Skip to main content
Article
Effects of time of deoxyribonucleic acid microinjection on gene detection and in vitro development of bovine embryos
Journal of dairy science
  • Amy E.T. Sparks, University of Iowa
  • R. S. Canseco
  • C. G. Russell
  • J. L. Johnson
  • H. D. Moll
  • W. H. Velander
  • F. C. Gwazdauskas
Document Type
Article
Peer Reviewed
1
Publication Date
3-1-1994
NLM Title Abbreviation
J Dairy Sci
PubMed ID
8169279
DOI of Published Version
10.3168/jds.S0022-0302(94)77005-3
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.

Keywords
  • Animals,
  • Animals,
  • Genetically Modified/embryology,
  • Base Sequence,
  • Blastocyst/physiology,
  • Cattle,
  • Culture Techniques,
  • DNA/administration & dosage,
  • Embryonic and Fetal Development,
  • Female,
  • Gene Transfer Techniques,
  • Microinjections,
  • Molecular Sequence Data,
  • Morula/physiology,
  • Polymerase Chain Reaction,
  • Superovulation,
  • Time Factors
Published Article/Book Citation
Journal of dairy science, 77:3 (1994) pp.718-724.
Citation Information
Amy E.T. Sparks, R. S. Canseco, C. G. Russell, J. L. Johnson, et al.. "Effects of time of deoxyribonucleic acid microinjection on gene detection and in vitro development of bovine embryos" Journal of dairy science Vol. 77 Iss. 3 (1994) p. 718 - 724 ISSN: 0022-0302
Available at: http://works.bepress.com/amy_sparks/59/