyro G. Serotonin 4 receptor (5-HT4R) internalization is isoform-specific: effects of 5-HT and RS67333 on isoforms A and B. Cell Signal. 2010;22:501–9.

    Article  PubMed  CAS  Google Scholar 

  21. Ahmed MR, Gurevich V, Dalby KN, Benovic JL, Gurevich EV. Haloperidol and clozapine differentially affect the expression of arrestins, receptor kinases, and extracellular signal-regulated kinase activation. J Pharmacol Exp Ther. 2008;325:276–83.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  22. Fan X, Zhang J, Zhang X, Yue W, Ma L. Acute and chronic morphine treatments and morphine withdrawal differentially regulate GRK2 and GRK5 gene expression in rat brain. Neuropharmacol. 2002;43:809–16.

    Article  CAS  Google Scholar 

  23. Miralles A, Asensio VJ, Garcia-Sevilla JA. Acute treatment with the cyclic antidepressant desipramine, but not fluoxetine, increases membrane-associated G protein-coupled receptor kinases 2/3 in rat brain. Neuropharmacol. 2002;443:1249–57.

    Article  Google Scholar 

  24. Diaz A, Pazos A, Florez J, Ayesta FJ, Santana V, Hurle MA. Regulation of mu-opioid receptors, G-protein-coupled receptor kinases and beta-arrestin 2 in the rat brain after chronic opioid receptor antagonism. Neuroosci. 2002;112:345–53.

    Article  CAS  Google Scholar 

  25. Hurle MA. Changes in the expression of G protein-coupled receptor kinases and beta-arrestin 2 in rat brain during opioid tolerance and supersensitivity. J Neurochem. 2001;77:486–92.

    Article  PubMed  CAS  Google Scholar 

  26. Gainetdinov RR, Premont RT, Bohn LM, Lefkowitz RJ, Caron MG. Desensitization of G protein-coupled receptors and neuronal functions. Annu Rev Neurosci. 2004;27:107–44.

    Article  PubMed  CAS  Google Scholar 

  27. Armstrong SR, McCullough JL, Beattie DT. Measurement of 5-HT4 receptor-mediated esophageal responses by digital sonomicrometry in the anesthetized rat. J Pharmacol Toxicol Meth. 2006;53:198–205.

    Article  CAS  Google Scholar 

  28. Beattie DT, Armstrong SR, Shaw JP, Marquess D, Sandlund C, Smith JA, Taylor JA, Humphrey PP. The in vivo gastrointestinal activity of TD-5108, a selective 5-HT(4) receptor agonist with high intrinsic activity. Naunyn-Schmiedeberg’s Arch Pharmacol. 2008;378:139–47.

    Article  CAS  Google Scholar 

  29. McCullough JL, Armstrong SR, Hegde SS, Beattie DT. The 5-HT2B antagonist and 5-HT4 agonist activities of tegaserod in the anaesthetized rat. Pharmacol Res. 2006;53:353–8.

    Article  PubMed  CAS  Google Scholar 

  30. Beattie DT, Smith JA, Marquess D, Vickery RG, Armstrong SR, Pulido-Rios T, McCullough JL, Sandlund C, Richardson C, Mai N, Humphrey PP. The 5-HT4 receptor agonist, tegaserod, is a potent 5-HT2B receptor antagonist in vitro and in vivo. Br J Pharmacol. 2004;143:549–60.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Baxter GS, Craig DA, Clarke DE. 5-Hydroxytryptamine4 receptors mediate relaxation of the rat oesophageal tunica muscularis mucosae. Naunyn-Schmiedeberg’s Arch Pharmacol. 1991;343:439–46.

    Article  CAS  Google Scholar 

  32. Poole DP, Xu B, Koh SL, Hunne B, Coupar IM, Irving HR, Shinjo K, Furness JB. Identification of neurons that express 5-hydroxytryptamine-4 receptors in the intestine. Cell Tissue Res. 2006;325:413–22.

    Article  PubMed  CAS  Google Scholar 

  33. Premont RT, Gainetdinov RR. Physiological roles of G protein-coupled receptor kinases and arrestins. Annu Rev Physiol. 2007;69:511–34.

