Calcium-sensing Receptor is not Expressed in the Absorptive Enterocytes of Weaned Piglets
Date
2022-03-16
Authors
Zhao, Xiaoya
Hui, Qianqu
Azevedo, Paula A
Nyachoti, Charles M.
O, Karmin
Yang, Chengbo
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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 as an enterocyte-specific marker. The expression of CaSR was investigated in both primary enterocytes and the intestinal porcine enterocyte cell line (IPEC-J2) by droplet digital PCR (ddPCR), immunofluorescence staining, and western blotting. Results demonstrated that porcine enterocytes could be obtained using FACS with the sucrase-isomaltase 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.
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Keywords
Calcium-sensing receptor, enterocytes, enteroendocrine cells, IPEC-J2, weaned piglets