Calcium-sensing Receptor is Not Expressed in the Absorptive Enterocytes of Weaned Piglets

Overview

Journal J Anim Sci

Date 2022 Mar 16

PMID 35294536

Authors

Xiaoya Zhao

Qianru Hui

Paula Azevedo

Charles Martin Nyachoti

Karmin O

Chengbo Yang

Affiliations

Soon will be listed here.

Abstract

The calcium-sensing receptor (CaSR) is a kokumi receptor that plays an essential role in nutrient sensing and animal physiology, growth, and development. Pig CaSR (pCaSR) was identified and characterized in the intestine. However, further research is still needed to confirm the expression of CaSR in the epithelial cells isolated from weaned piglets. In this study, primary enterocytes were isolated and characterized from the ileum of weaned piglets by the Weiser distended intestinal sac technique and fluorescence-activated cell sorting (FACS) based on sucrase-isomaltase (SI) as an enterocyte-specific marker. The expression of CaSR was investigated in both primary enterocytes and the intestinal porcine enterocyte cell line-j2 (IPEC-J2) by droplet digital polymerase chain reaction (ddPCR), immunofluorescence staining, and Western blotting. Results demonstrated that porcine enterocytes could be obtained using FACS with the SI as the enterocyte-specific marker and that pCaSR is not expressed in both porcine ileal enterocytes and IPEC-J2 cells, which specifically identified the expression of pCaSR in ileal enterocytes with sensitive and specific approaches.

References

Bullen T, Forrest S, Campbell F, Dodson A, Hershman M, Pritchard D . Characterization of epithelial cell shedding from human small intestine. Lab Invest. 2006; 86(10):1052-63. DOI: 10.1038/labinvest.3700464. View

Caroppo R, Gerbino A, Debellis L, Kifor O, Soybel D, Brown E . Asymmetrical, agonist-induced fluctuations in local extracellular [Ca(2+)] in intact polarized epithelia. EMBO J. 2001; 20(22):6316-26. PMC: 125728. DOI: 10.1093/emboj/20.22.6316. View

Hekmati M, Polak-Charcon S, Ben-Shaul Y . A morphological study of a human adenocarcinoma cell line (HT29) differentiating in culture. Similarities to intestinal embryonic development. Cell Differ Dev. 1990; 31(3):207-18. DOI: 10.1016/0922-3371(90)90133-h. View

Price A, Shamardani K, Lugo K, Deguine J, Roberts A, Lee B . A Map of Toll-like Receptor Expression in the Intestinal Epithelium Reveals Distinct Spatial, Cell Type-Specific, and Temporal Patterns. Immunity. 2018; 49(3):560-575.e6. PMC: 6152941. DOI: 10.1016/j.immuni.2018.07.016. View

Oates P, Thomas C, Morgan E . Characterization of isolated duodenal epithelial cells along a crypt-villus axis in rats fed diets with different iron content. J Gastroenterol Hepatol. 1998; 12(12):829-38. DOI: 10.1111/j.1440-1746.1997.tb00380.x. View

Gao N, Dou X, Yin T, Yang Y, Yan D, Ma Z . Tryptophan Promotes Intestinal Immune Defense through Calcium-Sensing Receptor (CaSR)-Dependent Metabolic Pathways. J Agric Food Chem. 2021; 69(45):13460-13473. DOI: 10.1021/acs.jafc.1c05820. View

Bourgine J, Billaut-Laden I, Happillon M, Lo-Guidice J, Maunoury V, Imbenotte M . Gene expression profiling of systems involved in the metabolism and the disposition of xenobiotics: comparison between human intestinal biopsy samples and colon cell lines. Drug Metab Dispos. 2012; 40(4):694-705. DOI: 10.1124/dmd.111.042465. View

Busslinger G, Weusten B, Bogte A, Begthel H, Brosens L, Clevers H . Human gastrointestinal epithelia of the esophagus, stomach, and duodenum resolved at single-cell resolution. Cell Rep. 2021; 34(10):108819. DOI: 10.1016/j.celrep.2021.108819. View

Brosnahan A, Brown D . Porcine IPEC-J2 intestinal epithelial cells in microbiological investigations. Vet Microbiol. 2011; 156(3-4):229-37. PMC: 3289732. DOI: 10.1016/j.vetmic.2011.10.017. View

10.

