title: Effects of nicotine on zebrafish: A comparative response between a newly established gill cell line and whole gills.
abstract: A novel cell line, Danio rerio gill (DrG), derived from the gill tissue of zebrafish, was established and characterized. The cells were able to grow at a wide range of temperatures from 25°C to 32°C in Leibovitz's L-15 medium. The DrG cell line consists of epithelial-like cells with a diameter of 18-22μm. The cell line was characterized by mitochondrial 12S rRNA gene. Acute toxicity tests were conducted on D. rerio by exposing them to nicotine for 96h under static conditions. In vitro cytotoxicity of nicotine was assessed in DrG cell line using multiple endpoints such as 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), Neutral Red assay, Alamar Blue assay and Coomassie Blue protein assay. Linear correlations between each in vitro cytotoxicity assay and the in vivo mortality data were highly significant. Nicotine induced intracellular reactive oxygen species generation in DrG cell line in a concentration dependent manner. DrG cell line and zebrafish exposed to nicotine significantly increased the elevation of lipid peroxidation (LPO) while depletion of reduced glutathione (GSH), manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione S-transferase (GST) and glutathione peroxidise(GPx1a) was observed. In nicotine treated fish and cells a negative correlation between reduced glutathione and LPO was observed. In addition, the production of ROS and the resulting oxidative stress resulted in increased expression of apoptosis related genes p53 and cas3.Collectively, our result suggests that nicotine has the potential to induce reactive oxygen species (ROS) production, oxidative stress and apoptosis in DrG cell line and zebrafish.
authors.affiliation: Aquatic Animal Health Laboratory (OIE Reference for WTD), PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam - 632 509, Vellore District, Tamil Nadu, India; Institute of Genomics and Integrative Biology (IGIB), New Delhi, India.
authors.forename: K S
authors.initials: KS
authors.lastname: Nathiga Nambi
authors.affiliation: Aquatic Animal Health Laboratory (OIE Reference for WTD), PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam - 632 509, Vellore District, Tamil Nadu, India.
authors.forename: S
authors.initials: S
authors.lastname: Abdul Majeed
authors.affiliation: Aquatic Animal Health Laboratory (OIE Reference for WTD), PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam - 632 509, Vellore District, Tamil Nadu, India.
authors.forename: G
authors.initials: G
authors.lastname: Taju
authors.affiliation: Institute of Genomics and Integrative Biology (IGIB), New Delhi, India.
authors.forename: Sridhar
authors.initials: S
authors.lastname: Sivasubbu
authors.affiliation: Aquatic Animal Health Laboratory (OIE Reference for WTD), PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam - 632 509, Vellore District, Tamil Nadu, India.
authors.forename: V
authors.initials: V
authors.lastname: Sarath Babu
authors.affiliation: Aquatic Animal Health Laboratory (OIE Reference for WTD), PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam - 632 509, Vellore District, Tamil Nadu, India. Electronic address: cah_sahul@hotmail.com.
authors.forename: A S
authors.initials: AS
authors.lastname: Sahul Hameed
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005731:Ganglionic Stimulants / Q000494:pharmacology / Q000633:toxicity; D005786:Gene Expression Regulation / Q000187:drug effects*; D005880:Gills / Q000166:cytology / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000235:genetics / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D009538:Nicotine / Q000494:pharmacology* / Q000633:toxicity; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D023382:Toxicity Tests, Acute / Q000379:methods; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism; D000094765:Glutathione Peroxidase GPX1
chemical_list: D005731:Ganglionic Stimulants; D017382:Reactive Oxygen Species; D029961:Zebrafish Proteins; D009538:Nicotine; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D005982:Glutathione Transferase; D005978:Glutathione; D000094765:Glutathione Peroxidase GPX1
keywords: Acute toxicity; Apoptosis; Cytotoxicity; Danio rerio gill cell line; Nicotine; Reactive oxygen species