158 The Identification of Pig Calcium-sensing Receptor (pCaSR) Expression in Porcine Intestinal Enterocytes

Enterocytes play important roles in nutrient absorption, while the intestinal porcine enterocyte cell line (IPEC-J2) is a non-transformed and permanent commercial cell line. The calcium-sensing receptor (CaSR) has been identified as calcium ions and L-amino acids sensor in the gut and studies have demonstrated that the CaSR is involved in nutrient digestion and absorption, as well as gut barrier function. Although the expression of CaSR on the basal membrane of the villus has been found in other animal species, its expression in porcine enterocytes (pCaSR) has not been investigated to date. To investigate the expression of pCaSR in isolated porcine enterocytes and IPEC-J2, cell sorting of ileal enterocytes by fluorescence-activated cell sorting (FACS) based on sucrase-isomaltose as enterocytes marker were performed to obtain pure porcine enterocytes. The digital droplet PCR (ddPCR), immunofluorescence staining, and Western blotting were applied for the detection of pCaSR expression at the gene and protein levels, respectively. Our results showed that about 3.3% of upper epithelial cells were characterized and sorted as pure porcine enterocytes, while the other cells were negative to the enterocytes marker. In addition, neither the isolated porcine enterocytes nor the IPEC-J2 expressed the pCaSR. In summary, pure porcine enterocytes could be obtained by using FACS with the sucrase-isomaltase as enterocytes marker but pCaSR is not expressed in either isolated porcine enterocytes or IPEC-J2, which provides new insights for future work exploring the role of pCaSR in the intestine.