Serial nuclear transfer improves the development of interspecies reconstructed giant panda (Aluropoda melanoleuca) embryos

Interspecies somatic nuclear transfer (NT) may provide a new approach for preservation of the endangered rare species. Previous interspecies cloning studies have shown that a nucleus from a quiescent somatic cell supports early development of reconstructed embryos in the ooplasm from another species. In this study, we transferred nonquiescent somatic cells from a giant panda into the perivitelline space of the enucleated rabbit oocytes. After electrofusion (at the rate of 71.6％) and electrical activation, 4.2％ of the panda-rabbit reconstructed embryos developed to blastocyst in vitro. For improving the development rate of reconstructed embryos, we used serial NT in this study, I.e. Blastomeres from reconstructed morulae were transferred into the perivitelline space of the enucleated rabbit oocytes. The fusion rates in the groups of serial Ⅰ, serial Ⅱ and serial Ⅲ were 79.5％, 84.1％ and 78.0％, respectively, having no difference with that of somatic group. And the blastocyst rates in serial NT groups were 19.4％, 13.5％ and 10.3％, respectively, which are significantly higher than that in somatic NT group. These results indicate that the nuclei from nonquiescent somatic cells can support early development of reconstructed embryos and serial NT can improve the development rate of interspecies reconstructed embryos.     

Serial nuclear transfer improves the development of interspecies reconstructed giant panda (Aluropoda melanoleuca) embryos

Interspecies somatic nuclear transfer (NT) may provide a new approach for preservation of the endangered rare species. Previous interspecies cloning studies have shown that a nucleus from a quiescent somatic cell supports early development of reconstructed embryos in the ooplasm from another species. In this study, we transferred nonquiescent somatic cells from a giant panda into the perivitelline space of the enucleated rabbit oocytes. After electrofusion (at the rate of 71.6%) and electrical activation, 4.2% of the panda-rabbit reconstructed embryos developed to blastocyst in vitro. For improving the development rate of reconstructed embryos, we used serial NT in this study, i.e. blastomeres from reconstructed morulae were transferred into the perivitelline space of the enucleated rabbit oocytes. The fusion rates in the groups of serial I, serial II and serial III were 79.5%, 84.1% and 78.0%, respectively, having no difference with that of somatic group. And the blastocyst rates in serial NT groups were 19.4%, 13.5% and 10.3%, respectively, which are significantly higher than that in somatic NT group. These results indicate that the nuclei from nonquiescent somatic cells can support early development of reconstructed embryos and serial NT can improve the development rate of interspecies reconstructed embryos. These authors contributed equally to this work.     

Male panda （Ailuropoda melanoleuca） urine contains kinship information

Chemical communication plays an important role in kin selection and mate choice in mammals. The covariance of odor-genes of rodents has been documented and kinship odor has been proposed and termed, yet little is known of the relationship between genetic relatedness and chemical composition of kinship odors. Giant pandas （Ailuropoda melanoleuca） rely substantially on chemical communication to mediate their social interactions. To examine the relationship between genetic relatedness and compounds in the urine/anogenital gland secretions, we compared the similarities between genetic relatedness and the chemical profiles of anogenital gland secretions and urine via lineage construction and GC-MS （gas chromatography and mass spectrometry）. We found that information about kinship odors was present only in the urine of male adults in the mating season but absent in the non-mating season. Adult females and all sub-adults did not have such kinship odors in either mating or non-mating season. Therefore, kinship odor in the panda was contingent on age, sex, and season. This is the first report about the condition-dependent expression of kinship odor, which may have a significant implication in the practice of panda conservation in relation to chemical communication and sexual selection.