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Genes and proteins related to IMP2 pathway in maternal-zygotic transformation and applications thereof

A maternal and protein technology, applied in the field of assisted reproduction of mammals, especially humans, can solve problems that need to be studied and the role is unclear

Active Publication Date: 2020-02-21
SHANDONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the role of Imp2 in mouse embryonic development remains unclear, especially the contribution of maternal Imp2 to mRNA and protein production during embryogenesis remains to be investigated

Method used

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  • Genes and proteins related to IMP2 pathway in maternal-zygotic transformation and applications thereof
  • Genes and proteins related to IMP2 pathway in maternal-zygotic transformation and applications thereof
  • Genes and proteins related to IMP2 pathway in maternal-zygotic transformation and applications thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Patient recruitment and ethics review

[0068] The research for this application was approved by the Review Committee of the Institute of Reproductive Medicine, Shandong University. All methods described in this application were performed in accordance with the approved guidelines and regulations of the Institute of Reproductive Medicine, Shandong University. The in vitro cultured oocytes used came from clinically discarded immature eggs (GV stage) in the Reproductive Hospital Affiliated to Shandong University. A formal informed consent was obtained from each patient before the experiments related to human beings.

Embodiment 2

[0069] Embodiment 2 experimental method and reagent

[0070] Oocyte and Embryo Collection and Microinjection

[0071] 24-28 day old mice were superstimulated for 44 hours with 5 IU pregnant mare serum gonadotropin (PMSG) and 5 IU human chorionic gonadotropin (hCG). Oocytes were harvested and cultured in small drops of M16 medium (M7292; Sigma-Aldrich), overlaid with mineral oil and maintained at 37°C in 5% CO 2 middle. For collection of fertilized eggs and embryos, control and Imp2 - / - Females were mated with adult WT males after hCG injection. For fertilized egg collection, fallopian tubes were punctured, while for embryo collection, the uterus was flushed at indicated time points after hCG administration. For microinjection, mRNA was transcribed in vitro using the mMESSAGE mMACHINE SP6 Transcription Kit (Invitrogen, AM1450). The siRNAs were obtained from RiboBio and the sequences are given in Table 2.

[0072] Fertilized egg culture, embryo transfer and fertility asses...

Embodiment 3

[0107] Example 3 High expression of Imp2 in mouse oocytes and early embryos

[0108] The protein and mRNA profiles of IMP2 in mouse oocytes and early embryos were determined by Western blotting and quantitative real-time PCR (qRT-PCR), respectively. Transcripts for the mRNA-binding protein IMP2 were found to be highly expressed in mouse oocytes and early embryos. Expression is strongest at the germinal vesicle (GV) stage and is significantly lower in MII oocytes. The expression decreased further after fertilization, and completely absent at the blastocyst stage ( figure 1 a).

[0109] Immunofluorescence staining showed that IMP2 was localized in the cytoplasm of oocytes and preimplantation embryos ( figure 1 b). IMP2 expression is uniformly distributed at the oocyte stage but undergoes dynamic changes during zygote development. Morula and blastocyst stages showed IMP2 expression at the outer edge of the blastomere ( figure 1 b), and Western blot analysis further confirme...

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Abstract

The present invention provides a method or a kit or a device for evaluating embryos used for in vitro fertilization-embryo transplantation through protein or gene detection of a maternal IMP2 pathwayand provides the method or the kit or the device for assessing risks of the in vitro fertilization-embryo transplantation, especially pregnancy loss during an embryo-implantation early period by the protein or gene detection of the maternal IMP2 pathway. The method or the kit or the device is particularly suitable for assessing the risk of the pregnancy loss during the embryo-implantation early period caused by abnormal activation of zygote genomes.

Description

technical field [0001] The invention relates to the field of assisted reproduction of mammals, especially humans. Specifically, the present invention relates to a method or a kit or a device for evaluating the embryo quality of IVF-ET or the risk of IVF-ET by detecting the protein or gene of the maternal IMP2 pathway of mammalian embryos. The method or kit or device of the present invention is particularly suitable for assessing the risk of pre-implantation pregnancy loss caused by abnormal activation of the zygotic genome. Background technique [0002] Mammalian eggs are the most important cells in the female body, generally in a quiescent state, but after fertilization, egg cells are reprogrammed into highly specialized totipotent fertilized eggs. This reprogramming process takes the embryo into a developmental process through highly specialized, proliferative states and increasingly differentiated stages, ultimately leading to the development of new individuals. During ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888G01N33/68
CPCC12Q1/6888G01N33/68C12Q2600/156
Inventor 刘洪彬陈子江赵跃然马金龙耿玲路钢刘奎穆罕默德·塔希尔黄涛
Owner SHANDONG UNIV
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