Rabbit Oocyte Cytoplasm Supports Development of Nuclear Transfer Embryos Derived from the Somatic Cells of the Camel and Tibetan Antelope

Abstract

Abstract. This study was designed to examine the ability of rabbit metaphase II oocyte cytoplasm to support the development of interspecies nuclear transfer embryos reconstructed using donor nuclei from different species. Skin fibroblast cells from a camel and Tibetan antelope were used as donor nuclei. As a first step, we investigated the efficiency of different activation protocols by comparing the parthenogenetic development of rabbit oocytes. The protocol that yielded the highest blastocyst rate was used to activate the reconstructed embryos in nuclear transfer experiments. In addition, the effect of donor cell serum starvation on the development of the reconstructed embryo was also examined. More than half of the karyoplast-cytoplast couplets could be fused, and about one third of the reconstructed embryos were capable of completing first cleavage, regardless of the species of donor nuclei. Some of the cleaving reconstructed embryos were even capable of progressing further and developing to the blastocyst stage (1.4-8.7% for the Tibetan antelope and 0-7.5% for the camel, respectively). Our results suggest that the mechanisms regulating early embryo development may be conserved among mammalian species and some factors existing in rabbit oocyte cytoplasm for somatic nucleus reprogramming and dedifferentiation may not be species-specific. Rabbit oocyte cytoplasm can reprogram donor nuclei regardless of the origin of the nucleus and support in vitro development to an advanced stage. [12]. At the same time, interspecies somatic cell cloned gaur and mouflon [13,14] have been obtained. Interspecies nuclear transfer, which involves transferring a donor cell from one species into a recipient oocyte of another species, is an invaluable tool for studying nucleuscytoplasm interaction and may be an effective way to conserve endangered species whose oocytes are extremely difficult and even impossible to obtain. Moreover, when interspecies nuclear transfer is used for therapeutic research, ethical, legal, and experimental limitations encountered in clinical situations may be avoided. One of the problems of interspecies nuclear transfer is the unavailability of species-specific competent recipient cytoplasm. In particular, it is extremely difficult to get oocytes for

Formation of nucleoli in interspecies nuclear transfer embryos derived from bovine, porcine, and rabbit oocytes and nuclear donor cells of various species

“…Together, with hypermethylation of porcine fibroblast nuclei , this may be the reason for failure of nucleoli formation in porcine-rabbit iSCNT embryos. However, rabbit oocytes were shown to support preimplantation development of embryos derived from nuclei of several species, including bovine , Capra ibex (Jiang et al 2005), chicken , camel and Tibetan antelope (Zhao et al 2006), macaca (Yang et al 2003), cat and panda (Wen et al 2005), and even human (Shi et al 2008). However, microarray analysis failed to detect significant human genome reprogramming in human-rabbit iSCNT embryos (Chung et al 2009).…”

“…The effect of IVC medium on iSCNT embryo development (Yang et al 2003, Zhao et al 2006, Sugimura et al 2009) is even more crucial, as it should support maternal RNA translation as well as EGA in xeno-ooplasm. It seems that medium efficiently supporting BL development of control SCNT embryos would sufficiently support the development of iSCNT embryos derived from the same oocytes (Zhao et al 2006). However, there might be also nuclear donor-species preference of IVC medium in spite of the same oocyte source (Yang et al 2003, Zhao et al 2006.…”

“…It seems that medium efficiently supporting BL development of control SCNT embryos would sufficiently support the development of iSCNT embryos derived from the same oocytes (Zhao et al 2006). However, there might be also nuclear donor-species preference of IVC medium in spite of the same oocyte source (Yang et al 2003, Zhao et al 2006. The data on the ability of IVC medium to support iSCNT embryo development are very contradictory.…”

Increased Th1/Th2 (IFN-.GAMMA./IL-4) Cytokine mRNA Ratio of Rat Embryos in the Pregnant Mouse Uterus

“…So, interspecific SCNT (iSCNT) has been explored to produce cloned offspring, and usually includes interspecific somatic cell nuclear transfer and interspecific pregnancy. In previous research, the oocytes of bovine, sheep, and rabbits have been used for iSCNT, and interspecific reconstructed embryos have been obtained [5][6][7][8][9][10][11][12][13]. Interspecific r e c o n s t r u c t e d e m b r y o s c a n i m p l a n t i n t o interspecific uteri, but only three speices have been successfully cloned by iSCNT [14][15][16].…”

Efecto De La Aplicación De Gonadotropinas Recombinantes Humanas Sobre La Producción Y Calidad De Ovocitos Y Embriones en Coneja

“…En muchas ocasiones la utilización del conejo como patrón de enfermedades humanas se basa en la utilización de poblaciones de animales transgénicos (Hirabayashi et al, 2000;Houdebine, 2005;Soler et al, 2005). En otras áreas de investigación el citoplasma de los ovocitos de coneja es empleado como receptor para la transferencia nuclear (Zhao et al, 2006;Chesne et al, 2002;Yang et al, 2004;Liu et al, 2005). Asimismo, también se emplea la técnica de inyección espermática intracitoplasmática, ICSI (Yamamoto et al, 1999;Ogonuki et al, 2005;Zhang et al, 2005), para todo lo cual se se requiere la utilización bien de embriones o bien de ovocitos.…”