The NOBOX protein becomes undetectable in developmentally competent antral and ovulated oocytes.

The oocyte-specific NOBOX protein is an important player during oocyte growth. Its absence in Nobox-/- mice arrests the transition from primordial to growing follicles and down-regulates the expression of a number of genes, including Oct4, a transcription factor crucial in the acquisition of oocyte developmental competence. Despite its role during folliculogenesis, a clear description of the expression of NOBOX throughout oocyte growth is lacking. Here, we have analysed the pattern of expression of both the Nobox gene (qRT-PCR) and its protein (immunofluorescence) during folliculogenesis, classifying the oocytes based on their size (six classes: 10-30, 31-40, 41-50, 51-60, 61-70, 71-80 mm) and chromatin organisation (NSN, Non Surrounded Nucleolus or SN, Surrounded Nucleolus). Significant differences were observed in Nobox transcription in the group of 41-50 mm (NSN > SN), 71-80 mm (NSN > SN) and in developmentally incompetent metaphase II-derived NSN (MIINSN) or competent metaphase II-derived SN (MIISN) oocytes (MIINSN > MIISN). The NOBOX protein is expressed throughout oocyte growth in the nucleus of ovarian NSN and in MIINSN oocytes; in contrast, beginning with SN oocytes of 61-70 mm, it becomes almost undetectable. Our data, while being in line with the hypothesis of a regulative role of NOBOX on Oct4 gene expression at the primordial/primary stage, when both transcription factors are coincidentally expressed, also indicate that this role might not be maintained in the subsequent growing stages. Furthermore, the sharp difference of NOBOX expression in developmentally incompetent or competent oocytes makes this protein a putative marker of their quality.