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| Effect of IP3R2 Domains on Ca2+ Secretion of Sansui Duck (Anas platyrhyncha domestica) Endometrial Epithelial Cells |
| YANG Suan, TAN Guang-Hui, ZHANG Yi-Yu*, YOU Min-Fang, LIAO Chao-Mei, TAN Yuan-Cheng, WANG Ying-Tong |
| Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education/College of Animal Science, Guizhou University, Guiyang 550025, China |
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Abstract The inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) is an intracellular Ca2+-release channel located on the endoplasmic reticulum (ER), which can regulate Ca2+ release and affect eggshell quality. To elucidate the regulatory effect of IP3R2 protein domain on Ca2+ secretion of duck (Anas platyrhyncha domestica) endometrial epithelial cells and its effect on calcium transport-related genes, in this study, the IP3R2 protein domain expression vector PCDNA3.1-IP3R2 was constructed in laying duck (Sansui duck) and transfected into duck endometrial epithelial cells. The expression of IP3R2 protein domain was detected by Flag label detection kit (ELISA); Fluo-3/AM calcium fluorescence label was used to detect intracellular Ca2+ content; qPCR was used to detect the overexpression levels of different domains and mRNA levels of ion transport-related genes. The results showed that overexpression of MIR, RYDR_ITPR (1), RYDR_ITPR(2), RIH_assoc and Ion_trans domains all caused the decrease of Ca2+ concentration in duck endometrial epithelial cells; The expression levels of RYDR_ITPR(1) and IP3R2-Ion_trans were significantly negatively correlated with intracellular Ca2+ concentration (P<0.05); The expression levels of RYDR_ITPR(2), RIH_assoc and 5 domains were significantly negatively correlated with intracellular Ca2+ concentration (P<0.01); The expression of RYDR_ITPR(2) domain was significantly positively correlated with the expression of RYR2, KCNJ15 and CA2 genes (P<0.05); The overexpression levels of MIR-domain were significantly positively correlated with the gene expression levels of IP3R1, CALB1 and CA2 (P<0.05), and the overexpression levels of the 2 RYDR_ITPR domains were significantly positively correlated with the gene expression levels of KCNJ15 and RYR2 (P<0.05); The overexpression of RIH_assoc domain was significantly positively correlated with the expression of IP3R2 and ORAI1 genes (P<0.05); and the overexpression of Ion_trans domain was significantly positively correlated with the expression of CALB1, SLC8A1 and CA2 genes (P<0.05). The co-transfection level of the five domains were significantly positively correlated with CFTR gene expression, and negatively correlated with TRPV6 gene expression (P<0.05). These results suggest that overexpression of 5 IP3R2 protein domains can cause extracellular Ca2+ release in duck endometrial epithelial cells, and have different regulatory effects on the expression levels of 20 ion transport genes. This study provides a reference for improving eggshell quality of ducks in terms of molecular genetic marker selection.
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Received: 23 August 2021
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Corresponding Authors:
* zyy8yyc@163.com
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