title: Effect of transient receptor potential vanilloid 6 gene silencing on the expression of calcium transport genes in chicken osteoblasts.
abstract: Ca2+ plays a major role in the regulation of signal transduction. Transient receptor potential vanilloid 6 is a Ca2+-selective channel that serves as an important rate-limiting step in the facilitation of Ca2+ entry into cells, but little is known about the regulation of transient receptor potential vanilloid 6 in chickens. In this study, we evaluated the effects of transient receptor potential vanilloid 6 gene interference on the expression of calbindin-D28K, Na+/Ca2+ exchangers, and plasma membrane Ca2+ ATPase 1b to investigate the mechanism underlying the regulation of transient receptor potential vanilloid 6. Three hairpin siRNA expression vectors targeting transient receptor potential vanilloid 6 (pSIREN- transient receptor potential vanilloid 6) and a negative control (pSIREN-control) were constructed and transfected into chicken osteoblasts. The mRNA and protein expression levels were evaluated by quantitative reverse transcription polymerase chain reaction and Western blot, respectively. The mRNA expression levels of transient receptor potential vanilloid 6 and calbindin-D28K were reduced by 45.7% (P<0.01) and 27.9% (P<0.01), respectively, 48 h after transfection with one of the three constructs (pSIREN- transient receptor potential vanilloid 6-3) compared with the level obtained in the untreated group. There was no significant difference in the mRNA expression levels of Na+/Ca2+ exchangers and plasma membrane Ca2+ ATPase 1b. The protein expression levels of transient receptor potential vanilloid 6 and calbindin-D28K were reduced by 40.2% (P<0.01) and 29.8% (P<0.01), respectively, 48 h after transfection with pSIREN-transient receptor potential vanilloid 6-3 compared with the level obtained in the untreated group. In conclusion, the vector-based transient receptor potential vanilloid 6-shRNA can efficiently suppress the mRNA and protein expression of transient receptor potential vanilloid 6 in chicken osteoblasts, and transient receptor potential vanilloid 6 regulates the expression of calbindin-D28K during Ca2+ transport.
authors: The Bone Biological Laboratory of Livestock and Poultry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China.
authors: Jie
authors: J
authors: Zhang
authors: The Bone Biological Laboratory of Livestock and Poultry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China.
authors: Yifeng
authors: Y
authors: Deng
authors: The Bone Biological Laboratory of Livestock and Poultry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China.
authors: Huijie
authors: H
authors: Ma
authors: The Bone Biological Laboratory of Livestock and Poultry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China.
authors: Jiafa
authors: J
authors: Hou
authors: The Bone Biological Laboratory of Livestock and Poultry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, Jiangsu Province, China zhouzl@njau.edu.cn.
authors: ZhenLei
authors: Z
authors: Zhou
mesh_terms: D000818:Animals; D030161:Avian Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western / Q000662:veterinary; D064026:Calbindins / Q000235:genetics* / Q000378:metabolism; D002642:Chick Embryo; D002645:Chickens / Q000235:genetics* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D010006:Osteoblasts / Q000378:metabolism*; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000662:veterinary; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D019831:Sodium-Calcium Exchanger / Q000235:genetics / Q000378:metabolism; D050916:TRPV Cation Channels / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000662:veterinary
chemical_list: D030161:Avian Proteins; D064026:Calbindins; D034741:RNA, Small Interfering; D019831:Sodium-Calcium Exchanger; D050916:TRPV Cation Channels; D053499:Plasma Membrane Calcium-Transporting ATPases
keywords: RNAi; calbindin-D28K; chicken; osteoblasts; transient receptor potential vanilloid 6