Guanylin, Uroguanylin and Guanylate Cyclase-C Are Expressed in the Gastrointestinal Tract of Horses.
Abstract: Guanylate cyclase-C (GC-C) is a multifunctional receptor encoded by the gene, representing an attractive target for therapy in several gastrointestinal diseases in humans. Little is known about this system in horses. We investigated for the first time the gene expression of guanylin, uroguanylin and GC-C receptors in different horse's gastrointestinal tracts. Tissue samples from stomach, duodenum, jejunum, ileum, head and body of cecum, left and right dorsal colon, left and right ventral colon, pelvic flexure, transverse colon, descending colon and rectum were collected from adult horses within 1 h . For each sample, total RNA was extracted from 100 mg of ground tissue, and qRT-PCR performed on , and transcripts on a CFX96 Touch instrument. Data analysis was carried out with Bio-Rad CFX Manager software, and genes of interest normalized relative to the abundance of the two reference genes (). Additionally, the protein expression levels of GC-C receptor were analyzed through western blotting. A common pattern of expression throughout the gastrointestinal lumen for all three investigated transcripts was found. The expression of , and genes was higher in jejunum, ileum, descending colon and rectum. The levels of expression of GC-C protein confirmed these data. The findings of this study might open new scenarios for the therapeutic approach to enteric diseases of horse using selective agonists of GC-C.
Copyright © 2019 Cappelli, Gialletti, Tesei, Bassotti, Fettucciari, Capomaccio, Bonfili, Cuccioloni, Eleuteri, Spaterna and Laus.
Publication Date: 2019-09-27 PubMed ID: 31611814PubMed Central: PMC6776823DOI: 10.3389/fphys.2019.01237Google Scholar: Lookup
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Summary
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The research paper details the study of gene expression of guanylin, uroguanylin, and Guanylate cyclase-C (GC-C) receptors, three components involved in gastrointestinal diseases, in different areas of the horse’s gastrointestinal tract. This is the first time such a study has been conducted and it opens the potential for new therapies for equine enteric diseases.
Studying GC-C and Its Ligands in Horses
- Guanylate cyclase-C (GC-C) is a receptor that can perform multiple functions and is encoded by the GUCY2C gene. It is particularly interesting for scientists because it can be a target for therapy in multiple gastrointestinal diseases in human beings.
- In spite of its significance, there has been little research on this receptor in horses, which is what the authors of this paper set out to investigate. They specifically look at the gene expression of guanylin, uroguanylin, which are ligands, and GC-C receptors in the gastrointestinal tract of horses.
Methodology of the Study
- From adult horses the researchers collected tissue samples, within one hour of extraction, from various parts of the gastrointestinal tract, including the stomach, duodenum, and different parts of the colon and rectum.
- Using 100mg of ground tissue from each sample, they extracted total RNA. Subsequently, quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was used to analyze guanylin, uroguanylin, and GC-C receptor transcripts, utilizing a CFX96 Touch instrument.
- The Bio-Rad CFX Manager software was utilized to conduct data analysis. The genes of interest were examined relative to the abundance of two reference genes.
- Additionally, the protein expression levels of the GC-C receptor were examined through western blotting, another analytical technique.
Findings of the Study and Future Applications
- The researchers found a similar pattern of expression throughout the entire gastrointestinal lumen for all three transcripts they examined.
- The expression of guanylin, uroguanylin, and GUCY2C genes was shown to be higher in the jejunum, ileum, descending colon, and rectum. The protein expression levels of GC-C confirmed these findings.
- These insights could potentially open new avenues for therapy for enteric diseases in horses. This study’s findings suggest that these diseases could be treated with selective agonists of GC-C.
Cite This Article
APA
Cappelli K, Gialletti R, Tesei B, Bassotti G, Fettucciari K, Capomaccio S, Bonfili L, Cuccioloni M, Eleuteri AM, Spaterna A, Laus F.
(2019).
Guanylin, Uroguanylin and Guanylate Cyclase-C Are Expressed in the Gastrointestinal Tract of Horses.
Front Physiol, 10, 1237.
https://doi.org/10.3389/fphys.2019.01237 Publication
Researcher Affiliations
- Department of Veterinary Medicine, University of Perugia, Perugia, Italy.
- Department of Veterinary Medicine, University of Perugia, Perugia, Italy.
- School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.
- Department of Medicine, School of Medicine, University of Perugia, Perugia, Italy.
- Department of Experimental Medicine, School of Medicine, University of Perugia, Perugia, Italy.
- Department of Veterinary Medicine, University of Perugia, Perugia, Italy.
- School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.
- School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.
- School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.
- School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.
- School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.
