Development, embryonic genome activity and mitochondrial characteristics of bovine-pig inter-family nuclear transfer embryos.
The best results of inter-species somatic cell nuclear transfer (iSCNT) in mammals were obtained using closely related species that can hybridise naturally. However, in the last years, many reports describing blastocyst development following iSCNT between species with distant taxonomical relations (inter-classes, inter-order and inter-family) have been published. This indicates that embryonic genome activation (EGA) in xeno-cytoplasm is possible, albeit very rarely. Using a bovine-pig (inter-family) iSCNT model, we studied the basic characteristics of EGA: expression and activity of RNA polymerase II (RNA Pol II), formation of nucleoli (as an indicator of RNA polymerase I (RNA Pol I) activity), expression of the key pluripotency gene NANOG and alteration of mitochondrial mass. In control embryos (obtained by IVF or iSCNT), EGA was characterised by RNA Pol II accumulation and massive production of poly-adenylated transcripts (detected with oligo dT probes) in blastomere nuclei, and formation of nucleoli as a result of RNA Pol I activity. Conversely, iSCNT embryos were characterised by the absence of accumulation and low activity of RNA Pol II and inability to form active mature nucleoli. Moreover, in iSCNT embryos, NANOG was not expressed, and mitochondria mass was significantly lower than in intra-species embryos. Finally, the complete developmental block at the 16-25-cell stage for pig-bovine iSCNT embryos and at the four-cell stage for bovine-pig iSCNT embryos strongly suggests that EGA is not taking place in iSCNT embryos. Thus, our experiments clearly demonstrate poor nucleus-cytoplasm compatibility between these animal species.
Laboratorio di Tecnologie della Riproduzione, Avantea srl, Via Porcellasco 7/f, Cremona, Italy. irinalagutina@avantea.it
Irina
I
Lagutina
Helena
H
Fulka
Tiziana A L
TA
Brevini
Stefania
S
Antonini
Dario
D
Brunetti
Silvia
S
Colleoni
Fulvio
F
Gandolfi
Giovanna
G
Lazzari
Josef
J
Fulka
Cesare
C
Galli
D000818:Animals; D002417:Cattle / Q000235:genetics / Q000502:physiology*; D002467:Cell Nucleus / Q000502:physiology; D024142:Collagen Type VI / Q000235:genetics / Q000502:physiology; D003593:Cytoplasm / Q000502:physiology; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004622:Embryo, Mammalian / Q000502:physiology*; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D008297:Male; D008928:Mitochondria / Q000502:physiology; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011247:Pregnancy; D012318:RNA Polymerase I / Q000235:genetics / Q000502:physiology; D012319:RNA Polymerase II / Q000235:genetics / Q000502:physiology; D013552:Swine / Q000235:genetics / Q000502:physiology*
D024142:Collagen Type VI; D004247:DNA; D012319:RNA Polymerase II; D012318:RNA Polymerase I