Dataset Information

Understanding bovine germinal vesicle (GV) oocyte and cumulus cell communication... A systems biology approach

ABSTRACT: Triplicate samples of oocyte and cumulus cells were isolated from follicles of 2-8 mm, containing oocytes with several layers of cumulus cells were collected from ovaries. Proteins were extracted using DDF and digested using trypsin. The peptides were desalted, separated using in-line SCX-RPLC and analyzed using ion trap MS/MS. Proteins were identified using TurboSEQUEST 3.2

INSTRUMENT(S): instrument model, LCQ Deca XP Plus

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Cumulus Cell

SUBMITTER: Divyaswetha Peddinti

PROVIDER: PRD000082 | Pride | 2011-04-15

REPOSITORIES: Pride

ACCESS DATA

Publications

Differential detergent fractionation for non-electrophoretic eukaryote cell proteomics.

McCarthy Fiona M FM Burgess Shane C SC van den Berg Bart H J BH Koter Marek D MD Pharr G Todd GT

Journal of proteome research 20050301 2

Differential detergent fractionation (DDF), which relies on detergents to sequentially extract proteins from eukaryotic cells, has been used to increase proteome coverage of 2D-PAGE. Here, we used DDF extraction in conjunction with the nonelectrophoretic proteomics method of liquid chromatography and electrospray ionization tandem mass spectrometry. We demonstrate that DDF can be used with 2D-LC ESI MS2 for comprehensive cellular proteomics, including a large proportion of membrane proteins. Com ...[more]

PMID: 15822906

Publication: 1/4

Similar Datasets

Project description:Purpose: Preovualtory follicle diameter influences embryo quality in beef cattle that rely on a gonadotropin releasing hormone (GnRH) injection to induce the preovualtory gonadotropin surge and subsequent ovualtion. The goals of this study are to compare transcriptome profiles of pools of four oocytes or associated cumulus cells collected from small (≤11.7mm) and large follicles (≥12.7 mm) exposed to a GnRH-induced gonadotropin surge and follicles (11.7-14.0 mm) exposed to an endogenous gonadotropin surge (spontaneous follicles). Methods: Oocyte and cumulus cell mRNA profiles of pools of four oocytes or associated cumulus cells collected from small (≤11.7mm) and large follicles (≥12.7 mm) exposed to a GnRH-induced gonadotropin surge or spontaneous follicles (11.7-14.0 mm) which were exposed to an endogenous gonadotropin surge were generated by deep sequencing (n=6 small follicle oocyte, 6 small follicle cumulus, 6 large follicle oocyte, 6 large follicle cumulus, 4 spontaneous follcile oocyte, and 5 spontaneous follicle cumulus pools), using Illumina HiSeq 2000 . The sequence reads that passed quality filters were aligned to the Bos taurus reference genome UMD3.1 using Hisat2mapper. FeatureCounts was used to quantify transcript abundance in each library using Bos taurus gene annotation from Ensembl. Differences in gene transcript expression were then analyzed amoung oocyte or cumulus cell follcile classifications using the packages edgeR and DESeq2 in R software. Results: We mapped an average of 30 million sequence reads per sample to the Bos taurus reference genome UMD3.1 and identified 69, 94, and 83 differentially expressed gene transcripts (DEG) among oocyte libraries from small versus large, small versus spontaneous, and large versus spontaneous follicle classifications, respectively. We also identified 128, 98, and 80 DEG among cumulus cell libraries from small versus large, small versus spontaneous, and large versus spontaneous follicles, respectively. Conclusions: Our study represents detailed analysis of RNA-sequencing data generated from in vivo produced cumulus and oocyte samples collected from preovualtory follicles of varied physiological status after exposure to an endogenous or exogenously stimulated gonadotropin surge. The results of our study highlight key differences in the transcriptome of samples from small, large, or spontaneous follicles and provide insight into the reduced embryo quality observed when small follicles undergo a GnRH induced gonadotropin surge.

Dataset Information

Understanding bovine germinal vesicle (GV) oocyte and cumulus cell communication... A systems biology approach

Publications

Differential detergent fractionation for non-electrophoretic eukaryote cell proteomics.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure