Effects of Time of Deoxyribonucleic Acid Microinjection on Gene Detection and In Vitro Development of Bovine Embryos

Authors

A.E T. Sparks, Department of Dairy Science, Virginia Polytechnic Institute and State University, Blacksburg 24061 -0315
R S. Canseco, Department of Dairy Science, Virginia Polytechnic Institute and State University, Blacksburg 24061 -0315
C G. Russell, Department of Chemical Engineering, Virginia Polytechnic Institute and State University, Blacksburg 24061 -0315
J L. Johnson, Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg 24061 -0315
H D. Moll, Virginia /Maryland Regional College of Veterinary Medicine
William H. Velander, University of Nebraska-LincolnFollow
Francis C. Gwazdauskas, Deparment of Dairy Science, Virginia Polytechnic Institute and State University, Blacksburg, VAFollow

Date of this Version

March 1994

Comments

Originally Published in Journal of Dairy Science, March 1994 Vol 77 No.3. This article can be viewed at: http://jds.fass.org/cgi/reprint/77/3/718

Abstract

In vivo fertilized embryos were surgically collected from superovulated dairy cows to evaluate microinjection on embryo development and utilized the polymerase chain reaction technique for selection of transgenic embryos. Seventy-two percent of the embryos with visible pronuclei or nuclei were microinjected with DNA, and the remaining 28% served as uninjected controls. All embryos were cocultured with bovine oviductal epithelial cells. Mean final development scores of embryos within the same initial cell stage at collection were unaffected by microinjection. After 144 h of culture, 45% of the microinjected embryos developed to the morula or blastocyst stage. The transgene was detected in 50, 10, and 9% of demimorulae from embryos microinjected at the 1-, 2-, and 4-cell stages. Frequency of transgene detection was higher in morulae from 1-cell embryos than in morulae from 2- and 4-cell embryos. Use of in vitro coculture, embryo bisection, and polymerase chain reaction technique facilitated selection of bovine embryos that carried the transgene.