Related Experiment Video
Updated: Jun 5, 2026
Loss- and Gain-of-function Approach to Investigate Early Cell Fate Determinants in Preimplantation Mouse Embryos
Published on: June 6, 2016
[Study of effect and mechanism of c-myb on the fertilization in mouse]
Ling-feng Wu1, Yue-hui Zheng, Jin Zhang
1Department of Physiology of Jiangxi Medical College, Nanchang 330006, China.
Aim:
To investigate the distribution of c-myb, an oncoprotein, in mouse oocytes-cumulus cell complex and sperm immunohistochemically.
Methods:
To study the effect of c-myb on mouse fertilization in vitro, various concentration of c-myb antisense-oligodeoxynucleotides (c-myb ASODNs) were incubated with sperms and oocytes during fertilization. To explore the possible mechanism involved in fertilization, the relationship between c-myb ASODNs and GABA or dbcAMP or Verapamil or Progesterone in fertilization was also observed by immunohistochemical methods.
Results:
c-myb oncoprotein was observed on the nucleus of cumulus cell and head of sperm. c-myb ASODNs inhibited the rate of fertilization in vitro in a dose-dependent way. The fertilization rates of the control group, low (5 micromol/L), medium (10 micromol/L), high (20 micromol/L) concentration c-myb ASODNs groups and nonsense tat oligodeoxynucleotides (20 micromol/L) group were 34.97%, 30.89%, 20.14%, 16.68%, 34.47%, respectively. All of GABA, Progesterone and dbcAMP inversed the c-myb ASODNs inhibition effects on fertilization rate, but neither of them showed significant effect on the percentages of immunohistochemical stain of Myb on sperm and cumulus cells. By contrast, Verapamil inhibited the fertilization rate. Co-treated with c-myb ASODNs, Verapamil showed synergic inhibiting effects on the fertilization with c-myb ASODNs. Verapamil also inhibited the expression of Myb on head of sperm. The fertilization rates of the control group, medium (10 micromol/L) concentration c-myb ASODNs group, GABA group, P4 group, Verapamil group, dbcAMP group were 34.81%, 22.96%, 40.83%, 39.12%, 7.46%, 40.61%, respectively.
Conclusion:
c-myb ASODNs is closely correlated with fertilization. Verapamil can inhibit fertilization in vitro through regulating Myb expression of sperm, while GABA, dbcAMP and Verapamil may affect the process of fertilization through the way other than Myb expression.
Insights
The oncoprotein c-myb is present in mouse sperm and oocytes. Inhibiting c-myb with c-myb antisense-oligodeoxynucleotides (ASODNs) reduced fertilization rates, with Verapamil showing a synergistic inhibitory effect by regulating sperm Myb expression.
Area of Science:
- Reproductive Biology
- Molecular Oncology
- Cell Biology
Context:
- The c-myb oncoprotein plays a role in cellular processes.
- Understanding its function in gametes is crucial for reproductive research.
- Investigating c-myb's involvement in fertilization provides insights into reproductive mechanisms.
Purpose:
- To determine the distribution of the c-myb oncoprotein in mouse oocytes and sperm.
- To evaluate the impact of c-myb inhibition on in vitro fertilization (IVF).
- To elucidate the mechanisms by which c-myb affects fertilization, including interactions with other compounds.
Summary:
- Immunohistochemical analysis revealed c-myb oncoprotein in mouse cumulus cells and sperm heads.
- Administration of c-myb antisense-oligodeoxynucleotides (ASODNs) dose-dependently inhibited IVF rates.
- GABA, progesterone, and dbcAMP reversed ASODNs' inhibitory effects, suggesting alternative pathways, while Verapamil synergistically inhibited fertilization by downregulating sperm Myb expression.
Impact:
- This study highlights c-myb's critical role in mammalian fertilization.
- Findings suggest c-myb ASODNs as potential modulators of fertilization.
- Verapamil's mechanism of action provides a new avenue for fertility research.