References
This article includes 59 references
- Ahsan MK, Tchernychev B, Kessler MM, Solinga RM, Arthur D, Linde CI, Silos-Santiago I, Hannig G, Ameen NA. Linaclotide activates guanylate cyclase-C/cGMP/protein kinase-II-dependent trafficking of CFTR in the intestine.. Physiol Rep 2017 Jun;5(11).
- Aka AA, Rappaport JA, Pattison AM, Sato T, Snook AE, Waldman SA. Guanylate cyclase C as a target for prevention, detection, and therapy in colorectal cancer.. Expert Rev Clin Pharmacol 2017 May;10(5):549-557.
- Al-Salama ZT, Syed YY. Plecanatide: First Global Approval.. Drugs 2017 Apr;77(5):593-598.
- Andresen V, Camilleri M, Busciglio IA, Grudell A, Burton D, McKinzie S, Foxx-Orenstein A, Kurtz CB, Sharma V, Johnston JM, Currie MG, Zinsmeister AR. Effect of 5 days linaclotide on transit and bowel function in females with constipation-predominant irritable bowel syndrome.. Gastroenterology 2007 Sep;133(3):761-8.
- Andrusier N, Nussinov R, Wolfson HJ. FireDock: fast interaction refinement in molecular docking.. Proteins 2007 Oct 1;69(1):139-59.
- Bassotti G, Blandizzi C. Understanding and treating refractory constipation.. World J Gastrointest Pharmacol Ther 2014 May 6;5(2):77-85.
- Blikslager AT. Obstructive disorders of the gastrointestinal tract. Equine Internal Medicine 3rd Edn, 882–892.
- Blomain ES, Pattison AM, Waldman SA. GUCY2C ligand replacement to prevent colorectal cancer.. Cancer Biol Ther 2016 Jul 2;17(7):713-8.
- Brachelente C, Cappelli K, Capomaccio S, Porcellato I, Silvestri S, Bongiovanni L, De Maria R, Verini Supplizi A, Mechelli L, Sforna M. Transcriptome Analysis of Canine Cutaneous Melanoma and Melanocytoma Reveals a Modulation of Genes Regulating Extracellular Matrix Metabolism and Cell Cycle.. Sci Rep 2017 Jul 25;7(1):6386.
- Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 1976 May 7;72:248-54.
- Brenna Ø, Bruland T, Furnes MW, Granlund Av, Drozdov I, Emgård J, Brønstad G, Kidd M, Sandvik AK, Gustafsson BI. The guanylate cyclase-C signaling pathway is down-regulated in inflammatory bowel disease.. Scand J Gastroenterol 2015;50(10):1241-52.
- Brierley SM. Guanylate cyclase-C receptor activation: unexpected biology.. Curr Opin Pharmacol 2012 Dec;12(6):632-40.
- Busby RW, Bryant AP, Bartolini WP, Cordero EA, Hannig G, Kessler MM, Mahajan-Miklos S, Pierce CM, Solinga RM, Sun LJ, Tobin JV, Kurtz CB, Currie MG. Linaclotide, through activation of guanylate cyclase C, acts locally in the gastrointestinal tract to elicit enhanced intestinal secretion and transit.. Eur J Pharmacol 2010 Dec 15;649(1-3):328-35.
- Capomaccio S, Cappelli K, Barrey E, Felicetti M, Silvestrelli M, Verini-Supplizi A. Microarray analysis after strenuous exercise in peripheral blood mononuclear cells of endurance horses.. Anim Genet 2010 Dec;41 Suppl 2:166-75.
- Capomaccio S, Cappelli K, Spinsanti G, Mencarelli M, Muscettola M, Felicetti M, Verini Supplizi A, Bonifazi M. Athletic humans and horses: comparative analysis of interleukin-6 (IL-6) and IL-6 receptor (IL-6R) expression in peripheral blood mononuclear cells in trained and untrained subjects at rest.. BMC Physiol 2011 Jan 21;11:3.
- Capomaccio S, Vitulo N, Verini-Supplizi A, Barcaccia G, Albiero A, D'Angelo M, Campagna D, Valle G, Felicetti M, Silvestrelli M, Cappelli K. RNA sequencing of the exercise transcriptome in equine athletes.. PLoS One 2013;8(12):e83504.
- Cappelli K, Felicetti M, Capomaccio S, Nocelli C, Silvestrelli M, Verini-Supplizi A. Effect of training status on immune defence related gene expression in Thoroughbred: are genes ready for the sprint?. Vet J 2013 Mar;195(3):373-6.
- Cappelli K, Felicetti M, Capomaccio S, Pieramati C, Silvestrelli M, Verini-Supplizi A. Exercise-induced up-regulation of MMP-1 and IL-8 genes in endurance horses.. BMC Physiol 2009 Jun 24;9:12.
