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.
Department of Biotechnology, College of Engineering, Daegu University, Gyeongbuk 712-714, Republic of Korea.
Sung-Hun
SH
Min
Bong-Seok
BS
Song
Ji-Yeong
JY
Yeon
Jin-Woo
JW
Kim
Jung-Ho
JH
Bae
Soo-Yong
SY
Park
Yong-Hee
YH
Lee
Kyu-Tae
KT
Chang
Deog-Bon
DB
Koo
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002401:Cathepsin B / Q000037:antagonists & inhibitors*; D002417:Cattle; D015853:Cysteine Proteinase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D010064:Embryo Implantation; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D007930:Leucine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D053652:Nuclear Transfer Techniques / Q000662:veterinary*
D015853:Cysteine Proteinase Inhibitors; D002401:Cathepsin B; D007930:Leucine; C024974:E 64