title: The double-edged sword of the mammalian oocyte--advantages, drawbacks and approaches for basic and clinical analysis at the single cell level.
abstract: Oocytes are usually the largest cells in the body and as such offer unique opportunities for single-cell analysis. Unfortunately, these cells are also some of the rarest in the mammalian female, usually necessitating single-cell analysis. In cases of infertility in humans, determining the quality of the oocyte is often restricted to a morphological analysis or to the study of cellular behaviors in the developing embryo. Minimally invasive approaches could greatly assist the clinician to prioritize oocytes for fertilization or following fertilization, which embryo to transfer back into the woman. Transcriptomics of human and mouse oocytes may have great utility, and recently it was learned that the polar body faithfully reflects the transcript prevalence in the oocyte. The polar body may thus serve as a minimally invasive proxy for an oocyte in the clinic. In the mouse, the transcriptomes of oocytes from mice of the same strain are markedly similar; no significant differences are apparent in transcript prevalence or identity. In human oocytes however, the transcript pool is highly variable. This is likely the result of different histories of each oocyte, in the age of the donor woman, the different hormonal exposures and the prolonged time from specification of the primary oocyte to the fully grown and ovulated egg. This variability in human oocytes also emphasizes the need for cell-by-cell analysis of the oocytes in vitro; which oocytes have a better potential for fertilization and development? To this end, new imaging capabilities are being employed. For example, a single-cell analytical device for oocytes (the simple perfusion apparatus, or SPA) enables investigators to load multiple oocytes in individual wells, to visualize them on the microscope and to use controlled temperature and media flow by perfusion for optimal clinical applications. Recently, developed Raman microspectroscopy approaches suggest that this imaging modality may enable more in-depth analysis of the molecular characteristics of an oocyte that, in combination with the SPA and transcriptomic approaches, might assist the clinician to prioritize more effectively human oocytes and embryos for transfer into women. This review is intended to update the reader on the status of the examination of single oocytes from a variety of approaches and to emphasize areas that may be primed for advancement in the near future.
authors.affiliation: Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, Women & Infants Hospital, Warren Alpert Medical School, Brown University, 101 Dudley, Fl1, Providence, RI 020905, USA.
authors.forename: L M
authors.initials: LM
authors.lastname: Brayboy
authors.affiliation: Department of Molecular and Cellular Biology & Biochemistry, Brown University, 185 Meeting Street, Providence, RI 02912, USA rhet@brown.edu.
authors.forename: G M
authors.initials: GM
authors.lastname: Wessel
mesh_terms: D000818:Animals; D048429:Cell Size; D047108:Embryonic Development; D005260:Female; D023281:Genomics; D006801:Humans; D055442:Metabolome; D009865:Oocytes / Q000166:cytology*; D020543:Proteome; D059010:Single-Cell Analysis; D014015:Tissue Banks; D059467:Transcriptome
chemical_list: D020543:Proteome
keywords: Raman microspectroscopy; oocyte; oocyte gene expression; simple perfusion apparatus; single-cell analysis; transcriptome