- Cappelli K, Felicetti M, Capomaccio S, Spinsanti G, Silvestrelli M, Supplizi AV. Exercise induced stress in horses: selection of the most stable reference genes for quantitative RT-PCR normalization.. BMC Mol Biol 2008 May 19;9:49.
- Danaee H, Kalebic T, Wyant T, Fassan M, Mescoli C, Gao F, Trepicchio WL, Rugge M. Consistent expression of guanylyl cyclase-C in primary and metastatic gastrointestinal cancers.. PLoS One 2017;12(12):e0189953.
- Forte LR. Guanylin regulatory peptides: structures, biological activities mediated by cyclic GMP and pathobiology.. Regul Pept 1999 May 31;81(1-3):25-39.
- Giannella RA, Mann EA. E. coli heat-stable enterotoxin and guanylyl cyclase C: new functions and unsuspected actions.. Trans Am Clin Climatol Assoc 2003;114:67-85; discussion 85-6.
- Hamra FK, Krause WJ, Eber SL, Freeman RH, Smith CE, Currie MG, Forte LR. Opossum colonic mucosa contains uroguanylin and guanylin peptides.. Am J Physiol 1996 Apr;270(4 Pt 1):G708-16.
- Hannig G, Tchernychev B, Kurtz CB, Bryant AP, Currie MG, Silos-Santiago I. Guanylate cyclase-C/cGMP: an emerging pathway in the regulation of visceral pain.. Front Mol Neurosci 2014;7:31.
- Hanson RR. Diseases of the large colon that can result in colic. Manual of Equine Gastroenterology 279–297.
- Javsicas LH. Duodenitis-proximal jejunitis. Equine Internal Medicine 3rd edn, 846–850.
- Kim S, Chen J, Cheng T, Gindulyte A, He J, He S, Li Q, Shoemaker BA, Thiessen PA, Yu B, Zaslavsky L, Zhang J, Bolton EE. PubChem 2019 update: improved access to chemical data.. Nucleic Acids Res 2019 Jan 8;47(D1):D1102-D1109.
- Kita T, Smith CE, Fok KF, Duffin KL, Moore WM, Karabatsos PJ, Kachur JF, Hamra FK, Pidhorodeckyj NV, Forte LR. Characterization of human uroguanylin: a member of the guanylin peptide family.. Am J Physiol 1994 Feb;266(2 Pt 2):F342-8.
- Kloeters O, Friess H, Giese N, Buechler MW, Cetin Y, Kulaksiz H. Uroguanylin inhibits proliferation of pancreatic cancer cells.. Scand J Gastroenterol 2008;43(4):447-55.
- Krogh A, Larsson B, von Heijne G, Sonnhammer EL. Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes.. J Mol Biol 2001 Jan 19;305(3):567-80.
- Laskowski RA, MacArthur MW, Thornton JM. PROCHECK: validation of protein structure coordinates. International Tables of Crystallography, Volume F. Crystallography of Biological Macromolecules pp. 722–725.
- Laus F, Gialletti R, Cappelli K, Fruganti A, Marini C, Marchegiani A. Expression of guanylin, uroguanylin and guanylate cyclase-c receptors on different gastrointestinal tracts of horse. Proceedings of the 72nd Congress Sisvet, Nizza .
- Laus F, Paggi E, Cerquetella M, Spaziante D, Spaterna A, Tesei B. Guttural pouch mycosis in a donkey (Equus asinus): a case report. Vet. Med. 55 561–565.
- Laus F, Fratini M, Paggi E, Faillace V, Spaterna A, Tesei B, Fettucciari K, Bassotti G. Effects of Single-Dose Prucalopride on Intestinal Hypomotility in Horses: Preliminary Observations.. Sci Rep 2017 Jan 27;7:41526.
- Lembo AJ, Schneier HA, Shiff SJ, Kurtz CB, MacDougall JE, Jia XD, Shao JZ, Lavins BJ, Currie MG, Fitch DA, Jeglinski BI, Eng P, Fox SM, Johnston JM. Two randomized trials of linaclotide for chronic constipation.. N Engl J Med 2011 Aug 11;365(6):527-36.
- Lin JE, Valentino M, Marszalowicz G, Magee MS, Li P, Snook AE, Stoecker BA, Chang C, Waldman SA. Bacterial heat-stable enterotoxins: translation of pathogenic peptides into novel targeted diagnostics and therapeutics.. Toxins (Basel) 2010 Aug;2(8):2028-54.
- Lucas KA, Pitari GM, Kazerounian S, Ruiz-Stewart I, Park J, Schulz S, Chepenik KP, Waldman SA. Guanylyl cyclases and signaling by cyclic GMP.. Pharmacol Rev 2000 Sep;52(3):375-414.
