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Differences in gene expression patterns between somatic cell nuclear transfer embryos constructed with either rabbit
Fukashi Inoue1, Junichi Matsuda, Katsuhiro Ohkoshi
1Development and Differentiation Laboratory, Developmental Biology Department, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan. fukashi@affrc.go.jp
Animal Reproduction Science
|August 10, 2005
Summary
Somatic cell nuclear transfer (SCNT) offspring success depends on donor cells. In vitro culture conditions altered rabbit granulosa cells, impacting gene expression in SCNT embryos, though developmental potential remained unchanged.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Cell Biology
Background:
- Somatic cell nuclear transfer (SCNT) efficiency is influenced by donor cell characteristics.
- In vitro culture can induce changes in donor cells, potentially affecting SCNT outcomes.
- Rabbit granulosa cells (rGC) are a relevant model for studying donor cell modifications.
Purpose of the Study:
- To investigate how in vitro culture conditions alter gene expression in rabbit granulosa cells (rGC).
- To determine if these culture-induced changes in rGC affect gene expression patterns in subsequent somatic cell nuclear transfer (SCNT) embryos.
- To assess the impact of donor cell modifications on the developmental potential of SCNT embryos.
Main Methods:
- Rabbit granulosa cells (rGC) were cultured with or without hCG, creating two distinct cell types (cGC+ and cGC-).
- These modified cells and fresh rGC were used as donor cells for SCNT.
- Gene expression analysis (quantitative RT-PCR) of specific genes (bmp4, fgfr2, gata4, oct3/4) in SCNT embryos and fertilized controls.
Main Results:
- Cultured cells (cGC+ and cGC-) exhibited altered expression of p450scc, bmp4, fgfr2, and gained collagenIV compared to fresh rGC.
- SCNT oocyte developmental potency and blastocyst cell numbers were not significantly affected by donor cell type.
- Differential expression of bmp4, fgfr2, and gata4 was observed in SCNT embryos depending on the donor cell origin, with oct3/4 universally expressed.
Conclusions:
- In vitro culture conditions induce characteristic changes in rabbit granulosa cells.
- These donor cell modifications significantly influence the gene expression profiles of resulting SCNT embryos.
- While gene expression patterns are altered, the overall developmental competence of SCNT embryos is not compromised by these specific donor cell changes.