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Titolo:
EFFECT OF MICROINJECTION TIME DURING POSTFERTILIZATION S-PHASE ON BOVINE EMBRYONIC-DEVELOPMENT
Autore:
GAGNE M; POTHIER F; SIRARD MA;
Indirizzi:
CHU LAVAL,UNITE RECH ONTOGENIE & REPROD,2705 BOUL LAURIER ST FOY PQ G1V 4G2 CANADA CHU LAVAL,UNITE RECH ONTOGENIE & REPROD ST FOY PQ G1V 4G2 CANADA UNIV LAVAL,DEPT ANIM SCI ST FOY PQ G1K 7P4 CANADA
Titolo Testata:
Molecular reproduction and development
fascicolo: 2, volume: 41, anno: 1995,
pagine: 184 - 194
SICI:
1040-452X(1995)41:2<184:EOMTDP>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
DNA-POLYMERASE-ALPHA; GERMINAL VESICLE BREAKDOWN; CELL NUCLEAR ANTIGEN; SEA-URCHIN EGGS; PROTEIN-SYNTHESIS; MOUSE OOCYTES; CHROMOSOMAL INTEGRATION; MEIOTIC MATURATION; RABBIT OVIDUCTS; CULTURED-CELLS;
Keywords:
MICROINJECTION; S-PHASE; ZYGOTES; BOVINE; DEVELOPMENT;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
82
Recensione:
Indirizzi per estratti:
Citazione:
M. Gagne et al., "EFFECT OF MICROINJECTION TIME DURING POSTFERTILIZATION S-PHASE ON BOVINE EMBRYONIC-DEVELOPMENT", Molecular reproduction and development, 41(2), 1995, pp. 184-194

Abstract

Microinjection into bovine zygotes was performed to evaluate the effects of the timing of injection during the phase of DNA replication on the subsequent in vitro development of embryos and expression of injected chicken beta-actin promoter-lac Z gene construct. The period of DNA replication of bovine zygotes, determined by H-3-thymidine incorporation, begins between 12 hr and 13 hr postinsemination (hpi) of in vitro matured oocytes, reaches a maximum from 17 hpi to 19 hpi, and is complete by 21-22 hpi. Aphidicolin, an inhibitor of DNA polymerase alpha,was used to synchronize the pronuclei and the zygote population. Treatment with aphidicolin at 9-18 hpi arrested DNA replication without affecting formation of the pronuclei or embryo development. Cycloheximide, an inhibitor of protein synthesis, was used for nucleocyloplasmic resynchronization of the aphidicolin-treated zygotes. Microinjection was performed at 15 (early), 18 (mid), and 21 (late S phase) hpi. Embryonic development was affected following each of the three microinjection times. The development of zygotes injected at 18 hpi was significantly higher (P < 0.01) after 5 days of culture than those injected at 15hpi or 21 hpi. Expression of the marker gene was observed in the higher stage of development (>16 cells) only in the zygotes injected at 18hpi. At the earlier stages of development, the proportions of embryosshowing expression of the foreign gene were the same for all microinjection times. In aphidicolin- and cycloheximide-treated zygotes, expression of the marker gene followed the same curve as development, i.e.,expression was low when injected early or late and higher (P < 0.005)when injected in the middle of zygotic S phase. The ability of the embryos to survive microinjection and to express the marker gene as a function of hpi seems to be influenced mostly in the cytoplasm processing stage rather than the pronuclei processing stage. (C) 1995 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/11/20 alle ore 10:04:15