Discovery of putative oocyte quality markers by comparative ExacTag proteomics

Abstract

Purpose Identification of the biomarkers of oocyte quality, and developmental and reprogramming potential is of importance to assisted reproductive technology in humans and animals. Experimental design PerkinElmer ExacTag™ Kit was used to label differentially proteins in pig oocyte extracts (oocyte proteome) and pig oocyte-conditioned in vitro maturation media (oocyte secretome) obtained with high- and low-quality oocytes. Results We identified 16 major proteins in the oocyte proteome that were expressed differentially in high- versus low-quality oocytes. More abundant proteins in the high-quality oocyte proteome included kelch-like ECH-associated protein 1 (an adaptor for ubiquitin-ligase CUL3), nuclear export factor CRM1 and ataxia-telangiectasia mutated protein kinase. Dystrophin (DMD) was more abundant in low-quality oocytes. In the secretome, we identified 110 proteins, including DMD and cystic fibrosis transmembrane conductance regulator, two proteins implicated in muscular dystrophy and cystic fibrosis, respectively. Monoubiquitin was identified in the low-quality-oocyte secretome. Conclusions and clinical implications A direct, quantitative proteomic analysis of small oocyte protein samples can identify potential markers of oocyte quality without the need for a large amount of total protein. This approach will be applied to discovery of non-invasive biomarkers of oocyte quality in assisted human reproduction and in large animal embryo transfer programs.

From global proteome profiling to single targeted molecules of follicular fluid and oocyte: contribution to embryo development and IVF outcome

“…The more abundant proteins in high oocyte quality proteome were mainly Kelchlike ECH-associated protein 1 (an adaptor for ubiquitin-ligase CUL3), nuclear export factor CRM1 and Ataxia-telangiectasiamutated protein kinase. In the opposite, Dystrophin (DMD) was more abundant in low-quality oocytes [113].…”

“…Stabilization of the COC matrix and oocyte quality Bovine [101] Oocyte Ubiquitin ligase, nuclear export factors protein, kinase protein Oocyte quality Mouse [113] LAMP1,2, Glyoxylase 1 (GLO1) and ubiquitin Oocyte quality, maturation and competence Bovine and mouse [119,167] TCTP Overexpression increases oocyte maturation Bovine [136] CD55, CD9, CD81, GRP94, BiP (GRP78), ORP150 (hypoxia up-regulated 1), calreticulin, calnexin, PDI Important for fertilization Mouse [119,168,169] HSP70, (HSP1b), HSP90a Important for fertilization Mouse [169][170][171] avb3 integrin, a6b1 integrin Important for fertilization Bovine [169,171] Kelch-like ECH-associated protein 1 (KEAP1) Oocyte quality and maturation Mouse [169,172] Dystrophin (DMD) Over-expression decreases oocyte quality Mouse [113] Bisphosphoglyceratemutase (BPGM) Oocyte maturation and considered as maternal housekeeping protein Mouse [169,172] technology. As a result of assessing murine MII oocytes, beginning with 156-380 unique proteins using 2D electrophoresis coupled to MS [73,74] then 625 proteins with 1D-SDS-PAGE and RP-LC-MS/MS [19] until reaching a profile of 3699 proteins [75], the proteomic obtained information is associated with early development [75].…”

“…A defined dose of gonadotrophins in in vitro maturation (IVM) medium revealed significant differences in expression for 10 proteins [111]. Powel et al [113] reported that eight of 110 identified proteins were differentially abundant between low-and-high quality oocytes depending on gonadotrophins doses in IVM culture medium. The more abundant proteins in high oocyte quality proteome were mainly Kelchlike ECH-associated protein 1 (an adaptor for ubiquitin-ligase CUL3), nuclear export factor CRM1 and Ataxia-telangiectasiamutated protein kinase.…”

The Oocyte Determinants of Early Reprogramming

“…It is important to note that DJ-1 has also been identified as important factor for reprogramming by Miyamoto and colleagues (Miyamoto et al 2011). Furthermore, quality markers, such as kelch-like Ech-associated protein 1 and CRM1, a nuclear export factor, were found to be highly abundant in high-quality porcine oocytes (Powell et al 2010). Another study on porcine oocytes showed that oocytes with the first polar body collected at 42 h of in vitro maturation were more supportive of cloned embryo development than oocytes with the first polar body collected at 33 h (Kong et al 2014).…”

Pig proteomics: A review of a species in the crossroad between biomedical and food sciences

“…Such proteins include structural proteins such as actin or tubulin, as well as initiation factors, elongation factor 1, endoplasmic reticulum luminal protein or spermidine aminopropyltransferase. More recently, Powell and colleagues [14], using ExacTag™ labeling kit proteomics, have identified biomarkers of oocyte quality and developmental and reprogramming potential by comparing high and low quality oocytes. The study was conducted at the level of the oocyte itself, as well as the secretome.…”