First deliveries of felines by transcervical transfer of in vitro-cultured embryos.

Transcervical transplantation of embryos has been wildly used in several species. However, it has not been successful in feline species because of their unique cervix structure combined with poor in vitro culture systems for embryos. In the present study, a simple device to guide the embryo transfer (ET) tube through the cervix and into the uterus was developed. To supply better blastocysts for ET, the in vitro culture system of feline embryos was also optimized. Results showed that 91.67 ± 4.17% of embryos developed to the blastocyst stage in QAU-4 medium compared with 49.17 ± 6.51% in KSOM medium. The total cell number of blastocysts reached 87.1 ± 22.1 in QAU-4 medium compared with 29.8 ± 2.1 in KSOM. Embryoid body formation was also observed in QAU-4 medium. Finally, by the improved technical scheme, CRX (Cone-Rod Homeobox) gene-edited kittens were successfully bred. To date, this is the first report, to our knowledge, of knockout kittens successfully delivered after transcervical ET with a simple transfer tube.

Differential dissolved protein expression throughout the life cycle of Giardia lamblia.

Giardia lamblia (G. lamblia) has a simple life cycle that alternates between a cyst and a trophozoite, and this parasite is an important human and animal pathogen. To increase our understanding of the molecular basis of the G. lamblia encystment, we have analyzed the soluble proteins expressed by trophozoites and cysts extracted from feces by quantitative proteomic analysis. A total of 63 proteins were identified by isobaric tags for relative and absolute quantitation (iTRAQ) labeling, and were categorized as cytoskeletal proteins, a cell-cycle-specific kinase, metabolic enzymes and stress resistance proteins. Importantly, we demonstrated that the expression of seven proteins differed significantly between trophozoites and cysts. In cysts, the expression of three proteins (one variable surface protein (VSP), ornithine carbamoyltransferase (OTC), ß-tubulin) increased, whereas the expression of four proteins (14-3-3 protein, α-tubulin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), protein disulfide isomerase 2 (PDI-2)) decreased significantly when compared with the levels of these proteins in trophozoites. The mRNA expression patterns of four of these proteins (OTC, α-tubulin, GAPDH, VSP) were similar to the expression levels of the proteins. These seven proteins appear to play an important role in the completion of the life cycle of G. lamblia.