Synchronization of In Vitro Maturation in Porcine Oocytes

Methods Mol Biol. 2017:1524:255-264. doi: 10.1007/978-1-4939-6603-5_16.

Abstract

When removed from the follicles, during the 44 h process of in vitro maturation (IVM) fully grown porcine oocytes resume meiosis spontaneously from the late diplotene stage of the first meiotic prophase and proceed to the metaphase-II (MII) stage at which they remain arrested until fertilization. However, the spontaneous resumption may start at various times causing heterogeneity in the nuclear stage and also in cytoplasmic characteristics within a population. Those oocytes that reach the MII stage earlier than others undergo an aging process which is detrimental for further embryo development. The synchronization of nuclear progression of porcine oocytes can be achieved by a transient inhibition of meiotic resumption during the first 20-22 h of IVM by the elevation of intracellular levels of cyclic adenosine monophosphate (cAMP) using the cellular membrane-permeable analog of cAMP, dibutyryl cyclic AMP. A simple and efficient protocol for such treatment is described below.

Keywords: In vitro maturation; Oocyte; Porcine; Synchronization; cAMP.

MeSH terms

  • Animals
  • Bucladesine / metabolism
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Cyclic AMP / metabolism
  • Meiosis / genetics
  • Meiosis / physiology
  • Metaphase / genetics
  • Metaphase / physiology
  • Oocytes / cytology*
  • Oocytes / physiology
  • Swine

Substances

  • Bucladesine
  • Cyclic AMP