Project description:Sexually mature (mid-May) adult female goldfish received a single injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; 50 ug/g body weight at 5 uL/g body weight) or 0.6% saline on day 0 and a single injection of alpha-methyl-p-tyrosine (aMPT; 240 ug/g body weight at 5 uL/g body weight) or 0.6% saline on Day 5 with the goal of severely deplete catecholamine (dopamine and norepinephrine) levels. Hypothalamus tissue was extracted on Day 6. Telencephalon tissue was extracted on Day 6. The objective of this study was to examine gene expression changes in the neuroendocrine brain in response to catecholamine depletion. A common reference design was used for both the hypothalamus and telencephalon groups: Hypothalamus: 3 independent groups of hypothalami from sexually mature female goldfish treated with 50 ug/g MPTP + 240 ug/g aMPT hybridized against a common vehicle control. An additional group was used for a dye-swap. Slides 4, 17, 21, 52. Telencephalon: 3 independent groups of telencephali from sexually mature female goldfish treated with 50 ug/g MPTP + 240 ug/g aMPT hybridized against a common vehicle control. An additional group was used for a dye-swap. Slides 28, 34, 43, 48.
Project description:Sexually inactive male goldfish were injected i.p. with lumiestrone (0.5 µg/g) or control vehicle (in charcoal-stripped peanut oil; 5 µL/g) to investigate the biological activity of lumiestrone. A common reference design was used: 4 independent groups of sexually inactive male goldfish treated with 0.5 ug/g lumiestrone hybridized against a common vehicle control. Slides 2, 15, 20, 43.
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
Project description:Sexually mature female goldfish were administered 2 i.p. injections: 1) 0.1% DMSO in 0.6% saline or SKF 38393 (40 µg/g) dissolved in 0.1% DMSO in 0.6% saline, and; 2) 0.6% saline. A common reference design was used: 4 independent groups of hypothalami from sexually mature female goldfish treated with 40 ug/g SKF 38393 hybridized against a common vehicle control. Slides 9, 19, 33, 44.
Project description:Sexually mature female goldfish were administered 2 i.p. injections: 1) 0.1% DMSO in 0.6% saline or SKF 38393 (40 µg/g) dissolved in 0.1% DMSO in 0.6% saline, and; 2) 0.6% saline. A common reference design was used: 4 independent groups of telencephali from sexually mature female goldfish treated with 40 ug/g SKF 38393 hybridized against a common vehicle control. Slides 7, 13, 30, 47.
Project description:Sexually mature female goldfish were administered 2 i.p. injections: 1) 0.1% DMSO in 0.6% saline, and; 2) 0.6% saline OR quinpirole (2 ug/g) dissolved in 0.6% saline. A common reference design was used: 4 independent groups of telencephali from sexually mature female goldfish treated with 2 ug/g quinpirole hybridized against a common vehicle control. Slides 8, 16, 24, 40.
Project description:Sexually regressing female goldfish were administered 2 i.p. injections: 1) 40 ug/g SCH 23390 in 0.1% DMSO and 0.6% saline, and; 2) 0.6% saline Tissues were harvested 5h post-injection. A common reference design was used: 4 independent groups of telencephali from sexually regressing female goldfish treated with 40 ug/g SCH 23390 hybridized against a common vehicle control. Slides 16, 24, 30, 35
Project description:Sexually regressing female goldfish were administered 2 i.p. injections: 1) 6 ug/g sulpiride in 0.1% DMSO and 0.6% saline, and; 2) 0.6% saline Tissues were harvested 5h post-injection. A common reference design was used: 4 independent groups of telencephali from sexually regressing female goldfish treated with 6 ug/g sulpiride hybridized against a common vehicle control. Slides 12, 26, 37, 48
Project description:Sexually regressing female goldfish were administered 2 i.p. injections: 1) 6 ug/g sulpiride in 0.1% DMSO and 0.6% saline, and; 2) 0.6% saline Tissues were harvested 5h post-injection. A common reference design was used: 4 independent groups of hypothalami from sexually mature female goldfish treated with 6 ug/g sulpiride hybridized against a common vehicle control. Slides 6, 15, 28, 44.
Project description:Sexually regressing female goldfish were administered 2 i.p. injections: 1) 40 ug/g SCH 23390 in 0.1% DMSO and 0.6% saline, and; 2) 0.6% saline Tissues were harvested 5h post-injection. A common reference design was used: 4 independent groups of telencephali from sexually regressing female goldfish treated with 40 ug/g SCH 23390 hybridized against a common vehicle control. Slides 3, 19, 36, 41