D5_6
Protocols: The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity.
ENA first public
2021-05-16
ENA last update
2021-05-16
External Id
SAMEA6869905
INSDC center alias
College of Ocean and Earth Sciences, Xiamen University; Third Institute of Oceanography, Ministry of Natural Resources
INSDC center name
College of Ocean and Earth Sciences, Xiamen University; Third Institute of Oceanography, Ministry of Natural Resources
INSDC first public
2021-05-16T00:08:59Z
INSDC last update
2021-05-16T00:08:59Z
INSDC status
public
Submitter Id
E-MTAB-9144:D5_6
age
19
broker name
ArrayExpress
common name
Indian medaka
developmental stage
embryo
organism part
whole organism
sample name
E-MTAB-9144:D5_6