- Mashiach E, Schneidman-Duhovny D, Andrusier N, Nussinov R, Wolfson HJ. FireDock: a web server for fast interaction refinement in molecular docking.. Nucleic Acids Res 2008 Jul 1;36(Web Server issue):W229-32.
- Miner PB Jr. Efficacy and safety of plecanatide in treating constipation predominant irritable bowel syndrome.. Expert Opin Pharmacother 2018 Feb;19(2):177-183.
- Nieto JE, Maher O, Stanley SD, Larson R, Snyder JR. In vivo and in vitro evaluation of the effects of domperidone on the gastrointestinal tract of healthy horses.. Am J Vet Res 2013 Aug;74(8):1103-10.
- Quigley EM. Cisapride: what can we learn from the rise and fall of a prokinetic?. J Dig Dis 2011 Jun;12(3):147-56.
- Salem SE, Proudman CJ, Archer DC. Prevention of post operative complications following surgical treatment of equine colic: Current evidence.. Equine Vet J 2016 Mar;48(2):143-51.
- Sanchez LC. Gastrointestinal ileus. Equine Internal Medicine 3rd edn, 802–807.
- Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis.. Nat Methods 2012 Jul;9(7):671-5.
- Schneidman-Duhovny D, Inbar Y, Nussinov R, Wolfson HJ. PatchDock and SymmDock: servers for rigid and symmetric docking.. Nucleic Acids Res 2005 Jul 1;33(Web Server issue):W363-7.
- Shailubhai K, Palejwala V, Arjunan KP, Saykhedkar S, Nefsky B, Foss JA, Comiskey S, Jacob GS, Plevy SE. Plecanatide and dolcanatide, novel guanylate cyclase-C agonists, ameliorate gastrointestinal inflammation in experimental models of murine colitis.. World J Gastrointest Pharmacol Ther 2015 Nov 6;6(4):213-22.
- Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Söding J, Thompson JD, Higgins DG. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega.. Mol Syst Biol 2011 Oct 11;7:539.
- Stefanetti V, Marenzoni ML, Passamonti F, Cappelli K, Garcia-Etxebarria K, Coletti M, Capomaccio S. High Expression of Endogenous Retroviral Envelope Gene in the Equine Fetal Part of the Placenta.. PLoS One 2016;11(5):e0155603.
- Steinbrecher KA, Cohen MB. Transmembrane guanylate cyclase in intestinal pathophysiology.. Curr Opin Gastroenterol 2011 Mar;27(2):139-45.
- . UniProt: a worldwide hub of protein knowledge.. Nucleic Acids Res 2019 Jan 8;47(D1):D506-D515.
- Uranga JA, Castro M, Abalo R. Guanylate Cyclase C: A Current Hot Target, from Physiology to Pathology.. Curr Med Chem 2018;25(16):1879-1908.
- Van Gunsteren WF. Biomolecular Simulation: The GROMOS 96 Manual and User Guide. Zürich: VDF Hochschulverlag AG an der ETH Zürich, 1–1042.
- Van Hoogmoed LM, Nieto JE, Snyder JR, Harmon FA. Survey of prokinetic use in horses with gastrointestinal injury.. Vet Surg 2004 May-Jun;33(3):279-85.
- Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes.. Genome Biol 2002 Jun 18;3(7):RESEARCH0034.
- Waldman SA, Camilleri M. Guanylate cyclase-C as a therapeutic target in gastrointestinal disorders.. Gut 2018 Aug;67(8):1543-1552.
- Whitaker TL, Witte DP, Scott MC, Cohen MB. Uroguanylin and guanylin: distinct but overlapping patterns of messenger RNA expression in mouse intestine.. Gastroenterology 1997 Sep;113(3):1000-6.
- Wiegand RC, Kato J, Huang MD, Fok KF, Kachur JF, Currie MG. Human guanylin: cDNA isolation, structure, and activity.. FEBS Lett 1992 Oct 19;311(2):150-4.
- Xu D, Zhang Y. Improving the physical realism and structural accuracy of protein models by a two-step atomic-level energy minimization.. Biophys J 2011 Nov 16;101(10):2525-34.
- Yang J, Yan R, Roy A, Xu D, Poisson J, Zhang Y. The I-TASSER Suite: protein structure and function prediction.. Nat Methods 2015 Jan;12(1):7-8.
Citations
This article has been cited 1 times.- Pandey KN. Guanylyl cyclase/natriuretic peptide receptor-A: Identification, molecular characterization, and physiological genomics.. Front Mol Neurosci 2022;15:1076799.
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