title: Selection of reliable reference genes in Caenorhabditis elegans for analysis of nanotoxicity.
abstract: Despite rapid development and application of a wide range of manufactured metal oxide nanoparticles (NPs), the understanding of potential risks of using NPs is less completed, especially at the molecular level. The nematode Caenorhabditis elegans (C.elegans) has been emerging as an environmental model to study the molecular mechanism of environmental contaminations, using standard genetic tools such as the real-time quantitative PCR (RT-qPCR). The most important factor that may affect the accuracy of RT-qPCR is to choose appropriate genes for normalization. In this study, we selected 13 reference gene candidates (act-1, cdc-42, pmp-3, eif-3.C, actin, act-2, csq-1, Y45F10D.4, tba-1, mdh-1, ama-1, F35G12.2, and rbd-1) to test their expression stability under different doses of nano-copper oxide (CuO 0, 1, 10, and 50 µg/mL) using RT-qPCR. Four algorithms, geNorm, NormFinder, BestKeeper, and the comparative ΔCt method, were employed to evaluate these 13 candidates expressions. As a result, tba-1, Y45F10D.4 and pmp-3 were the most reliable, which may be used as reference genes in future study of nanoparticle-induced genetic response using C.elegans.
authors.affiliation: Department of Biology, East Carolina University, Greenville, North Carolina, United States of America.
authors.forename: Yanqiong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Dongliang
authors.initials: D
authors.lastname: Chen
authors.forename: Michael A
authors.initials: MA
authors.lastname: Smith
authors.forename: Baohong
authors.initials: B
authors.lastname: Zhang
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Pan
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000187:drug effects* / Q000235:genetics*; D003300:Copper / Q000633:toxicity; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D015870:Gene Expression; D017238:Genes, Helminth / Q000187:drug effects*; D053768:Metal Nanoparticles / Q000633:toxicity*; D023421:Models, Animal; D036103:Nanotechnology; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D018509:DNA, Helminth; D003300:Copper; C030973:cupric oxide