Abstract
Sea urchins are members of a limited group of animals in which meiotic maturation of oocytes is completed prior to fertilization. This is different from oocytes of most animals such as mammals and amphibians in which fertilization reactivates an arrested meiotic cycle. Using a recently developed technique for in vitro maturation of sea urchin oocytes, we analyzed the role of cyclin B, the regulatory component of maturation-promoting factor, in the control of sea urchin oocyte meiotic induction and progression. Oocytes of the sea urchin Lytechinus variegatus accumulate significant amounts of cyclin B mRNA and protein during oogenesis. We analyzed cyclin B synthetic requirements in oocytes and early embryos by inhibiting cyclin B synthesis with DNA and morpholino antisense oligonucleotides. Cyclin B synthesis is not necessary for the entry of G2-arrested oocytes into meiosis; however, it is required for the proper progression through meiotic divisions. Surprisingly, mature sea urchin eggs contain significant cyclin B protein following meiosis that serves as a maternal store for early cleavage divisions. We also find that cyclin A can functionally substitute for cyclin B in early embryos but not in oocytes. These studies provide a foundation for understanding the mechanism of meiotic maturation independent of the zygotic cell cycle.
| Original language | English |
|---|---|
| Pages (from-to) | 258-275 |
| Number of pages | 18 |
| Journal | Developmental Biology |
| Volume | 256 |
| Issue number | 2 |
| DOIs | |
| State | Published - Apr 15 2003 |
Funding
We thank past and present members of PRovidence Institute of Molecular Oogenesis for unwavering support and assistance, especially Emma R. Green for help in cloning sea urchin GAPDH, Dr. Jacqueline M. Brooks for aid in developing the RT-PCR technique, and Audrey S. Howell for contribution of an EST clone spanning cyclin B 3′ UTR. Our gratitude extends to Dr. Bradley J. Schnackenberg for supplying protocols, reagents, and assistance in performing histone H1 kinase assays, as well as to Dr. Bettina Meier, Julie A. Boerckel, and Özlem Yildiz for help in performing the activity assays. We appreciate special assistance of Wayne Baumgartner of CCAT. This research was supported by NIH and NSF grants to G.M.W.
Keywords
- Cyclin B
- Egg
- MPF, Embryo
- Meiosis
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