norven

Published: 01-11-2020| Version 1 | DOI: 10.17632/svr9kvsngy.1
Contributors:
Pedro Rodrigues,
luis Oliva teles,
Laura Guimaraes

Description

The data presented herein relates to the article entitled "Effects of norfluoxetine and venlafaxine in zebrafish larvae: molecular data" by Rodrigues et al., submitted to Data In Brief. The file contains raw data corresponding to expression of several zebrafish genes related to mode of action of psychoactive compounds acting upon serotonergic or/and noradrenergic neurotransmission. Zebrafish embryos were exposed to concentrations of norfluoxetine (0.64 to 400 ng/L), venlafaxine (16 to 10000 ng/L) or their combination (3.2 ng/L norfluoxetine + 2000 ng/L venlafaxine) over 80 hpf (hours post-fertilisation). At the end of the exposure, larvae were collected for quantification of the expression of 34 genes. Gene expression was evaluated by qPCR (SYBRGreen). Details on exposure conditions, gene sequences, primers employed and assay conditions are provided in the submitted article. The work was supported by funding EU and FCT/UEFISCDI/FORMAS funding through project REWATER (ERA-NET Cofund WaterWorks2015, Water JPI) and by national funds through FCT (Portuguese Foundation for the Science and Technology) within the scope of UIDB/04423/2020 and UIDP/04423/2020. PR was supported by a PhD fellowship (SFRH/BD/134518/2017) from FCT.

Files

Steps to reproduce

All protocols and details are provided in Rodrigues et al. submitted to Data In Brief. Briefly, RNA was extracted using the Illustra RNAspin Mini RNA Isolation kit (GE Healthcare). RNA quality was checked by electrophoresis on an agarose gel of the 18s band and by measuring the optical density ratio at λ260/280nm in a BioTek spectrophotometer. Then, 1µg of total RNA was subjected to digestion of genomic DNA using deoxyribonuclease I Amplification Grade (Invitrogen) and cDNA synthesis was performed using iScript cDNA Synthesis Kit (Biorad). Design of primer pairs was based on gene sequences available in public databases using Primer 3 Plus program. Quantitative real time PCR (qRT-PCR) was employed to assess gene expression. The highest fluorescence signal reached for the lower Cycle threshold (Ct) was used to dictate ideal primer concentrations for qRT-PCR. Primer efficiency was assessed by a series of eight cDNA dilutions ranging from 0.05 to 50ng/μL. The qRT-PCR reactions (10µL of SybrGreen (Biorad), 4µL of water, 2µL of forward primer, 2µl of reverse primer and 2µL of cDNA, in a 20 µL reaction volume) were run in an Eppendorf Mastercycler realplex 4. The reaction parameters were set as follows: 94ºC for 2min; 40 cycles for 30sec at 94ºC for denaturation, for 30sec at respective annealing temperatures, and for another 30 sec at 72ºC for extension; a final extension cycle of 10 minutes at 72ºC was applied. Annealing temperatures were 51ºC for vmat2, 55ºC for 5-ht2c and drdb1, 54ºC for the remaining genes. Blank samples, as well as, melting curves were run for each of the genes assessed (Rodrigues et al. Submitted). Rodrigues P, Cunha V, Oliva-Teles L, Ferreira M, Guimarães L. Effects of norfluoxetine and venlafaxine in zebrafish larvae: molecular data. Data In Brief, submitted