Practical Applications of DNA Marker-Assisted Selection and OPU-Derived IVF Embryo Transfer for the Production of High Quality Meat in Hanwoo I. Collection of Follicular Oocytes with Ultrasound-Guided Transvaginal Ovum Pick-Up from DNA Marker-proved Hanwo
본 연구는 고품질육의 DNA marker가 규떵된 한우로부터 초음파유노 난포란의 연속적 채취를 통하여 능력이 우수한 한우 수정란온 대량생산하는 방법의 확립과 이를 한우농가에 응용하고자 초음파 난자채취기를 이용하여 등지방층두께, 일당증체량, 근내지방도 및배최장근 단면적에 연관된 DNA marker를 보유하고 있는 한우 5두로부터 개체및 난포수, 채취방법, 회수한 난포란의 등급 등을 조사하였다. 한우 5두의 개체별 난포수는 6, 10, 5, 4 및 11회
This study was designed to establish the superior method for IVF embryos from DNA marker-proved Hanwoo cattle. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from unstimulated. immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local abesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analysed using the GLM procedure of SAS. Mean number of follicles identified on ultrasound was 5.5 2.9 in right and 4.3 2.8 in left ovaries, respectively. The highest follicles(16.62.6) were found in 5101 cow compared to others. Recovery rate of follicular oocytes in individual cow was highest in 5101 cow with 89.3% in > 2mm and 94.0% in 2mm follicles. Total recovery rate was significantly(P<0.01) higher in 2mm(85.7%, 130/154) than > 2mm follicles(74.2%, 201/271). Significantly more oocytcs of Grade IV were recovered from > 2mm follicles. Mean number follicles recovered was 4.83.7. 3.03.4 and 0.30.6 in 2mm, 2~6mm and 6mm follicles. respectively. Our results imply that the more fertilizablc oocytes can be recovered from invisible-immature follicles by the combination of simultaneous rectal ovarian-palpation and ultrasound-guided approach in Hanwoo cattle.