표유동물의 신진대사와 세포성장, 증식 및 분화에 관여한다고 알려진 인슐린의 착상전 배아에 어떤 영향을 미치는지 알기위해, 시헌관내에서 생쥐 수정란을 포배시기까지 일정한 배양액 즉 인슐린과 2-cell 배아 발육정지 현상을 일으키는 hypoxanthine 그리고 EDTA를 각각 혹은 혼합투여한 배양액에서 배양하였다. 배아난할 및 배아발달을 비교 관찰하였고, 포배발달에 필요한 인슐린의 EC50을 알아보았으며, 아울러 면역세포화확적 방법으로 인슈린 수용체을 확인하여 이를 통해 작용하는 인슐린의 단백질 합성과의 관계를 배아발달 시기별로 측정하여 다음과 같은 결론을 얻었다. 1. mEBSS 배양액에서 생쥐 수정란의 1차 난할율은 첨가제와 관계없이 약 90%였으나 이후 상실배 및 포배발달은 대조군에서는 40.6%와 23.6%였고, hypoxanthine 투여군에서는 10%이하로 차단된 반 면(P<0.001), 인슐린과 EDTA투여군에서는 80%와 60%이상의 높은 배아발달을 보였다(P<0.001) 3. 시험관내에서 2-cell 배아로부터 포배발달을 유도할 수 있는 인슐린의 EC50는 13.1nM이었다. 4. 면역세포화학적 염색법으로 포배에서 인슐린수용체을 발현시켰다. 5. 인슐린 투여군 및 대조군 모두에서 배아발달에 따라 단백질 합성을 증가시켰지만 인슐린 투여군 이 대조군에 비해 8-cell시기에 13%(P<0.05), 상실배에서 64%(P<0.05), 포배에서 99%(P<0.01)더 높은 효과를 나타냈다. 6. 인슈린 항체투여는 배아 단백질 합성에는 효과가 없었으나 인슐린의 촉진작용을 상실배와 포배 시기게 효과적으로 억제하였다(P<0.05 와 P<0.01).이상과 같은 실험결과로 볼 때 인슐린은 착상전 생쥐 배아에서 배형성 및 발달에 중요한 성장인자로써 작용하고 있다고 사료된다, 위 논문은 가톨릭 중앙의료원 학술연구 조성비로 이우러짐.

Although insulin is recognized as an important fetal growth factor, interaction with embryonic cells during the early development stages prior to the appearance of a functional pancreas has not been definde. But there is evidence that insulin binding to the cells of the preimplantation mouse embryos can be detected first at the morula stage, while autoradiographic studies have confirmed that insulin binding is receptor mediated. Other sutides have provided evidence that insulin is capable of stimulating glucose transport and protein synthesis in preimplantation mouse embryos. In this study, to investigate the effect of insulin on development of mouse preimplantation embryos, we performed the culture of mouse embryos from 1-cell zygote to blastocyst in modified Earle`s blanced salf solution(mEBSS), A chemically defined simple medium, supplemented with porcine insulin,hypoxanthine and/or ethylenediaminetetraacetic acid(EDTA), and the embryo development was scored at 24hours(2-cell)and96hours(morula,blastocyst)of culture. We also measured EC for blastocyst development form 2-cell enbryos inmEBSS supplemented with various concentrations of insulin, And the mouse embryos were stained to express the insulin receptor by immunohistochemical nethod usingan anti-insulin receptor immunoglobulinG.In addition to the immunohistochemical studies for localization of insulim receptor,experiments were undertaken to evaluate whether of not insulin exerts a stimulation of protein synthesis at early stages of mammalian enbryo genesis. The results were as follows: 1. The first cleavage rate of embryos derived from random-bred female mice was appoximately 90% in all experimental groups. In addition, although 40.6%and23.6% of 2-cell embryos developed to morula and blastocyst stage, respectively,in the control group, fewer than10% developed beyond 2-cell stage the hypoxanthine-supplemented medium(p<0.001)but significantly more embryos developed to morula(>80%)and blastocysts(>60%)in the presence of insulin of EDTA (p<0.001). 2. Insulin caused an increase of 20% in blastocyst development in the presence of EDTA(p<0.01)and dffectively reversed the "2-cell block" by hypoxanthine(p<0.001). 3. The EC of insulin for induction of blastocyst development form 2-cell embryos was 13.1nM in the presence of0.45 bovine serum albumin. 4. Insulin receptor was expressed by immunohistochemical staining using an antiinsulin receptor immunoglobulin G at the blastocyst stage of mouse embryos. 5. Culture with insulin for 4hour caused an increase of13% in protein synthesis but it completely blocked the effect of insulin in both-supplemented medium at the mourla and blastocyst stage(P<0.05 and P<0.01, respectively). From the above results, we conclude that insulin may play a role as an important growth factor in embryogenesis early development of mouse preimplantation embryos by the insulin-receptor mediated interaction