A mouse embryo culture medium which would allow for in vitro development from 1-cell stage to blastocyst stage could offer many benefits for human research. Previous researchfrom our lab has demonstrated a mouse embryo culture medium named CZB seems to allow for in vivo-like conditions for development. Compared to other commonly used mouse embryo culture media, CZB medium promotes a higher frequency of 1 cell mouse embryos developing to blastocyst stage (Chatot et 1989). A key difference between CZB and other mouse embryo culture media is that CZB contains the amino acid glutamine metabolism is glutamate dehydrogenase (GDH). In order to determine if CZB cultured embryos follow in vivo-like patterns of gene expression for GDH, a quantitative competitive RT-PCR system was designed. A mutant GDH mRNA template was created which lacked a specific restriction enzyme site and was used as a competitive template in quantitative RT-PCR. This system was used to determine the amount of GDH mRNA present in in vivo grown blastocyst stage mouse embryos. It was determined that the amount of GDH mRNA present in in vivo blastocyst stage embryos was 282 fg/embryo. It is believed this system will also allow for quantitation of GDH mRNA in the earlier preimplantation stages of in vivo grown embryos, as well as the preimplantation stages 2-cell to blastocyst of CZB cultured embryos. / Department of Biology
| Identifer | oai:union.ndltd.org:BSU/oai:cardinalscholar.bsu.edu:handle/184836 |
| Date | January 1994 |
| Creators | Lawry, John R. |
| Contributors | Ball State University. Dept. of Biology., Chatot, Clare L. |
| Source Sets | Ball State University |
| Detected Language | English |
| Format | vii, 98 leaves : ill. ; 28 cm. |
| Source | Virtual Press |
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