Expression of the gap junction gene connexin43 (Cx43) in preimplantation bovine embryos derived in vitro or in vivo

Journal of Reproduction and Fertility
C WrenzyckiH Niemann

Abstract

In this study we have examined the presence of mRNA encoding connexin 43 (Cx43) in bovine embryos derived in vivo and in vitro. Cumulus-oocyte complexes, immature and matured oocytes liberated from cumulus cells, zygotes, 2-4-cell and 8-16-cell embryos, morulae, blastocysts and hatched blastocysts were produced in vitro from ovaries obtained from an abattoir using TCM 199 supplemented with hormones and 10% oestrous cow serum for maturation. Cumulus-oocyte complexes matured for 24 h were exposed to bull spermatozoa for 19 h and then cultured in TCM 199 supplemented with 10% oestrous cow serum to the desired developmental stage. Morulae and blastocysts derived in vivo were collected from superovulated donor cows. Total RNA was extracted from pools of 60-200 bovine oocytes or embryos using a modified phenol-chloroform extraction method and analysed by reverse transcriptase polymerase chain reaction. Before reverse transcription, aliquots of DNase-digested embryonic RNA were tested by polymerase chain reaction using bovine-specific primers to control for residual genomic DNA contamination. DNA-free, total RNA was reverse transcribed after preincubation with the Cx43 specific 3'primer. The resultant cDNA was amplified by polymerase ...Continue Reading

Citations

Jan 6, 2001·Molecular Reproduction and Development·A S LequarréA Van Langendonckt
Jan 30, 2004·Reproductive Biomedicine Online·Christine Wrenzycki, Heiner Niemann
Aug 25, 2009·The Journal of Reproduction and Development·Enrique GómezAlfonso Gutiérrez-Adán
Apr 29, 2008·The Journal of Reproduction and Development·Hitoshi UshijimaToshio Tajima
Jul 28, 2010·Reproduction : the Official Journal of the Society for the Study of Fertility·R WaurichK Jewgenow
Feb 8, 2005·Reproduction : the Official Journal of the Society for the Study of Fertility·Franchesca D Houghton
Nov 28, 2008·Reproduction : the Official Journal of the Society for the Study of Fertility·Enrique GómezJosé Néstor Caamaño
Mar 17, 2009·Molecular Reproduction and Development·M N PurperaK R Bondioli
Jul 15, 2004·Theriogenology·Deog-Bon KooYong-Mahn Han
Mar 15, 2008·Biochimica Et Biophysica Acta·Judith J Eckert, Tom P Fleming
May 21, 2013·Theriogenology·N N CostaO M Ohashi
Oct 31, 2007·Zygote : the Biology of Gametes and Early Embryos·H BadrR Puglisi

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