Sriram K, Wiley S, Moyung K, Gorr M, Salmeron C, Marucut J . Detection and Quantification of GPCR mRNA: An Assessment and Implications of Data from High-Content Methods. ACS Omega. 2019; 4(16):17048-17059. PMC: 6796235. DOI: 10.1021/acsomega.9b02811. View

11.

Insel P, Wilderman A, Zambon A, Snead A, Murray F, Aroonsakool N . G Protein-Coupled Receptor (GPCR) Expression in Native Cells: "Novel" endoGPCRs as Physiologic Regulators and Therapeutic Targets. Mol Pharmacol. 2015; 88(1):181-7. PMC: 4468643. DOI: 10.1124/mol.115.098129. View

12.

Chougule P, Herlenius G, Hernandez N, Patil P, Xu B, Sumitran-Holgersson S . Isolation and characterization of human primary enterocytes from small intestine using a novel method. Scand J Gastroenterol. 2012; 47(11):1334-43. PMC: 3490477. DOI: 10.3109/00365521.2012.708940. View

13.

Modvig I, Kuhre R, Jepsen S, Xu S, Engelstoft M, Egerod K . Amino acids differ in their capacity to stimulate GLP-1 release from the perfused rat small intestine and stimulate secretion by different sensing mechanisms. Am J Physiol Endocrinol Metab. 2021; 320(5):E874-E885. DOI: 10.1152/ajpendo.00026.2021. View

14.

Aggarwal A, Hobaus J, Tennakoon S, Prinz-Wohlgenannt M, Graca J, Price S . Active vitamin D potentiates the anti-neoplastic effects of calcium in the colon: A cross talk through the calcium-sensing receptor. J Steroid Biochem Mol Biol. 2015; 155(Pt B):231-8. DOI: 10.1016/j.jsbmb.2015.02.006. View

15.

Insel P, Snead A, Murray F, Zhang L, Yokouchi H, Katakia T . GPCR expression in tissues and cells: are the optimal receptors being used as drug targets?. Br J Pharmacol. 2011; 165(6):1613-1616. PMC: 3372817. DOI: 10.1111/j.1476-5381.2011.01434.x. View

16.

Mine Y, Zhang H . Calcium-sensing receptor (CaSR)-mediated anti-inflammatory effects of L-amino acids in intestinal epithelial cells. J Agric Food Chem. 2015; 63(45):9987-95. DOI: 10.1021/acs.jafc.5b03749. View

17.

Bolger A, Lohse M, Usadel B . Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014; 30(15):2114-20. PMC: 4103590. DOI: 10.1093/bioinformatics/btu170. View

18.

Zhao W, Zhang Z, Zheng L, You C, Chi H, Zhang T . Calcium-sensing receptor activating ERK1/2 and PI3K-Akt pathways to induce the proliferation of osteosarcoma cells. Clin Exp Pharmacol Physiol. 2019; 47(3):517-519. DOI: 10.1111/1440-1681.13209. View

19.

Farre R, Fiorani M, Abdu Rahiman S, Matteoli G . Intestinal Permeability, Inflammation and the Role of Nutrients. Nutrients. 2020; 12(4). PMC: 7231157. DOI: 10.3390/nu12041185. View

20.

Liao Y, Smyth G, Shi W . The Subread aligner: fast, accurate and scalable read mapping by seed-and-vote. Nucleic Acids Res. 2013; 41(10):e108. PMC: 3664803. DOI: 10.1093/nar/gkt214. View