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Activation of pig and cattle oocytes by butyrolactone I: morphological and biochemical study.
- Source :
-
Zygote (Cambridge, England) [Zygote] 2002 Feb; Vol. 10 (1), pp. 47-57. - Publication Year :
- 2002
-
Abstract
- In this study a specific inhibitor of cyclin-dependent kinases (cdks), butyrolactone I (BL I), was used for activation of pig and cattle metaphase II (MII) oocytes. BL I at a concentration of 100 microM was able to induce activation of both pig and cattle MII oocytes in a manner dependent on exposure time; however, precise timing of BL I exposure was required for the best activation results. The optimum activation rates were obtained when cattle MII oocytes were treated for 5 h with BL I and subsequently for 3-11 h in control medium, and pig MII oocytes for 8 h in BL I and then for 8-16 h in control medium; the percentage of activated oocytes after such treatment varied between 55% and 74% and between 53% and 81% for cattle and pig oocytes, respectively. Shorter exposures to BL I led to re-entry of the oocytes to the metaphase state in 35-50% of oocytes, the remaining oocytes forming a pronuclear stage; longer exposure to BL I led to increased numbers of oocytes being abnormal or degenerated. The behaviour of histone H1 kinase and mitogen activated protein (MAP) kinase, also measured during the experiment, reflected the morphological changes in the oocytes: both were inactivated after BL I treatment, though the inactivation of histone H1 kinase occurred 2 h ahead of that of MAP kinase. However, in the oocytes treated for a shorter time with BL I, with the reoccurrence of condensed chromatin in proportion of the oocytes cultured in control medium after BL I treatment, both kinases became reactivated. Taken together, these results suggest the possibility of using BL I for activation and cloning experiments in both species.
- Subjects :
- Animals
Blotting, Western
CDC2 Protein Kinase antagonists & inhibitors
CDC2 Protein Kinase metabolism
Cattle
Cells, Cultured
Cyclin-Dependent Kinases antagonists & inhibitors
Cyclin-Dependent Kinases metabolism
Histones metabolism
Mitogen-Activated Protein Kinases antagonists & inhibitors
Mitogen-Activated Protein Kinases metabolism
Myelin Basic Protein antagonists & inhibitors
Myelin Basic Protein metabolism
Oocytes cytology
Swine
4-Butyrolactone analogs & derivatives
4-Butyrolactone pharmacology
Oocytes drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 0967-1994
- Volume :
- 10
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Zygote (Cambridge, England)
- Publication Type :
- Academic Journal
- Accession number :
- 11964091
- Full Text :
- https://doi.org/10.1017/s0967199402002071