Hwanyul Yong

Effect of exogenous carbohydrates in a serum-free culture medium on the development of in vitro matured and fertilized porcine embryos

This study was conducted to examine the effect of energy substrates in a serum-free culture medium on in vitro development of porcine embryos. Presumptive zygotes derived from in vitro fertilization were cultured in glucose-free North Carolina State University (NCSU)-23 medium with glucose, pyruvate, fructose and lactate added to the culture medium singly or in various combinations. In experiment 1, a higher percentage of embryos cleaved (53-63% vs 10-13%) and developed to the blastocyst stage (18-27% vs 0) after the single addition of glucose (5.6 mM), pyruvate (0.5 mM) or lactate (10 mM) than with no energy substrate addition or the addition only of fructose (5.6 mM). In experiment 2, the addition of pyruvate and lactate resulted in higher blastocyst formation (25%) than other combinations (6-22%), while the addition of glucose and pyruvate significantly inhibited blastocyst formation. Increasing lactate concentration, as a single energy supplement, from 5 to 20 mM significantly improved blastocyst formation (7% vs 14-18%), while no benefit was achieved from increasing pyruvate concentration up to 2 mM (experiment 3). Glucose-free NCSU-23 medium supplemented with 0.5 mM pyruvate and 5 mM lactate significantly improved blastocyst formation (28% vs 17%) compared with NCSU-23 medium supplemented with 5.6 mM glucose (experiment 4). In conclusion, pyruvate and lactate are preferable energy substrates to support in vitro development of porcine embryos cultured in a serum-free NCSU-23 medium.

Rapamycin treatment during in vitro maturation of oocytes improves embryonic development after parthenogenesis and somatic cell nuclear transfer in pigs

This study was conducted to investigate the effects of rapamycin treatment during in vitro maturation (IVM) on oocyte maturation and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) in pigs. Morphologically good (MGCOCs) and poor oocytes (MPCOCs) were untreated or treated with 1 nM rapamycin during 0-22 h, 22-42 h, or 0-42 h of IVM. Rapamycin had no significant effects on nuclear maturation and blastocyst formation after PA of MGCOCs. Blastocyst formation after PA was significantly increased by rapamycin treatment during 22-42 h and 0-42 h (46.6% and 46.5%, respectively) relative to the control (33.3%) and 0-22 h groups (38.6%) in MPCOCs. In SCNT, blastocyst formation tended to increase in MPCOCs treated with rapamycin during 0-42 h of IVM relative to untreated oocytes (20.3% vs. 14.3%, 0.05 < p < 0.1), while no improvement was observed in MGCOCs. Gene expression analysis revealed that transcript abundance of Beclin 1 and microtubule-associated protein 1 light chain 3 mRNAs was significantly increased in MPCOCs by rapamycin relative to the control. Our results demonstrated that autophagy induction by rapamycin during IVM improved developmental competence of oocytes derived from MPCOCs.

Use of a domestic Korean black goat (Capra hircus coreanae) with its chest crayon-harnessed in detecting estrus of Himalayan tahrs (Hemitragus jemlahicus)

The reliability of a Korean black goat (Capra hircus coreanae) to detect estrus in Himalayan tahrs (Hemitragus jemlahicus) for an artificial breeding program was investigated. Estrus in six female Himalayan tahrs was synchronized using fluorogestone acetate (FGA) sponges. Thirteen days later, 200 IU of PMSG and 100 IU of hCG were injected before removing the sponges and simultaneously injecting 5 mg of PGF2α the next day. Penetration of the cervical canal and the thickness and location of red crayon marks were examined 40~43 h later. Two females treated with sponges containing 60 or 45 mg of FGA had estrogen levels of 8.7 and 11.1 pg/mL, respectively. No red marks were found on the backs of these two tahrs. The remaining females had higher levels of estradiol, and the red crayon marks were clearly shown. The cervical folds of these tahrs were readily penetrated and the insemination gun was smoothly inserted into the uterine body. In conclusion, a Korean domestic goat with its chest crayon-harnessed was successfully used to detect estrus of Himalayan tahrs. This technique might be utilized as a part of breeding programs for wild goats and avoid the need for a vasectomy of conspecific males.