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Ovarian Tissue Culture to Visualize Phenomena in Mouse Ovary
Published on: June 19, 2018
Alterations in ovarian function of mice with reduced amounts of KIT receptor
K Reynaud1, R Cortvrindt, J Smitz
1INRA-URA CNRS 1291, PRMD, 37380 Nouzilly, France; and Centre for Reproductive Medicine, Vrije Universiteit Brussel, Laarbeeklaan 101, B-1090 Brussels, Belgium. karine.reynaud@az.vub.ac.be
Abstract:
The KIT receptor, present on oocyte and theca cells in ovarian follicles, and its ligand, KIT LIGAND, produced by granulosa cells, are encoded at the Kit gene and the Mgf gene, respectively. Both Kit and Mgf mutations affect oogenesis and folliculogenesis. In this study, the ovarian function of heterozygous mice with a mutation Kit(W-lacZ) was examined. Firstly, the amounts of KIT and KIT LIGAND proteins in the ovaries of mice at different ages were determined. Secondly, in vivo and in vitro folliculogenesis of wild type and heterozygous mice were compared. Western blotting showed that the amounts of both KIT and KIT LIGAND proteins were decreased in mutant mice. Ovarian follicle populations were counted and more type 5a follicles and fewer type 5b (preantral follicles) were present in ovaries from Kit(W-lacZ/+) ovaries. Furthermore, the relationships between oocyte size and follicle size differed between wild type and heterozygous mice. This finding may be a consequence of altered proliferation of granulosa cells or of altered oocyte growth in mutant mice. Other features of folliculogenesis, such as initiation of follicular growth, total follicle population and follicular atresia, were not affected by the mutation. Analysis of in vitro folliculogenesis did not reveal other differences between wild type and mutant mice. It is concluded that the Kit(W-lacZ) mutation affects the expression of KIT and KIT LIGAND proteins, resulting in alterations in granulosa cell proliferation and/or oocyte growth in preantral follicles.
Insights
The Kit(W-lacZ) mutation in mice reduces KIT and KIT LIGAND proteins, impacting ovarian follicle development. This leads to altered granulosa cell proliferation and oocyte growth in preantral follicles.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Cell Signaling
Background:
- The KIT receptor and its ligand, KIT LIGAND, are crucial for ovarian follicle development.
- Mutations in the Kit and Mgf genes can disrupt oogenesis and folliculogenesis.
Purpose of the Study:
- To investigate the effects of the Kit(W-lacZ) mutation on ovarian function and folliculogenesis in heterozygous mice.
- To analyze changes in KIT and KIT LIGAND protein levels and their impact on follicle development.
Main Methods:
- Western blotting to quantify KIT and KIT LIGAND protein levels in mouse ovaries.
- In vivo and in vitro analysis of folliculogenesis in wild type and Kit(W-lacZ/+) mice.
- Ovarian follicle population counting and characterization of follicle types.
Main Results:
- Kit(W-lacZ/+) mice exhibited decreased KIT and KIT LIGAND protein expression in ovaries.
- A shift in follicle populations was observed, with more type 5a and fewer type 5b (preantral) follicles.
- Differences in the relationship between oocyte size and follicle size were noted in mutant mice.
Conclusions:
- The Kit(W-lacZ) mutation alters KIT and KIT LIGAND protein expression, affecting ovarian function.
- The mutation impacts granulosa cell proliferation and/or oocyte growth during preantral follicle development.
- While some aspects of folliculogenesis were unaffected, the mutation specifically influences preantral follicle dynamics.
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