한국야생유래 행동이상 Mouse 에 있어서 체외수정에 의한 번식장애 개선과 정자의 동결보존

Improvement of Reproductive Disturbances by In Vitro Fertilization and Spermatozoa Cryopreservation in a Mouse Strain Showing Behavior Abnormality Derived from Korean Wild Mouse (Mus musculus molossinus)

본 연구는 한국야생 유래 행동이상 마우스(M. m. molossinus-tt@Kist)의 혈액학 및 혈액생화학치와, 체외수정란과 정자동결보존법, 체외수정과 수정란 이식기법을 이용한 번식장애 개선과 병원성 미생물이 제거된 무병마우스 생산을 위하여 실시하였다. 1. 5주령의 혈액학치에서 RBC, platelet치는 근교계에 비해 높게 나타났다. 혈액생화학치에서는 total cholesterol치가 근교계에 비해 높게 나타났으나 triglyceride, total protein, albumin치는 유사하였다. 2. 과배란 유기시의 평균 채란 수는 PMSG/hCG를 2.5/2.5 lU 투여군에서 11.6개, 5.0/5.0 IU 투여군에서 12.7개로 통계학적 유의 차는 인정되지 않았다. 3. 2.5/2.5 lU 투여군과 5.0/5.0 lU 투여군의 수정율은 각각 87.9%와 52%로 2.5/2.5 lU 부여군이 유의성있게 높은 성적을 나타내었고(p<0.05), 2세포기로의 발달율도 각각 유의성이 인정되는 99.0%와 90.6%였다 (p<0.05). 4. 동결정자를 이용한 체외수정에서의 수정율은 24.8%로 신선정자를 사용했을 때의 87.9% 보다 낮은 성적이었다. 5. 체외 수정란의 이식후의 산자율은 동결 2세포기 체외수정란의 경우 5마리(6.6%), 동결 정사를 이용한 체외 수정란의 경우 6마리(19.4%)와 대조군의 체외수정란의 경우 10마리(21.7%)의 새끼를 얻었다. 6. 이식후 출산한 산자의 미생물학적 검사에서 MHV(Mouse hepatitis virus)와 Staphylo-coccus aureus 등의 병원성 미생물이 무병대리모를 이용한 수정란이식에 의해 제거되었음을 확인하였다.

The present study was performed to improve the reproductive disturbance as well as the elimination of microbiological contamination for animals bred under conventional conditions followed by in vitro fertilization and embryo transfer techniques including embryo and sperm freezing, using a mouse strain(M. m. molossinus-tt＠Kist) showing the abnormal behavior disorder derived from Korean wild mice (Mus musculus molossinus). Moreover, hematological and serum biochemical analyses were also carried out to obtain the basic data of this mouse strain The results are summarized as follows: 1. In comparison with hematological data, the numbers of RBC and platelet of this mouse strain were appeared as the higher value those that of the same aged inbred strains such as BALB/c, DBA/2, C57BL/6 and C3H /Hen. However, no differences were found in values of WBC, Hb and Ht. Moreover, total cholesterol of this strain showed a low value but triglyceride, total protein and albumin values were similar as in inbred strains. 2. The average numbers of superovulated oocytes treated with 2.5/2.5 IU and 5.0/5.0 IU of PMSG/hCG were 11.6 and 12.7, respectively. The fertilization rates of 2.5/2.5 IU PMSG /hCG treatment(87.9%) was higher than 5.0/5.0 IU treatment(52.0%) (p＜0.05) and the developmental rate of 2 cell stage embryos were 외 so appeared as higher value 99.0% and 90.6%, respectively. 3. The rates of in vitro fertilization treated with frozen sperm(24.8%) was significantly lower than of that fresh sperm(87.9%), (p＜0.05). 4. The five, six and ten heads of offspring were obtained from frozen-thawed 2 cell embryos by in vitro fertilized, 2 cell embryos from in vitro fertilized by frozen-thawed spermatozoa. and 2 cell embryos by in vitro fertilization, respectively. These offspring developed the expected disease about 2 weeks after birth, which was confirmed that the disease character of this mutant mouse strain was reliably reproduced. 5. MHV(Mouse hepatitis virus) and Staphylococcus aureus were successfully eliminated from conventional animals by in vitro fertilization-embryo transfer and the use of SPF recipient animals.