RNA interference-mediated knockdown of the mouse gene encoding potassium channel subfamily K member 10 inhibits hormone-induced differentiation of 3T3-L1 preadipocytes.
Intramuscular fat (IMF) is an economically important trait of domestic meat animals; thus, it is important to identify the factors that influence the IMF content. In this study, we identified the gene associated with adipogenesis from all the positional candidate genes located in the quantitative trait loci (QTL) for IMF content on porcine chromosome 7. We analyzed the expression of the abovementioned genes during differentiation of mouse 3T3-L1 preadipocytes by using real-time polymerase chain reaction (PCR). Total cellular RNA was extracted before and 6, 12, 36, and 48 h and 4, 6, and 8d after treatment with standard hormonal inducers of differentiation-insulin, dexamethasone, and 3-isobutyl-1-methylxanthine (IBMX). Six hours after induction, potassium channel subfamily K member 10 (KCNK10) gene expression in the preadipocytes was found to be 100-fold greater than that at the baseline; this expression declined until day 4 after the induction. Moreover, knockdown of the KCNK10 gene by transfection with short-hairpin RNA (shRNA) significantly decreased triacylglycerol accumulation on day 8 after the induction. An RNA interference study revealed that KCNK10 knockdown inhibited the differentiation of 3T3-L1 cells. These results indicate that KCNK10 plays an important role in the early stages of preadipocyte differentiation.
Sato
Shuji
S
National Livestock Breeding Center, Nishigo, Fukushima 961-8511, Japan. s0sato@nlbc.go.jp
Nishizuka
Makoto
M
Asano
Mami
M
Ohtake
Tsuyoshi
T
Imagawa
Masayoshi
M
Kobayashi
Eiji
E
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology* / Q000187:drug effects; D050156:Adipogenesis / Q000187:drug effects / Q000235:genetics; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D015536:Down-Regulation; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D006728:Hormones / Q000494:pharmacology*; D051379:Mice; D016133:Polymerase Chain Reaction; D024683:Potassium Channels, Tandem Pore Domain / Q000172:deficiency* / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D013997:Time Factors
D006728:Hormones; C503445:Kcnk10 protein, mouse; D024683:Potassium Channels, Tandem Pore Domain; D012333:RNA, Messenger; D034741:RNA, Small Interfering