GEO Accession viewer

NCBI > GEO > Accession Display

Not logged in | Login

GEO help: Mouse over screen elements for information.

Series GSE7018

Query DataSets for GSE7018

Status

Public on Dec 29, 2007

Title

Androgen treatment in female rainbow trout leads to a marked dysregulation of gonadal gene expression profiles

Organism

Oncorhynchus mykiss

Experiment type

Expression profiling by array

Summary

The rainbow trout, Oncorhynchus mykiss, has a male heterogametic XY genetic system, and this knowledge can be used to produce experimentally all male or all female genetic populations using males with new genotypes (XX and YY males). These monosex populations have been widely used for sex differentiation studies because they give the opportunity to work on undifferentiated gonads for which the natural fate as testis or ovary is known a priori. Using as a resource the availability of a lot of expressed sequenced tags (ESTs) sequencing projects in trout, we designed and built a micro-array in order to characterize, at the pangenomic scale, rainbow trout natural gonadal differentiation as well as the mechanisms by which androgen masculinize the embryonic ovary. We choose a Nylon membrane array technique used for large-scale gene expression profiling with low cost, easy customization and high sensitivity, which is important when a limiting amount of RNA is available.
Keywords: time course of natural and androgen induced gonadal sex differentiation

Overall design

Animals and samplings : all-male (male control group ‘Male’) and all-female (female control group ‘Female’) rainbow trout populations were obtained at the INRA experimental fish farm (Sizun, France) as previously described [10471475] Treatment with androgens (female treated Group ‘Female11ß’) was carried out as previously described [11748604]. In each group, 20 to 100 gonads, according to the age of the fish, were sampled and pooled in duplicates at various stages of development [16014816]: onset of the free swimming period after complete yolk resumption (Day 0 = D0), D0+7days (D7), occurrence of oocyte meiosis (D12), beginning of ovarian lamellar structures development (D27), occurrence of previtellogenic oocytes (D60), D90 and D110. They were immediately frozen in liquid nitrogen and stored at -80°C until RNA extraction.
Total RNA extraction : total RNA was extracted using TRIzol reagent (Invitrogen, Cergy Pontoise, France) as previously described [11316749]. The total RNA concentration was determined with an Agilent 2100 Bioanalyzer and the RNA 6000 LabChip® kit (Agilent Technologies, Stockport, UK) according to the manufacturers’ instructions.
DNA microarray hybridizations : microarrays were hybridized with two types of 33P-labeled probes. The first one was an oligonucleotide having a common sequence to all spotted PCR-products (called vector hybridization) in order to determine the amount of target DNA accessible to hybridization in each spot. After stripping, a second hybridization was performed with complex probes made from 1 μg of retrotranscribed total RNA [10441335; 10445855; 11971868]. Protocol descriptions for probes preparation, hybridizations and washes are available at http://tagc.univ-mrs.fr/oncogenomics/Nylon_microarrays.php. After stringent washes, arrays were exposed to phosphor-imaging plates that were scanned with a FUJI BAS 5000 at 25 μm resolution. Hybridization signals were quantified using ArrayGauge software (Fuji Ltd, Tokyo, Japan). Signal intensities were corrected for the amount of spotted DNA and the variability of experimental conditions as previously described [10441335; 10445855].

Contributor(s)

Baron D, Montfort J, Houlgatte R, Fostier A, Guiguen Y

Citation(s)

17916255

Submission date

Feb 13, 2007

Last update date

Mar 16, 2012

Contact name

Daniel BARON

E-mail(s)

daniel.baron@univ-nantes.fr