Verification of the efficacy of deciduae for determining the genetic type of mouse embryos

Author:

Imai Hiroyuki1,Iwamori Naoki2,Iwamori Tokuko23,Matsuya Sumito4,Kano Kiyoshi4,Kusakabe Ken Takeshi1

Affiliation:

1. Laboratory of Veterinary Anatomy, Joint Faculty of Veterinary Medicine Yamaguchi University Yamaguchi Japan

2. Laboratory of Zoology, Graduate School of Agriculture Kyushu University Fukuoka Japan

3. Research Fellow of Japan Society for the Promotion of Science Tokyo Japan

4. Laboratory of Veterinary Embryology, Joint Faculty of Veterinary Medicine Yamaguchi University Yamaguchi Japan

Abstract

AbstractMouse embryos in the early‐implantation stage require manipulation under a microscope. While the extraction of DNA, RNA and proteins from a single sample allows for both determination of genetic type and analysis of gene expression, whole mount analysis is not possible. In this study, we explored the applicability of PCR using extraembryonic tissues, especially the decidual side tissue after isolating the embryos from implantation sites to establish a method for determining the genetic type of embryos. The implantation site was resected at each day from the date of vaginal plug confirmation, separated into embryos and deciduae. Genomic DNA were isolated separately from the embryos and the deciduae. PCR was performed using these genomic DNA, and the band patterns were compared after electrophoresis. As a result, we demonstrated that detecting embryo‐derived cells in the decidua allows determination of the sex and presence of transgenes without harming the mouse embryos themselves, from 8.5 days of age. This method enables the determination of the genetic type of mouse embryos without damaging. This technique would expand the adaptations for analysis of mouse implanted embryos.

Funder

Japan Society for the Promotion of Science

Publisher

Wiley

Subject

General Veterinary,General Medicine

Reference13 articles.

1. Whole‐Mount In Situ Hybridization in Post‐Implantation Staged Mouse Embryos

2. Tracing Gene Expression Through Detection of β-galactosidase Activity in Whole Mouse Embryos

3. A reagent for the single‐step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples;Chomczynski P.;BioTechniques,1993

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