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Injecting Gryllus bimaculatus Eggs
Published on: August 22, 2019
Injection of an antibody against a p21 c-Ha-ras protein inhibits cleavage in axolotl eggs
E Baltus1, J Hanocq-Quertier, F Hanocq
1Département de Biologie Moléculaire, Université Libre de Bruxelles, Rhode-St.-Genèse, Belgium.
Abstract:
The presence of a ras protein was demonstrated in cleaving axolotl eggs by selective immunoprecipitation with a polyclonal antibody against a peptide encoded by the c-Ha-ras oncogene, cellular homolog of the v-Ha-ras oncogene of Harvey rat sarcoma virus. Injection of this antibody into axolotl oocytes subjected to progesterone treatment does not prevent meiotic maturation. Injection of the same antibody into a blastomere of axolotl eggs at the 2- or 4-cell stage causes cleavage arrest in the descendants of the injected blastomere. Cytological observations of the injected eggs show, in the arrested blastomeres, enlarged nuclei always surrounded by an intact nuclear envelope and containing uncondensed chromatin. The possible role of ras protein in meiosis and mitosis is discussed.
Insights
Ras protein is present in axolotl eggs and is crucial for cell division. Inhibiting ras protein stops early embryonic cell cleavage but does not affect meiotic maturation.
Area of Science:
- Developmental Biology
- Molecular Biology
- Oncogene Research
Background:
- Ras proteins are key signaling molecules involved in cell growth and differentiation.
- The c-Ha-ras oncogene is a cellular homolog of the viral v-Ha-ras oncogene.
- Understanding the role of ras proteins in early development is crucial for developmental biology.
Purpose of the Study:
- To investigate the presence and function of ras protein in early axolotl development.
- To determine the role of ras protein in meiotic maturation and early embryonic cleavage.
- To explore the potential involvement of ras protein in cell cycle regulation.
Main Methods:
- Selective immunoprecipitation using a polyclonal antibody against a peptide encoded by the c-Ha-ras oncogene.
- Injection of the anti-ras antibody into axolotl oocytes and early-stage blastomeres.
- Cytological observation of injected cells to assess meiotic maturation and cleavage progression.
Main Results:
- Ras protein was detected in cleaving axolotl eggs.
- Antibody injection into oocytes did not inhibit progesterone-induced meiotic maturation.
- Antibody injection into 2- or 4-cell stage blastomeres resulted in cleavage arrest in descendant cells.
- Arrested blastomeres exhibited enlarged nuclei with uncondensed chromatin and intact nuclear envelopes.
Conclusions:
- Ras protein plays a critical role in regulating mitosis during early axolotl embryogenesis.
- The function of ras protein in mitosis appears distinct from its role in meiotic maturation.
- Ras protein's involvement in cell cycle progression warrants further investigation in both normal development and oncogenesis.
