Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos
The aim of the study was to investigate whether phosphorus (P) transporters, type IIb sodium-dependent phosphate cotransporter (NaP-IIb) and inorganic phosphate transporter 2 (PiT2), were directly involved in P absorption across primary cultured duodenal epithelial cell monolayers of chick embryos....
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
KeAi Communications Co., Ltd.
2022-07-01
|
| Series: | Journal of Integrative Agriculture |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311921637711 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849248943045607424 |
|---|---|
| author | Ting-ting LI Na LU Yu-xin SHAO Li-yang ZHANG Lin LU Zong-ping LIU Xu-gang LUO Xiu-dong LIAO |
| author_facet | Ting-ting LI Na LU Yu-xin SHAO Li-yang ZHANG Lin LU Zong-ping LIU Xu-gang LUO Xiu-dong LIAO |
| author_sort | Ting-ting LI |
| collection | DOAJ |
| description | The aim of the study was to investigate whether phosphorus (P) transporters, type IIb sodium-dependent phosphate cotransporter (NaP-IIb) and inorganic phosphate transporter 2 (PiT2), were directly involved in P absorption across primary cultured duodenal epithelial cell monolayers of chick embryos. The siRNAs against NaP-IIb or PiT2 were designed, synthesized and transfected into primary cultured duodenal epithelial cells of chick embryos. Then, the inhibitory efficiency of siRNAs against NaP-IIb or PiT2 was analyzed, and the most efficacious siRNAs were selected to be used for subsequent P absorption experiments. Briefly, primary cultured duodenal epithelial cells of chick embryos were transfected with either NaP-IIb or PiT2 siRNAs and grown in confluent monolayers on transwell plates. The untransfected or transfected cell monolayers were then incubated in an uptake medium containing 0 or 0.25 mmol L–1 of P as KH2PO4 to measure the P absorption across duodenal epithelial cell monolayers. The results showed that among the siRNAs designed, si-1372 and si-890 were demonstrated to be the most effective in inhibiting the NaP-IIb and PiT2 expressions, respectively. Supplemental P increased (P=0.065) the protein abundance of PiT2 and enhanced (P<0.0001) P absorption in primary cultured duodenal epithelial cell of chick embryos. Furthermore, NaP-IIb silencing decreased (P=0.07) P absorption across duodenal epithelial cell monolayers, while PiT2 silencing had no effect (P=0.345). It is concluded that the NaP-IIb, but not PiT2, might be directly involved in the P absorption of chick duodenal epithelial cells. |
| format | Article |
| id | doaj-art-bd4f319f5f144189b9ccbb64764ee6d2 |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2022-07-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-bd4f319f5f144189b9ccbb64764ee6d22025-08-20T03:57:43ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192022-07-012172076208510.1016/S2095-3119(21)63771-1Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryosTing-ting LI0Na LU1Yu-xin SHAO2Li-yang ZHANG3Lin LU4Zong-ping LIU5Xu-gang LUO6Xiu-dong LIAO7Poultry Mineral Nutrition Laboratory, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, P.R.ChinaMineral Nutrition Research Division, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.ChinaMineral Nutrition Research Division, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.ChinaMineral Nutrition Research Division, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.ChinaMineral Nutrition Research Division, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou 225009, P.R.ChinaPoultry Mineral Nutrition Laboratory, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, P.R.China; Correspondence LUO Xu-gang, Tel: +86-514-87976732Mineral Nutrition Research Division, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China; Correspondence LIAO Xiu-dong, Tel: +86-10-62820866The aim of the study was to investigate whether phosphorus (P) transporters, type IIb sodium-dependent phosphate cotransporter (NaP-IIb) and inorganic phosphate transporter 2 (PiT2), were directly involved in P absorption across primary cultured duodenal epithelial cell monolayers of chick embryos. The siRNAs against NaP-IIb or PiT2 were designed, synthesized and transfected into primary cultured duodenal epithelial cells of chick embryos. Then, the inhibitory efficiency of siRNAs against NaP-IIb or PiT2 was analyzed, and the most efficacious siRNAs were selected to be used for subsequent P absorption experiments. Briefly, primary cultured duodenal epithelial cells of chick embryos were transfected with either NaP-IIb or PiT2 siRNAs and grown in confluent monolayers on transwell plates. The untransfected or transfected cell monolayers were then incubated in an uptake medium containing 0 or 0.25 mmol L–1 of P as KH2PO4 to measure the P absorption across duodenal epithelial cell monolayers. The results showed that among the siRNAs designed, si-1372 and si-890 were demonstrated to be the most effective in inhibiting the NaP-IIb and PiT2 expressions, respectively. Supplemental P increased (P=0.065) the protein abundance of PiT2 and enhanced (P<0.0001) P absorption in primary cultured duodenal epithelial cell of chick embryos. Furthermore, NaP-IIb silencing decreased (P=0.07) P absorption across duodenal epithelial cell monolayers, while PiT2 silencing had no effect (P=0.345). It is concluded that the NaP-IIb, but not PiT2, might be directly involved in the P absorption of chick duodenal epithelial cells.http://www.sciencedirect.com/science/article/pii/S2095311921637711broilerphosphorus transporterphosphorus absorptionprimary cultured duodenal epithelial cell |
| spellingShingle | Ting-ting LI Na LU Yu-xin SHAO Li-yang ZHANG Lin LU Zong-ping LIU Xu-gang LUO Xiu-dong LIAO Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos Journal of Integrative Agriculture broiler phosphorus transporter phosphorus absorption primary cultured duodenal epithelial cell |
| title | Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos |
| title_full | Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos |
| title_fullStr | Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos |
| title_full_unstemmed | Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos |
| title_short | Effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos |
| title_sort | effect of the gene silencing of phosphorus transporters on phosphorus absorption across primary cultured duodenal epithelial cell monolayers of chick embryos |
| topic | broiler phosphorus transporter phosphorus absorption primary cultured duodenal epithelial cell |
| url | http://www.sciencedirect.com/science/article/pii/S2095311921637711 |
| work_keys_str_mv | AT tingtingli effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos AT nalu effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos AT yuxinshao effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos AT liyangzhang effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos AT linlu effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos AT zongpingliu effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos AT xugangluo effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos AT xiudongliao effectofthegenesilencingofphosphorustransportersonphosphorusabsorptionacrossprimaryculturedduodenalepithelialcellmonolayersofchickembryos |