Analysis of G protein alpha subunit mRNA abundance in preimplantation mouse embryos using a rapid, quantitative RT-PCR approach

Molecular Reproduction and Development
L RambhatlaKeith E Latham

Abstract

We have developed a novel reverse transcription-polymerase chain reaction (RT-PCR)-based approach for systematically quantifying in a single experiment the abundances of many different mRNAs in preimplantation mouse embryos. With this approach, the entire mRNA population from a small number of embryos is amplified while preserving the relative abundance of each mRNA in the cDNA population. The cDNA is analyzed by quantitative hybridization to radiolabeled probes. The approach is very sensitive and provides reliable, quantitative data regarding changes in mRNA abundance. A major advantage of this method is that estimates of mRNA copy number can be obtained and compared between different mRNAs. With this approach, we analyzed the patterns of expression of nine G protein alpha subunit mRNAs (G alpha s, G alpha i, G alpha q, G alpha o, and G alpha 11-15) in oocytes, eggs, and preimplantation embryos from fertilization to the blastocyst stage. Six alpha subunit mRNAs were expressed at significant levels, all of which underwent significant temporal alterations in expression. The mRNAs encoding some alpha subunit types were expressed predominantly in the egg and 1-cell embryo, underwent sharp reductions during the 2-cell stage, and we...Continue Reading

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Citations

Jul 1, 1996·Molecular Reproduction and Development·C J WilliamsG S Kopf
May 1, 1997·Molecular Reproduction and Development·A D DomashenkoK S Hatton
Jul 1, 1997·Molecular Reproduction and Development·Q Wang, K E Latham
Oct 10, 1997·Molecular Reproduction and Development·J V FlemingW D Rees
Oct 15, 1998·Molecular Reproduction and Development·A JurisicovaS L Varmuza
May 14, 1999·Developmental Genetics·K E LathamQ Wang
Feb 9, 2002·Molecular Reproduction and Development·Jean W WilliamsKeith E Latham
Jul 11, 2006·Apoptosis : an International Journal on Programmed Cell Death·Jacqui DetmarAndrea Jurisicova
Feb 24, 2012·Biology of Reproduction·Roxanne FernandesAndrea Jurisicova
Jun 6, 1998·Genes & Development·L BergeronJ Yuan
Jun 25, 2010·American Journal of Physiology. Cell Physiology·Santhi PotireddyKeith E Latham
Mar 12, 2009·American Journal of Physiology. Endocrinology and Metabolism·Jenna K Nyholt de PradaCatherine A VandeVoort
Jan 6, 2007·Human Cell·Toshio HamataniYasunori Yoshimura
Nov 29, 2005·Developmental Biology·Faical MiyaraKeith E Latham
Jun 7, 2007·Molecular Reproduction and Development·Rita VassenaKeith E Latham
Jul 23, 2008·Molecular Reproduction and Development·Namdori R MtangoKeith E Latham
Jan 1, 2010·Molecular Reproduction and Development·Jenna K Nyholt de PradaCatherine A Vandevoort
Aug 12, 2005·Molecular Reproduction and Development·Zohreh AlizadehFugaku Aoki
Sep 13, 2005·Molecular Reproduction and Development·Rita VassenaKeith E Latham
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