|Application:||Gene expression microarray analysis|
|Number of samples:||59|
|Release date:||Jan 15 2016|
|Last update date:||Apr 1 2016|
|Dataset link||Oyster reproduction is affected by exposure to polystyrene microplastics|
To assess the impact of polystyrene microspheres (PS) on the physiology of the Pacific oyster, adult oysters were experimentally exposed to virgin micro-PS (2 and 6 µm in diameter; 32 µg L-1) for two months during a reproductive cycle and compared to control oysters. Adults were sampled 2 and 8 weeks after the beginning of exposure (corresponding to T1 and T3, respectively, 8-9 replicates per time of sampling and condition for a total of 56). Tissues were immediately dissected from each oyster, frozen in liquid nitrogen, then crushed to a fine powder at -196°C with an oscillating mill mixer and stored in liquid nitrogen until RNA extraction. Oocytes were collected from 5 females per condition, filtered in a 40 µm sieve, counted and transferred into 1.5 mL of Extract-all reagent (Eurobio, Courtaboeuf, France) (20,000 oocytes). Total RNA was isolated using 1.5 mL of Extract-all Reagent per 50 mg of gonad powder. For microarray hybridizations, 200 ng of total RNA were indirectly labeled with Cy3 using the Low Input Quick Amp Labeling kit One-Color. Hybridization was performed using the Agilent Gene expression hybridization kit (5188-5242), with 1.65 μg of labeled RNA, for 16 h at 65 °C. The employed arrays were Agilent 60-mer 4x44K custom microarrays, containing 31,918 C. gigas ESTs, designed by Dheilly et al. (2011). Slides were scanned on an Agilent Technologies G2565AA Microarray Scanner system at 5 μm resolution, using default parameters. Features were extracted using the Agilent Feature Extraction software 6.1.