Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

0

In vivo gene expression in granulosa cells during pig terminal follicular development.


ABSTRACT: The aim of this study is the identification of genes and gene networks involved in pig ovarian follicular development. cDNA nylon micro-arrays (2849 sequenced clones) were designed from different libraries : four subtractive suppressive hybridization libraries (generated from small versus large and small versus medium follicles) and a pig multi-tissue cDNA library. Granulosa cells were isolated from healthy follicles (small, medium or large), 24 h or 96 h after the end of a progestagen treatment. The RNA isolated from these cells was used to hybridize the micro-arrays and hybridisations were performed in duplicate. The images were quantified using Bzscan software and the data were managed with BASE software. Statistical analysis was performed using R software. Keywords: cell type comparison Granulosa cells were isolated from healthy follicles (small, medium or large). The RNA isolated from these cells was used to hybridize nylon micro-arrays. The arrays contained PCR products from 2848 pig cDNA clones (GPL3978), spotted in duplicate on two separate fields of the same membrane (last number 1 of the local ID corresponds to the left field and 2 corresponds to right field). The data have been generated from 14 RNA follicle pools (five Large, five Medium and four Small follicles pools). For each follicle pool, 2 radioactive labellings were performed (before last number of the local ID corresponds to the duplicate number). Each membrane was exposed 6, 12 or 24 hours (depending on the saturation signal) to radioisotopic-sensitive imaging plates (BAS-2025, Fujifilm, Raytest France S.A.R.L., Courbevoie, France). The imaging plates were scanned thereafter with a phosphor imaging system at a 25 µm resolution (BAS-5000, Fujifilm, Raytest France S.A.R.L., Courbevoie, France). The images were quantified using semi-automated software, BZScan (Lopez F, Rougemont J, Loriod B, Bourgeois A, Loi L, Bertucci F, Hingamp P, Houlgatte R, and Granjeaud S. Feature extraction and signal processing for nylon DNA microarrays. BMC Genomics 5: 38, 2004). Fixed circle segmentation, i.e. a grid process with a fixed spot diameter was applied. The hybridization images were quantified by the extraction of the intensity for each spot. A new version of BZsan (AGscan solfware) is available at : http://mulcyber.toulouse.inra.fr/projects/agscan/ The experimental data were managed using the free BASE software (Saal LH, Troein C, Vallon-Christersson J, Gruvberger S, Borg A, and Peterson C. BioArray Software Environment (BASE): a platform for comprehensive management and analysis of microarray data. Genome Biol 3: SOFTWARE0003, 2002.) modified by Sigenae team to accept radioactive experiments. Finally, data consisted in (14 RNA x 2 labellings x 2 fields) 14 probes, 28 hybridisations, 42 images (14 images were filtred out for background too hight).

ORGANISM(S): Sus scrofa

SUBMITTER: Agnes Bonnet 

PROVIDER: E-GEOD-5798 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

altmetric image

Publications

In vivo gene expression in granulosa cells during pig terminal follicular development.

Bonnet A A   Lê Cao K A KA   Sancristobal M M   Benne F F   Robert-Granié C C   Law-So G G   Fabre S S   Besse P P   De Billy E E   Quesnel H H   Hatey F F   Tosser-Klopp G G  

Reproduction (Cambridge, England) 20080502 2


Ovarian antral follicular development is clearly dependent on pituitary gonadotrophins FSH and LH. Although the endocrine mechanism that controls ovarian folliculogenesis leading to ovulation is quite well understood, the detailed mechanisms and molecular determinants in the different follicular compartments remain to be clarified. The aim of this study was to identify the genes differentially expressed in pig granulosa cells along the terminal ovarian follicle growth, to gain a comprehensive vi  ...[more]

Similar Datasets

2008-04-30 | E-GEOD-5797 | biostudies-arrayexpress
2010-06-24 | E-GEOD-7635 | biostudies-arrayexpress
2010-06-30 | E-GEOD-5299 | biostudies-arrayexpress
2012-12-31 | E-GEOD-23335 | biostudies-arrayexpress
2012-12-31 | E-GEOD-24273 | biostudies-arrayexpress
2012-12-31 | E-GEOD-24221 | biostudies-arrayexpress
2010-06-29 | E-GEOD-8377 | biostudies-arrayexpress
2010-07-01 | E-GEOD-4871 | biostudies-arrayexpress
2010-07-01 | E-GEOD-5928 | biostudies-arrayexpress
2010-06-24 | E-GEOD-15614 | biostudies-arrayexpress