체외생산된 소 배반포강 내의 아미노산 농도

Amino Acid Concentrations in the Blastocoelic Fluid of In Vitro-Produced Bovine Blastocysts

체외생산된 소 배반포 및 탈출 배반포강 내의 유리 아미노산 농도를 측정하였다. 체외 배양은 소 혈청알부민이 함유된 합성난관배양액(synthetic oviduct fluid; SOF)에서 실시하였으며 수정 후 180시간의 배반포 및 216시간 후의 탈출 배반포를 실험에 공여하였다 아미노산 측정은 알부민 대신 Polyvinyl alcohol이 함유된 SOF 미소적에 수정란을 분주하고 미세조작기를 이용하여 배반포강 내의 액을 추출하여 20종류의 아미노산을 측정하였다. 탈출 배반포는 isoleucine, leucine 및 methionine 농도가 배반포보다 유의적(p〈0.05)으로 높게 나타났으며, glutamate, aspartate는 두 군간에 차이를 나타내지 알았다. 반면 alanine 및 threamine (p〈0.01)과 cystine을 제외한 나머지 12종류의 아미노산(p〈0.001)은 배반포가 탈출 배반포에 비해서 유의적으로 높은 측정치를 나타내었다. 비록 glutamine의 경우 배양액 내에 첨가되지 않았으나 두 군, 특히 배반포 내에서 높은 측정치를 보였다. 본 연구결과로 보아 체외생산된 배반포 및 탈출 배반포는 내강에 필수 및 비필수 아미노산을 각기 다른 농도로 함유하고 있는 것으로 사료된다.

Concentrations of free amino acids in the BF of IVP bovine BL and HBL were examined in this study. The embryos derived from IVF oocytes were cultured in a SOFM containing BSA, EAA and NEAA. BF was aspirated from BL (180 h of age after insemination) and HBL (216 h of age after insemination), and introduced into drops of SOFM (30$\mu$l/drop) containing PVA through micromanipulation. The medium containing BF was then subjected to measurement of 20 amino acids by an automatic amino acid analyzer. The concentrations of isoleucine, leucine and methionine were higher (p〈0.05) in the BF from HBL than from BL, and no difference was found in aspartate or glutamate concentrations between BL and HBL, while threonine, alanine (p〈0.01) and the rest of the amino acids (p〈0.001) were significantly higher in the BF from HBL than from BL. Cystine was not found in either BL or HBL. A high concentration of glutamine was found in the BF from both BL and HBL, although it was not added to the culture medium. These results indicate that bovine BF contains several EAA (methionine in BL and isoleucine, leucine and methionine in HBL) and NEAA (alanine, glutamate, glycine, proline, serine and aspartate in BL, and glutamate and aspartate in HBL), and there is significant differences in the amino acid concentration in the BF between BL and HBL derived by WP.