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  • Effects of WT1 down-regulation on oocyte maturation and preimplantation embryo development in pigs.

Effects of WT1 down-regulation on oocyte maturation and preimplantation embryo development in pigs.

Reproduction (Cambridge, England) (2014-07-18)
Fei Gao, Jiyu Guan, Limei Liu, Sheng Zhang, Peipei An, Anran Fan, Guangqi Song, Peng Zhang, Tianchuang Zhao, Bo Tang, Xueming Zhang, Ziyi Li
ABSTRACT

The Wilms' tumour 1 (WT1) gene originally identified as a tumour suppressor associated with WTs encodes a zinc finger-containing transcription factor that is expressed in multiple tissues and is an important regulator of cellular and organ growth, proliferation, development, migration and survival. However, there is a deficiency of data regarding the expression and function of WT1 during oocyte maturation and preimplantation embryonic development. Herein, we sought to define the expression characteristics and functions of WT1 during oocyte maturation and preimplantation embryonic development in pigs. We show that WT1 is expressed in porcine oocytes and at all preimplantation stages in embryos generated by ICSI. We then evaluated the effects of down-regulating WT1 expression at germinal vesicle and early ICSI stages using a recombinant plasmid (pGLV3-WT1-shRNA). Down-regulation of WT1 did not affect oocyte maturation but significantly decreased preimplantation embryonic development and increased apoptosis in blastocysts. These results indicate that WT1 plays important roles in the development of porcine preimplantation embryos.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
MISSION® esiRNA, targeting human WT1
Roche
In Situ Cell Death Detection Kit, POD, sufficient for ≤50 tests
Sigma-Aldrich
Goat Anti-Rabbit IgG (H+L) Antibody, FITC conjugate, Upstate®, from goat
Sigma-Aldrich
Goat Anti-Rabbit IgG Antibody, (H+L) Rhodamine conjugate, 1.0 mg/mL, Chemicon®