Technical, biological and molecular aspects of somatic cell nuclear transfer – a review

Since the announcement of the birth of the first cloned mammal in 1997, Dolly the Sheep, 24 animal species including laboratory, farm, and wild animals have been cloned. The technique for somatic cloning involves transfer of the donor nucleus of a somatic cell into an enucleated oocyte at the metaphase II (MII) stage for the generation of a new individual, genetically identical to the somatic cell donor. There is increasing interest in animal cloning for different purposes such as rescue of endangered animals, replication of superior farm animals, production of genetically engineered animals, creation of biomedical models, and basic research. However, the efficiency of cloning remains relatively low. High abortion, embryonic, and fetal mortality rates are frequently observed. Moreover, aberrant developmental patterns during or after birth are reported. Researchers attribute these abnormal phenotypes mainly to incomplete nuclear remodeling, resulting in incomplete reprogramming. Nevertheless, multiple factors influence the success of each step of the somatic cloning process. Various strategies have been used to improve the efficiency of nuclear transfer and most of the phenotypically normal born clones can survive, grow, and reproduce. This paper will present some technical, biological, and molecular aspects of somatic cloning, along with remarkable achievements and current improvements.

The Roles of the miRNAome and Transcriptome in the Ovine Ovary Reveal Poor Efficiency in Juvenile Superovulation

Juvenile superovulation can provide a wealth of oocyte material for embryo production, animal cloning, and genetic modification research, but embryos derived from juvenile oocytes show poor efficiency in subsequent developmental capacity. In order to reveal the formation mechanism of large numbers of follicles and poor oocyte quality in juvenile ovaries under superovulation treatment, differentially expressed microRNAs (miRNAs) and messenger RNAs (mRNAs) were characterized and investigated in the ovaries of lambs and adult sheep using high-throughput sequencing technology. The majority of differentially expressed miRNAs (337/358) were upregulated in lamb libraries. The expression levels of mRNAs related to hormone receptors (follicle-stimulating hormone receptor, FSHR; luteinizing hormone/choriogonadotropin receptor, LHCGR; estrogen receptor 1, ESR1), steroid hormone secretion (cytochrome P450 family 11 subfamily A member 1, CYP11A1; cytochrome P450 family 17 subfamily A member 1, CYP17A1; cytochrome P450 family 19 subfamily A member 1, CYP19A1), and oocyte quality (pentraxin 3, PTX3; BCL2 apoptosis regulator, BCL2; caspase 3, CASP3) were significantly different between the lamb and adult libraries. The miRNA aor-miR-143, which targets FSHR, was highly and differentially expressed, and PTX3 was predicted to be targeted by oar-miR-485-3p and oar-miR-377-3p in the ovine ovary. A considerable number of miRNAs were predicted to inhibit ESR1 expression in lamb ovaries. In conclusion, oar-miR-143 and FSHR molecules, among others, might regulate follicle formation, and oar-miR-485-3p, oar-miR-377-3p, and PTX3, among others, may be associated with oocyte quality. These identified miRNAs and mRNAs will be beneficial for the prediction of ovarian superovulation potential and screening of oocytes.

Impact of Socioscientific Issues on Middle School Students' Character and Values for Global Citizenship

This chapter demonstrates the impact of an animal cloning socioscientific issues instructional unit on a group of middle school students' abilities to use their ecological worldview, social and moral compassion, and sense of socioscientific accountability to determine the permissibility of animal cloning. Seventy-seven 7th grade students at a public middle school in the Southeastern region of the United States participated in this investigation. Results from a non-parametric two-tailed Wilcoxon test indicated the students' social and moral compassion (Z = -2.505, p = .012) and socioscientific accountability scores (Z = -2.381, p = .017). In contrast, the results did not demonstrate a statistically significant difference between students' pre and post ecological worldview (Z = -1.185, p = .236). Qualitative analyses of the data revealed several interesting trends and themes discussed in the chapter. The findings from this investigation support the use of SSI as key pedagogical strategies in promoting character and values for global citizens among middle school students.

Pyometra does not affect some molecular quality-related parameters of canine oocytes

Cystic endometrial hyperplasia-pyometra complex (CEH/P) significantly perturbs the reproductive performance of affected bitches and ovariohysterectomy (OHE) is a commonly applied treatment. Thus the only way to take advantage of the genetic potential of valuable females is application of assisted reproductive techniques (ART) mainly in vitro embryo production (IVP) or in some exceptional cases animal cloning by somatic cell nuclear transfer (SCNT). The aim of our study was to examine a potential effect of the CEH/P status on the quality of oocytes from females subjected to OHE. In total, 828 immature oocytes collected from ovaries of 33 bitches (21 control, 12 CEH/P) were subjected to genetic analyses (mRNA expression of two maternal-effect genes: GDF-9, OCT4 and mitochondrial DNA – mtDNA - content). Oocytes of CEH/P females were characterized by a higher mtDNA content (471 696) than gametes of their healthy counterparts (368 175; p<0,005). Transcripts for the two genes were detected in all samples and the mRNA level was not affected by the CEH/P status. In conclusion, the CEH/P complex does not exert a negative effect on oocyte quality reflected by the two parameters examined in this study.