In vitro development of zygotes from prepubertal gilts after microinjection of DNA

B L Williams; A E Sparks; R S Canseco; J W Knight; J L Johnson; W H Velander; R L Page; W N Drohan; J M Young; R E Pearson

doi:10.2527/1992.7072207x

Back

In vitro development of zygotes from prepubertal gilts after microinjection of DNA

Journal article

Peer reviewed

In vitro development of zygotes from prepubertal gilts after microinjection of DNA

B L Williams, A E Sparks, R S Canseco, J W Knight, J L Johnson, W H Velander, R L Page, W N Drohan, J M Young and R E Pearson

Journal of animal science, Vol.70(7), pp.2207-2211

07/1992

DOI: 10.2527/1992.7072207x

PMID: 1644695

View Online

Abstract

The effect of pronuclear microinjection of DNA and culture in excised mouse oviducts on the development of porcine zygotes was assessed in this study. Precocious ovulation was induced in prepubertal gilts with pregnant mare's serum gonadotrophin and hCG. Zygotes received either pronuclear microinjection of buffer alone, buffer containing a DNA construct, or no microinjection. Zygotes were cultured in vitro in either modified Krebs-Ringer bicarbonate medium (KRB) for 144 h or in mouse oviduct (MO) explant culture with KRB for 48, 72, 96, or 120 h. Pronuclear microinjection of DNA resulted in a lower (P less than .05) cleavage index (CI) than did buffer or no microinjection (CI 2.16 +/- .10 vs 2.80 +/- .13 and 2.93 +/- .10). The CI loss was greatest for DNA-injected zygotes at the two-cell stage of development. Coculture of zygotes in MO resulted in a higher CI (P less than .01) than did culture in KRB. Culture in MO for 72 h was the most beneficial system compared with MO for 48, 96, or 120 h (P less than .05; CI 3.25 +/- .12 vs 2.66 +/- .18, 2.79 +/- .14, and 2.40 +/- .14, respectively). Microinjection of DNA, not merely the mechanical procedure, was detrimental to early zygote development and may be the cause of low pregnancy rates.

Microinjections

Culture Techniques

DNA - pharmacology

Cells, Cultured

Fallopian Tubes

Swine - physiology

Animals

Culture Media

Female

Mice

Ovulation Induction - veterinary

Zygote - growth & development

Sexual Maturation

Details

Title: Subtitle: In vitro development of zygotes from prepubertal gilts after microinjection of DNA
Creators: B L Williams - Department of Dairy Science, College of Agriculture and Life Science, Virginia Agricultural Experiment Station, Blacksburg 24061
A E Sparks
R S Canseco
J W Knight
J L Johnson
W H Velander
R L Page
W N Drohan
J M Young
R E Pearson
Resource Type: Journal article
Publication Details: Journal of animal science, Vol.70(7), pp.2207-2211
Publisher: United States
DOI: 10.2527/1992.7072207x
PMID: 1644695
ISSN: 0021-8812
eISSN: 1525-3163
Language: English
Date published: 07/1992
Academic Unit: Obstetrics and Gynecology
Record Identifier: 9983930385102771

Metrics

19 Record Views

18 Times Cited - Web of Science

See more details