    Article  PubMed  CAS  Google Scholar 

  34. Bahouth SW, Nooh MM. Barcoding of GPCR trafficking and signaling through the various trafficking roadmaps by compartmentalized signaling networks. Cell Signal. 2017;36:42–55.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  35. Clark RB, Knoll BJ, Barber R. Partial agonists and G protein-coupled receptor desensitization. Trends Pharmacol Sci. 1999;20:279–86.

    Article  PubMed  CAS  Google Scholar 

  36. Burgos-Ramos E, Puebla-Jimenez L, Arilla-Ferreiro E. Minocycline provides protection against beta-amyloid(25-35)-induced alterations of the somatostatin signaling pathway in the rat temporal cortex. Neurooscience. 2008;154:1458–66.

    Article  CAS  Google Scholar 

  37. Matuzany-Ruban A, Golan M, Miroshnik N, Schreiber G, Avissar S. Normalization of GRK2 protein and mRNA measures in patients with depression predict response to antidepressants. Int J Neuropsychopharmacol. 2009;2009:1–9.

    Google Scholar 

  38. Wu H, Wei W, Song L, Zhang L, Chen Y, Hu X. Paeoniflorin induced immune tolerance of mesenteric lymph node lymphocytes via enhancing beta 2-adrenergic receptor desensitization in rats with adjuvant arthritis. Int Immunopharmacol. 2007;7:662–73.

    Article  PubMed  CAS  Google Scholar 

  39. Dzimiri N, Muiya P, Andres E, Al-Halees Z. Differential functional expression of human myocardial G protein receptor kinases in left ventricular cardiac diseases. Eur J Pharmacol. 2004;489:162–77.

    Article  CAS  Google Scholar 

  40. Grange-Midroit M, Garcia-Sevilla JA, Ferrer-Alcon M, La Harpe R, Walzer C, Guimon J. G protein-coupled receptor kinases, beta-arrestin-2 and associated regulatory proteins in the human brain: postmortem changes, effect of age and subcellular distribution. Brain Res Mol Brain Res. 2002;101:39–51.

    Article  PubMed  CAS  Google Scholar 

Download references

Authors’ contributions

TN designed and undertook the experiments. TN, IC, PW and HI were involved in data analyses and interpretation. TN and HI drafted the manuscript. All authors read and approved the final manuscript.

Acknowledgements

The authors are grateful to Dr. RJ Prankerd (Monash University) for providing the SBE/CD.

Competing interests

The authors declare that they have no competing interests.

Availability of data and materials

All data generated or analysed in this study are included in the published article.

Consent for publication

Not applicable.

Ethics approval and consent to participate

All procedures conformed to Australian National Health and Medical Research Council’s guidelines and were approved by the Victorian College of Pharmacy Animal Ethics Committee of Monash University. Rats were housed at the Faculty of Pharmacy and Pharmaceutical Sciences, Monash University’s animal facility in a 12 h light–dark cycle with food and water ad libitum.

Funding

The Faculty of Pharmacy and Pharmaceutical Sciences, Monash University supported the experimental work and provided a PhD stipend to TN.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Author information

Authors and Affiliations

  1. Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), 381 Royal Parade, Parkville, VIC, 3052, Australia

    Teshome Nedi, Paul J. White, Ian M. Coupar & Helen R. Irving

  2. School of Pharmacy, College of Health Sciences, Addis Ababa University, PO Box 1176, Addis Ababa, Ethiopia

    Teshome Nedi

  3. La Trobe Institute for Molecular Science, La Trobe University, PO Box 199, Bendigo, VIC, 3552, Australia

    Helen R. Irving

Authors
  1. Teshome Nedi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paul J. White
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ian M. Coupar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Helen R. Irving
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Helen R. Irving.

Rights and permissions

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Nedi, T., White, P.J., Coupar, I.M. et al. Effect of the 5-HT4 receptor agonist tegaserod on the expression of GRK2 and GRK6 in the rat gastrointestinal tract. BMC Res Notes 11, 362 (2018). https://doi.org/10.1186/s13104-018-3495-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s13104-018-3495-2

Keywords

  • Colon
  • Oesophagus
  • 5-HT4 receptors
  • G protein coupled receptor kinases
  • Receptor desensitization

BMC Research Notes

ISSN: 1756-0500

Contact us