title: Establishment of a versatile cell line for juvenile hormone signaling analysis in Tribolium castaneum.
abstract: The red flour beetle, Tribolium castaneum, has been widely used as a laboratory model for analyzing gene function. In this study, we established a novel cell line (Tc81) from T. castaneum embryos and validated the utility of this cell line by analyzing the juvenile hormone (JH) signaling pathway. In Tc81 cells, the Krüppel homolog 1 gene (Kr-h1), which is a JH-dependent repressor of insect metamorphosis, was rapidly induced by subnanomolar levels of JHs. Bioinformatics analysis and reporter assays identified 2 JH response elements (kJHREs) located in the region upstream of the transcription start site and in the first intron of Kr-h1. Furthermore, methoprene tolerant (Met) and steroid receptor co-activator (SRC) RNAi reduced JH-dependent induction of Kr-h1 transcripts and kJHRE-reporter activities. Thus, this novel Tc81 cell line is useful for the elucidation of JH signaling and is a promising tool for the functional analysis of genes by RNAi and reporter assays.
authors.affiliation: Division of Insect Sciences, National Institute of Agrobiological Sciences, Ohwashi 1-2, Tsukuba, Ibaraki 305-8634, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Kayukawa
authors.forename: Ken
authors.initials: K
authors.lastname: Tateishi
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Shinoda
mesh_terms: D000818:Animals; D002460:Cell Line; D004330:Drosophila; D004351:Drug Resistance / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D007605:Juvenile Hormones / Q000235:genetics / Q000378:metabolism*; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008726:Methoprene / Q000378:metabolism / Q000494:pharmacology; D056920:Nuclear Receptor Coactivator 1 / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering; D020218:Response Elements / Q000235:genetics; D015398:Signal Transduction; D014231:Tribolium / Q000235:genetics / Q000378:metabolism*
chemical_list: D019476:Insect Proteins; D007605:Juvenile Hormones; D051741:Kruppel-Like Transcription Factors; D034741:RNA, Small Interfering; D008726:Methoprene; D056920:Nuclear Receptor Coactivator 1
references.citation: Proc Natl Acad Sci U S A. 2011 Dec 27;108(52):21128-33
references.pmid: 22167806
references.citation: Proc Natl Acad Sci U S A. 2012 Oct 9;109(41):16576-81
references.pmid: 23012454
references.citation: Sci Rep. 2011;1:163
references.pmid: 22355678
references.citation: Dev Biol. 2000 May 1;221(1):53-67
references.pmid: 10772791
references.citation: Mech Dev. 2008 Jul;125(7):601-16
references.pmid: 18450431
references.citation: Dev Genes Evol. 2004 Nov;214(11):575-8
references.pmid: 15365833
references.citation: In Vitro Cell Dev Biol Anim. 2001 Jun;37(6):330-7
references.pmid: 11515963
references.citation: J Chem Ecol. 1993 Apr;19(4):723-34
references.pmid: 24249013
references.citation: J Biol Chem. 2010 Nov 19;285(47):36933-44
references.pmid: 20829361
references.citation: Annu Rev Entomol. 2013;58:181-204
references.pmid: 22994547
references.citation: J Insect Physiol. 2008 Jun;54(6):895-901
references.pmid: 18355835
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 19;104(25):10488-93
references.pmid: 17537916
references.citation: Proc Natl Acad Sci U S A. 2011 Jan 11;108(2):638-43
references.pmid: 21187375
references.citation: Insect Mol Biol. 2007 Jun;16(3):265-75
references.pmid: 17316329
references.citation: Dev Biol. 2009 Jan 15;325(2):341-50
references.pmid: 19013451
references.citation: Dev Biol. 1986 Nov;118(1):190-201
references.pmid: 3095161
references.citation: Nature. 2008 Apr 24;452(7190):949-55
references.pmid: 18362917
references.citation: PLoS One. 2011;6(12):e28728
references.pmid: 22174880
references.citation: Insect Biochem Mol Biol. 2010 Jan;40(1):23-9
references.pmid: 20018242
references.citation: J Biol Chem. 2011 Mar 11;286(10):8437-8447
references.pmid: 21190938
references.citation: Insect Biochem Mol Biol. 2000 Aug-Sep;30(8-9):617-44
references.pmid: 10876106
references.citation: FEBS J. 2005 Mar;272(5):1169-78
references.pmid: 15720391
references.citation: Proc Natl Acad Sci U S A. 2012 Jul 17;109(29):11729-34
references.pmid: 22753472
references.citation: In Vitro Cell Dev Biol Anim. 2012 Aug;48(7):426-33
references.pmid: 22752637
references.citation: Gene. 2009 Dec 1;448(1):74-87
references.pmid: 19683038
references.citation: In Vitro Cell Dev Biol Anim. 2009 Mar-Apr;45(3-4):93-105
references.pmid: 19247722
references.citation: BMC Evol Biol. 2010 Apr 30;10:120
references.pmid: 20429952
references.citation: Mech Dev. 2008 Jan-Feb;125(1-2):91-105
references.pmid: 18036785
references.citation: Insect Biochem Mol Biol. 2011 Feb;41(2):125-34
references.pmid: 21111817
references.citation: Arch Insect Biochem Physiol. 1996;32(3-4):271-86
references.pmid: 8756300
references.citation: Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2761-6
references.pmid: 9501163