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Secretoneurin: A new hormone? Auratus: The goldfish environmental genomics project

ABSTRACT: The objectives of this study were to examine the effect of secretoneurin (SN) on pituitary gene expression. A mixture of goldfish pituitaries from both males and females were dispersed using Typsin type II and DNase II enzymes. The cells were plated in 24-well culture plates at a density of ~2.5 x 105 cells/well. After at least 12-hr plating period, cells were washed with testing medium and pre-incubated for 1 hr to stabilize basal LH secretion. Dispersed pituitary cells were treated with 10 nM gfSN for 12 hours in static incubation after which the cells were carefully washed by 1X PBS. Total RNA was isolated by utilizing RNeasy Micro Kit (QIAGEN GmbH, Hilden, Germany) following the company’s standard protocol. RNA concentrations were determined using the NanoDrop ND-100 spectrophotometer (Thermo Scientific, Wilmington, DE). cDNA synthesis, cDNA purification and microarray slide hybridization followed our validated and established microarray protocol (Martyniuk et al., 2006; Zhang et al., 2009; Popesku et al., 2010).

ORGANISM(S): Carassius auratus Cyprinus carpio

PROVIDER: GSE32629 | GEO | 2011/10/06

SECONDARY ACCESSION(S): PRJNA147129

REPOSITORIES: GEO

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Secretoneurin: A new hormone? Auratus: The goldfish environmental genomics project

Project description:The objectives of this study were to examine the effect of secretoneurin (SN) on pituitary gene expression. A mixture of goldfish pituitaries from both males and females were dispersed using Typsin type II and DNase II enzymes. The cells were plated in 24-well culture plates at a density of ~2.5 x 105 cells/well. After at least 12-hr plating period, cells were washed with testing medium and pre-incubated for 1 hr to stabilize basal LH secretion. Dispersed pituitary cells were treated with 10 nM gfSN for 12 hours in static incubation after which the cells were carefully washed by 1X PBS. Total RNA was isolated by utilizing RNeasy Micro Kit (QIAGEN GmbH, Hilden, Germany) following the company’s standard protocol. RNA concentrations were determined using the NanoDrop ND-100 spectrophotometer (Thermo Scientific, Wilmington, DE). cDNA synthesis, cDNA purification and microarray slide hybridization followed our validated and established microarray protocol (Martyniuk et al., 2006; Zhang et al., 2009; Popesku et al., 2010). 4 control and 4 treatment samples were hybridized using dual channel micrarray with cy3 and cy5

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