title: Establishment of a cell line from the ash and privet borer beetle Tylonotus bimaculatus Haldeman and assessment of its sensitivity to diacylhydrazine insecticides.
abstract: A novel cell line, NRCAN-Tb521, was developed from larvae of the longhorn beetle Tylonotus bimaculatus (Coleoptera: Cerambycidae), a pest of North American ash trees. The cell line has been successfully passaged more than 50 times and displayed very strong attachment to the substrate and a modal chromosomal count distribution of 19. Sequencing of a 649 bp fragment of the mitochondrial cytochrome oxidase I gene confirmed the identity of NRCAN-Tb521 as T. bimaculatus. The response of the cell line to 20-hydroxyecdysone and diacylhydrazine ecdysone agonist insecticides was also studied. At 10(-6) M, 20-hydroxyecdysone, tebufenozide, methoxyfenozide and halofenozide triggered the production of numerous filamentous cytoplasmic extensions, and the cells tended to form aggregates, indicative of a cell differentiation response. This response was followed by a strong decrease in viability after 4 d. Reverse transcription polymerase chain reaction (PCR) experiments and sequencing of PCR fragments showed that the 20E receptor gene EcR is expressed in the cells and that 20E, tebufenozide, methoxyfenozide and halofenozide also induce the expression of the nuclear hormone receptor gene HR3. This report establishes that NRCAN-Tb521 is a valuable in vitro model to study effects of ecdysone agonists in wood-boring cerambycids.
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada, N1G 2W1.|Canadian Forest Service, Natural Resources Canada, 1219 Queen St. East, Sault Ste Marie, ON, Canada, P6A 2E5.
authors.forename: Fayuan
authors.initials: F
authors.lastname: Wen
authors.affiliation: Canadian Forest Service, Natural Resources Canada, 1219 Queen St. East, Sault Ste Marie, ON, Canada, P6A 2E5.
authors.forename: Guido
authors.initials: G
authors.lastname: Caputo
authors.affiliation: Canadian Forest Service, Natural Resources Canada, 1219 Queen St. East, Sault Ste Marie, ON, Canada, P6A 2E5.
authors.forename: Sharon
authors.initials: S
authors.lastname: Hooey
authors.affiliation: Canadian Forest Service, Natural Resources Canada, 1219 Queen St. East, Sault Ste Marie, ON, Canada, P6A 2E5.
authors.forename: Susan
authors.initials: S
authors.lastname: Bowman
authors.affiliation: Canadian Forest Service, Natural Resources Canada, 1219 Queen St. East, Sault Ste Marie, ON, Canada, P6A 2E5.
authors.forename: Kristine
authors.initials: K
authors.lastname: Pinkney
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada, N1G 2W1.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Krell
authors.affiliation: Canadian Forest Service, Natural Resources Canada, 1219 Queen St. East, Sault Ste Marie, ON, Canada, P6A 2E5.
authors.forename: Basil
authors.initials: B
authors.lastname: Arif
authors.affiliation: Canadian Forest Service, Natural Resources Canada, 1219 Queen St. East, Sault Ste Marie, ON, Canada, P6A 2E5. Dan.Doucet@NRCan.gc.ca.
authors.forename: Daniel
authors.identifier: http://orcid.org/0000-0003-0058-255X
authors.initials: D
authors.lastname: Doucet
mesh_terms: D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D002460:Cell Line / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D001517:Coleoptera / Q000166:cytology* / Q000187:drug effects*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004441:Ecdysterone / Q000819:agonists / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000235:genetics; D006834:Hydrazines / Q000494:pharmacology; D019476:Insect Proteins / Q000235:genetics; D007306:Insecticides / Q000494:pharmacology*; D007605:Juvenile Hormones / Q000494:pharmacology; D007621:Karyotyping; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D011987:Receptors, Steroid / Q000235:genetics
chemical_list: D001565:Benzoates; D006834:Hydrazines; D019476:Insect Proteins; D007306:Insecticides; D007605:Juvenile Hormones; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; C055469:ecdysone receptor; D004441:Ecdysterone; C406712:methoxyfenozide; C437546:N-4-chlorobenzoyl-N'-benzoyl-N'-tert-butylhydrazine; D003576:Electron Transport Complex IV; C082026:tebufenozide
keywords: Cell line; Cerambycid; Coleoptera; Ecdysone; Ecdysone agonist