Skip to main content

Oocytes Microinjection Assay to Study the MAP-Kinase Cascade

  • Protocol
  • 553 Accesses

Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 84))

Abstract

The oocytes of several organisms—most frequently those of the African clawed toad Xenopus laevis—have been used for many years as an excellent system to study regulation of transcription, translation, protein modification processes, secretion, and protein compartmentalization, as well as the expression of heterologous-membrane receptors and their association to specific signaling cascades. Full-grown oocytes are large cells (over 1.2 mm in diameter) that are arrested in late-G2 phase of the first meiosis (Meiosis I), and must progress after physiologrcal stimulus by progesterone to the second meiotrc metaphase (Meiosis II) before fertilization takes place. This process, called oocyte maturation or germinal vesicle breakdown (GVBD), can be easily visualized by the appearance of a small white spot in the animal pole, a consequence of the dissociation of the nuclear envelope. After GVBD is completed, if the oocytes have been fertilized, DNA synthesis takes places with the consequent initiation of the Meiosis II.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Maller J. L (1990) Xenopus oocytes and the biochemistry of cell dlvlslon Biochemistry 29, 3157–3166

    Article  PubMed  CAS  Google Scholar 

  2. Norbury C and Nurse P (1992) Animal cell cycles and their control Annu Rev Biochem 61, 441–470

    Article  PubMed  CAS  Google Scholar 

  3. Jacobs T (1992) Control of the cell cycle Devl Biol 153, 1–15

    Article  CAS  Google Scholar 

  4. Lacal J C. and Carnero A (1994) Regulation of Ras proteins and then mvolvement in signal transduction pathways Oncology Reports 1, 611493

    Google Scholar 

  5. Bauheu E. E, Godeau F, Schorderet M., and Schorderet-Slatkme S (1978) Steroid-mduced melotic dlvlslon in Xenopus laevls oocytes surface and calcium Nature, 275, 593–598

    Article  Google Scholar 

  6. Maller J L and Koontz J. W (1981) A study of the mduction of cell divrsion inamphibian oocytes by insulin. Devl Biol 85, 309–316

    Article  CAS  Google Scholar 

  7. Masui Y. and Markert C L (1971) Cytoplasmic control of nuclear behavior during meiotic maturation of frog oocytes J Exp Zoo1 177, 129–146

    Article  CAS  Google Scholar 

  8. Smith L D (1989) The mduction of oocyte maturation. transmembrane signaling events and regulation of the cell cycle. Development 107, 685–699.

    PubMed  CAS  Google Scholar 

  9. Sagata N., Daar I, Oskarsson M., Schowalter S. D, and Vande-Woude G. F (1989) The product of the mos proto-oncogene as a candidate “mitiator” for oocyte maturation Science 245, 643–645

    Article  PubMed  CAS  Google Scholar 

  10. Btrchmeyer C, Broek D, and Wigler M. (1985) Ras proteins can induce meiosis in Xenopus oocytes Cell. 43, 615–620

    Google Scholar 

  11. Hollinger T. G. and Alvarez I M. (1982) Trifluoperazme-induced meiotic maturation in Xenopus laevis J Exp Zoo1 224, 461–464

    Article  CAS  Google Scholar 

  12. Carnero A. and Lacal J C (1993) Phospholipase-induced maturation of Xenopus laevrs oocytes. Mitogemc activity of generated metabolites. J Cell Biochem 52, 440–448

    Article  PubMed  CAS  Google Scholar 

  13. Cicirelli M. F and Smith L D. (1987) Do calcmm and calmodulin trigger maturation in amphibian oocytes? Devl Biol 121, 48–57

    Article  CAS  Google Scholar 

  14. Brizuela L, Draetta G, and Beach D. (1987) p13sucl acts in the fission yeast cell division cycle as a component of the p34cdc2 protein kinase EMBO J 6, 3507–3514.

    PubMed  CAS  Google Scholar 

  15. Kameshita I. and Fujisawa H. (1989) A sensitive method for detection of calmodulin-dependent protein kinase II activity in sodmm dodecyl sulfate polyacrylamide gel. Anal Biochem 183, 139–143

    Article  PubMed  CAS  Google Scholar 

  16. Lacal J. C (1990) Diacylglycerol production in Xenopus laevls oocytes after mmromJection of p2lras proteins is a consequence of activation of phosphatidylcholine metabolism. Mol Cell Biol 10, 333–340.

    PubMed  CAS  Google Scholar 

  17. Carnero A., Liyanage M., Stabel S., and Lacal J. C. (1995) Evidence for different signalling pathways of PKC and ras-p21 m Xenopus oocytes. Oncogene 11, 1541–1547

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc.

About this protocol

Cite this protocol

Lacal, J.C. (1998). Oocytes Microinjection Assay to Study the MAP-Kinase Cascade. In: Bar-Sagi, D. (eds) Transmembrane Signaling Protocols. Methods In Molecular Biology™, vol 84. Humana Press. https://doi.org/10.1385/0-89603-488-7:139

Download citation

  • DOI: https://doi.org/10.1385/0-89603-488-7:139

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-488-4

  • Online ISBN: 978-1-59259-568-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics