Transition-metal-mediated dearomatization reactions.
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Biomedical subjects
A R Pape
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Expression of interleukin-1 system mRNA in single blastomeres from human preimplantation embryos.
Gathering knowledge about the molecular events during preimplantation development is one of the most important challenges in in-vitro fertilization (IVF). The interleukin-1 (IL-1) system has been shown to be intimately involved in embryonic implantation. The aim of our study was to detect the major components of the IL-1 system in single blastomeres from human preimplantation embryos and to relate our findings to the further development of the biopsied embryos in vitro. Single blastomeres were removed from morphologically normal embryos obtained from dipronuclear zygotes and examined by reverse transcription (RT)-nested polymerase chain reaction (PCR). Expression of beta-actin (external standard), IL-1beta, IL-1 receptor antagonist (IL-1ra) and IL-1 receptor (IL-1R) type I mRNA were related to embryonic development and IVF outcome. Blastomeres from 12 embryos were examined: beta-actin and IL-1R type I mRNA were detected in all blastomeres (100%) whereas IL-1beta could be detected in only nine of the blastomeres (75%). IL-1ra was expressed in only two (17%) of the blastomeres and those were simultaneously positive for IL-1beta. Both IL-1ra positive embryos were arrested in development before reaching blastocyst stage. Five embryos (three of them IL-1beta mRNA positive and two IL-1beta mRNA negative) were transferred as blastocysts; none of the transfers resulted in a pregnancy. We postulate that embryos expressing IL-1ra mRNA in a detectable amount appear more likely to be arrested in early developmental stages.
Base Sequence↗
Single blastomeres within human preimplantation embryos express different amounts of messenger ribonucleic acid for beta-actin and interleukin-1 receptor type I.
Gaining knowledge about the physiological timetable of gene expression during preimplantation embryo development is crucial, and a better understanding of cytokine and growth factor expression in early embryonic development could lead to improved in vitro culture conditions and enhance in vitro fertilization implantation rates. Our aim was to detect the patterns and levels of two messenger ribonucleic acids [mRNAs; beta-actin and interleukin-1 receptor type I (IL-1R tI)] in single human blastomeres by RT-nested PCR and to compare possible variations in the gene expression both between different embryos and in multiple blastomeres within the same embryo. Single blastomeres from nine human tripronucleic preimplantation embryos were examined by one round of RT and two rounds of nested competitive PCR. Beta-actin mRNA was detected in each blastomere, and IL-1R tI mRNA was found in 72% of the blastomeres examined. Beta-actin was expressed at a level of 511-12185 molecules of complementary DNA/blastomere, and IL-1R tI was expressed at a level of 2-290 molecules of complementary DNA/blastomere. Our results suggest that the mRNA pattern of an embryo cannot be reliably quantitated from the mRNA pattern of a single blastomere and therefore imply limitations for the use of this method for preimplantation diagnosis.
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