Gene expression in mouse ovarian follicle development in vivo versus an ex vivo alginate culture system

Reproduction. 2011 Aug;142(2):309-18. doi: 10.1530/REP-10-0481. Epub 2011 May 24.

Abstract

Ovarian follicle maturation results from a complex interplay of endocrine, paracrine, and direct cell-cell interactions. This study compared the dynamic expression of key developmental genes during folliculogenesis in vivo and during in vitro culture in a 3D alginate hydrogel system. Candidate gene expression profiles were measured within mouse two-layered secondary follicles, multi-layered secondary follicles, and cumulus-oocyte complexes (COCs). The expression of 20 genes involved in endocrine communication, growth signaling, and oocyte development was investigated by real-time PCR. Gene product levels were compared between i) follicles of similar stage and ii) COCs derived either in vivo or by in vitro culture. For follicles cultured for 4 days, the expression pattern and the expression level of 12 genes were the same in vivo and in vitro. Some endocrine (cytochrome P450, family 19, subfamily A, polypeptide 1 (Cyp19a1) and inhibin βA subunit (Inhba)) and growth-related genes (bone morphogenetic protein 15 (Bmp15), kit ligand (Kitl), and transforming growth factor β receptor 2 (Tgfbr2)) were downregulated relative to in vivo follicles. For COCs obtained from cultured follicles, endocrine-related genes (inhibin α-subunit (Inha) and Inhba) had increased expression relative to in vivo counterparts, whereas growth-related genes (Bmp15, growth differentiation factor 9, and kit oncogene (Kit)) and zona pellucida genes were decreased. However, most of the oocyte-specific genes (e.g. factor in the germline α (Figla), jagged 1 (Jag1), and Nlrp5 (Mater)) were expressed in vitro at the same level and with the same pattern as in vivo-derived follicles. These studies establish the similarities and differences between in vivo and in vitro cultured follicles, guiding the creation of environments that maximize follicle development and oocyte quality.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Validation Study

MeSH terms

  • Alginates / chemistry
  • Alginates / metabolism*
  • Animals
  • Cells, Cultured
  • Chemical Phenomena
  • Crosses, Genetic
  • Cumulus Cells / physiology
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental*
  • Glucuronic Acid / chemistry
  • Glucuronic Acid / metabolism
  • Hexuronic Acids / chemistry
  • Hexuronic Acids / metabolism
  • Hydrogel, Polyethylene Glycol Dimethacrylate
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred CBA
  • Oogenesis*
  • Ovarian Follicle / physiology*
  • Ovulation Induction
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Tissue Culture Techniques*

Substances

  • Alginates
  • Hexuronic Acids
  • RNA, Messenger
  • Hydrogel, Polyethylene Glycol Dimethacrylate
  • Glucuronic Acid