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Titolo:
DEVELOPMENT OF PARTHENOGENETIC AND CLONED OVINE EMBRYOS - EFFECT OF ACTIVATION PROTOCOLS
Autore:
LOI P; LEDDA S; FULKA J; CAPPAI P; MOOR RM;
Indirizzi:
IST ZOOTECN & CASEARIO SARDEGNA I-07040 OLMEDO ITALY DIPARTIMENTO BIOL ANIM SASSARI ITALY INST ANIM PROD PRAGUE CZECH REPUBLIC BABRAHAM INST,DEPT GENET & DEV CAMBRIDGE CB2 4AT ENGLAND
Titolo Testata:
Biology of reproduction
fascicolo: 5, volume: 58, anno: 1998,
pagine: 1177 - 1187
SICI:
0006-3363(1998)58:5<1177:DOPACO>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
MOUSE EGG ACTIVATION; NUCLEAR TRANSPLANTATION; CELL-CYCLE; BOVINE OOCYTES; 6-DIMETHYLAMINOPURINE 6-DMAP; PROTEIN-KINASES; MAMMALIAN EGGS; SHEEP EMBRYOS; CALCIUM; FERTILIZATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
58
Recensione:
Indirizzi per estratti:
Citazione:
P. Loi et al., "DEVELOPMENT OF PARTHENOGENETIC AND CLONED OVINE EMBRYOS - EFFECT OF ACTIVATION PROTOCOLS", Biology of reproduction, 58(5), 1998, pp. 1177-1187

Abstract

Preliminary experiments carried out on ovine oocytes were designed toestablish correlations between activation protocols and subsequent rates of embryonic development. The best activation protocols were thereafter used in studies on ovine parthenogenesis and cloning. The first study established that chemical activators induce pronuclear development at a slightly higher rate than physical activation (ionomycin, 96%;ethanol, 95%; electro activation, 80%). Inhibition of second polar body extrusion and one single pronucleus were observed in the majority of the oocytes (similar to 90%) treated for 3 h with 6-dimethylaminopurine (6-DMAP) following either ionomycin or ethanol activation. While over 80% of these oocytes cleaved after transfer to the oviducts of recipients, progression to the blastocyst stage was higher after ionomycin as compared with ethanol activation (58% vs. 19%). The ionomycin plus 6-DMAP activation protocol was used to produce parthenogenetic blastocysts whose subsequent development was monitored both by ultrasonography and by direct fetal examination. Over 70% of parthenogenotes were viable on Day 21 of pregnancy but dead by Day 25. The effects of 6-DMAP on nuclear remodeling and fetal development of cloned embryos was then investigated. Control cloned embryos underwent nuclear envelope breakdown (NEBD), premature chromatin condensation (PCC), and inhibition of DNA synthesis. By contrast, reconstructed embryos treated with 6-DMAP exhibited intact nuclear membranes, interphase chromatin, and no interference on DNA synthesis. Moreover, cloned embryos developed to blastocyst;stage in higher percentage after 6-DMAP treatment (83% vs. 25%). We conclude that ionomycin followed by 6-DMAP incubation yields high percentages of diploid parthenogenetic embryos that develop to Day 25 before dying. Cloned embryos activated by the ionomycin-6-DMAP protocol develop readily to term.

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Documento generato il 04/12/20 alle ore 03:06:33