Mdivi-1, mitochondrial fission inhibitor,impairs developmental competence and mitochondrial function of embryos and cells in pigs

Mitochondria are highly dynamic organelles that undergo constant fusion/fission as well as activities orchestrated by large dynamin-related GTPases. These dynamic mitochondrial processes influence mitochondrial morphology, size and function. Therefore, this study was conducted to evaluate the effects of mitochondrial fission inhibitor, mdivi-1, on developmental competence and mitochondrial function of porcine embryos and primary cells. Presumptive porcine embryos were cultured in PZM-3 medium supplemented with mdivi-1 (0, 10 and 50 μM) for 6 days. Porcine fibroblast cells were cultured in growth medium with mdivi-1 (0 and 50 μM) for 2 days. Our results showed that the rate of blastocyst production and cell growth in the mdivi-1 (50 μM) treated group was lower than that of the control group (P < 0.05). Moreover, loss of mitochondrial membrane potential in the mdivi-1 (50 μM) treated group was increased relative to the control group (P < 0.05). Subsequent evaluation revealed that the intracellular levels of reactive oxygen species (ROS) and the apoptotic index were increased by mdivi-1 (50 μM) treatment (P < 0.05). Finally, the expression of mitochondrial fission-related protein (Drp 1) was lower in the embryos and cells in the mdivi-1-treated group than the control group. Taken together, these results indicate that mdivi-1 treatment may inhibit developmental competence and mitochondrial function in porcine embryos and primary cells.     

A Cathepsin B Inhibitor,E-64, Improves the Preimplantation Development of Bovine Somatic Cell Nuclear Transfer Embryos

Bovine somatic cell nuclear transfer (SCNT) is an important and powerful tool for basic research and biomedical and agricultural applications, however, the efficiency of SCNT has remained extremely low. In this study, we investigated the effects of cathepsin B inhibitor (E-64) supplementation of culture medium on in vitro development of bovine SCNT embryos. We initially used three concentrations of E-64 (0.1, 0.5, 1.0 μm), among which 0.5 μm resulted in the highest rate of blastocysts production after in vitro fertilization (IVF), and was therefore used for further experiments. Blastocyst development of SCNT embryos in the E-64 treatment group also increased relative to the control. Moreover, the cryosurvival rates of IVF and SCNT blastocysts were increased in E-64 treatment groups when compared with the control. On the other hand, we found that IVF and SCNT blastocysts derived from E-64-treated groups had increased total cell numbers and decreased apoptotic nuclei. Furthermore, assessment of the expression of apoptosis-related genes (Bax and Bcl-xL) in bovine IVF and SCNT blastocysts treated with E-64 by real-time RT-PCR analysis revealed suppressed expression of the pro-apoptotic gene Bax and stimulated expression of the anti-apoptotic gene Bcl-xL. Taken together, these finding indicate that addition of E-64 to embryo culture medium may have important implications for improving developmental competence and preimplantation quality in bovine IVF and SCNT embryos.     

Imaging characteristics of the eyes of cinereous vulture (Aegypius monachus): morphology and comparative biometric measurement

The aim of this study is to describe radiographic, ultrasonographic, and computed tomographic appearance of normal cinereous vulture’s eye and to determine normal biometric values of intraocular structures. Twenty-six eyes of thirteen healthy cinereous vultures were examined. Under general anesthesia with isoflurane, ultrasonography (US), computed tomography (CT) and skull radiography were performed. Differences between both eyes as well as between US and CT measurements were investigated and correlation of measurements between both eyes as well as correlation between CT and US measurements of the various ocular structures were calculated. Most of paired data did not show any significant differences between both eyes and the CT and US measurements, while there were significant differences (P<0.05) between CT and US measurements of depth of both vitreous and anterior chambers, and axial length of the lens in right eyes. There was also a significant difference (P<0.05) between both eyes in depth of vitreous measured by CT. All the measurements had strong correlations between both eyes and between US and CT. In conclusion, ocular imaging techniques provided useful data of biometry and morphology, showing good correlation between CT and US in cinereous vulture’s eye. Especially, when ophthalmoscopic examinations would not be available due to opaque anterior segment, imaging techniques could be essential for diagnosing and managing of the eye.     

Severe hydrocephalus in a raccoon dog (Nyctereutes procyonoides)

Hydrocephalus is one of the most common central nervous system malformations in domestic dogs, yet they are poorly documented and studied in wild carnivoran mammals. A pup of raccoon dog (Nyctereutes procyonoides) was rescued and brought to Wildlife Center. The pup showed generalized ataxia, a domed skull, and an open bregmatic fontanelle. Ultrasound and MRI showed severe enlargement of the lateral ventricle with the loss of septum pellucidum resulting in a single large ventricle and cervical syringohydromyelia. Although treatment was attempted, the animal was euthanized due to poor prognosis. At necropsy, macroscopic findings were identical to the diagnostic imaging, where marked enlargement of the calvarium, and attenuated gyri and sulci were observed. Finally, hydrocephalus was confirmed. Here, we describe a case of hydrocephalus in a raccoon dog (Nyctereutes procyonoides).