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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2207-2211 |
| Number of pages | 5 |
| Journal | Journal of animal science |
| Volume | 70 |
| Issue number | 7 |
| DOIs | |
| State | Published - Jul 1992 |
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ASJC Scopus subject areas
- Food Science
- Animal Science and Zoology
- Genetics
Cite this
In vitro development of zygotes from prepubertal gilts after microinjection of DNA. / Williams, B. L.; Sparks, A. E.; Canseco, R. S.; Knight, J. W.; Johnson, J. L.; Velander, W. H.; Page, R. L.; Drohan, W. N.; Young, J. M.; Pearson, R. E.
In: Journal of animal science, Vol. 70, No. 7, 07.1992, p. 2207-2211.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - In vitro development of zygotes from prepubertal gilts after microinjection of DNA.
AU - Williams, B. L.
AU - Sparks, A. E.
AU - Canseco, R. S.
AU - Knight, J. W.
AU - Johnson, J. L.
AU - Velander, W. H.
AU - Page, R. L.
AU - Drohan, W. N.
AU - Young, J. M.
AU - Pearson, R. E.
PY - 1992/7
Y1 - 1992/7
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0026887197&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026887197&partnerID=8YFLogxK
U2 - 10.2527/1992.7072207x
DO - 10.2527/1992.7072207x
M3 - Article
C2 - 1644695
AN - SCOPUS:0026887197
VL - 70
SP - 2207
EP - 2211
JO - Journal of Animal Science
JF - Journal of Animal Science
SN - 0021-8812
IS - 7
ER -