Unknown,Transcriptomics,Genomics,Proteomics

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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). 4 control and 4 treatment samples were hybridized using dual channel micrarray with cy3 and cy5

ORGANISM(S): Carassius auratus

SUBMITTER: Christopher Martyniuk

PROVIDER: E-GEOD-32629 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Sex pheromones rapidly affect endocrine physiology and behaviour, but little is known about their effects on gene expression in the neuroendocrine tissues that mediate olfactory processing. In this study we exposed male goldfish for 6 h to waterborne 17,20betaP (4.3 nM) and PGF2α (3 nM), the main pre-ovulatory and post-ovulatory pheromones, respectively. Both treatments elevated milt volume (P = 0.001). Microarray hybridizations were used to male telencephalon samples following PGF2α treatment and identified 71 unique transcripts that were differentially expressed (q < 5%; 67 up, 4 down). Functional annotation of these regulated genes indicates that PGF2α pheromone exposure affects diverse biological processes including nervous system functions, energy metabolism, cholesterol/lipoprotein transport, translational regulation, transcription and chromatin remodelling, protein processing, cytoskeletal organization, and signalling. By using real-time RT-PCR, we further validated three candidate genes, ependymin-II, calmodulin-A and aldolase C, which exhibited 3 - 5 fold increase in expression following PGF2α exposure. Expression levels of some other genes that are thought to be important for reproduction were also determined using real-time RT-PCR. Expression of sGnRH was increased by PGF2α, but not 17,20betaP, whereas cGnRH expression was increased by 17,20betaP but not PGF2α. In contrast, both pheromones increase the expression of glutamate (GluR2a, NR2A) and gamma-aminobutyric acid (GABAA gamma2) receptor subunit mRNAs. This gene expression data link milt release and rapid modulation of neuronal transcription as part of the response to female sex pheromones. A total of four microarray slides were hybridized to study the effects of PGF2α on gene expression in the telencephalon. Control samples (N = 4) were pooled to form our reference or technical replicates while treatment samples (N = 4), which formed our biological replicates, were ran separately. In addition, two dye swaps were performed for our biological samples for the treatments.

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

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