首都医科大学学报
首都醫科大學學報
수도의과대학학보
JOURNAL OF CAPITAL UNIVERSITY OF MEDICAL SCIENCES
2015年
2期
262-269
,共8页
勾荣彬%江小华%巨荣凯%牛含虚%段德义%徐群渊
勾榮彬%江小華%巨榮凱%牛含虛%段德義%徐群淵
구영빈%강소화%거영개%우함허%단덕의%서군연
胰岛素样生长因子结合蛋白-4%转基因小鼠%神经发育
胰島素樣生長因子結閤蛋白-4%轉基因小鼠%神經髮育
이도소양생장인자결합단백-4%전기인소서%신경발육
insulin-like growth factor binding protein-4(IGFBP-4)%transgenic mice%neurodevelopment
目的：构建脑内过表达胰岛素样生长因子结合蛋白-4(insulin-like growth factor binding protein-4,IGFBP-4)的转基因小鼠,以研究IGFBP-4对脑发育的作用。方法构建神经元特异性启动子血小板源性生长因子β链( platelet derived growth factor βchain,PDGF-β)驱动的IGFBP-4表达载体,通过显微注射构建转基因小鼠。用PCR法检测小鼠子代基因组IGFBP-4 cDNA的整合情况,Western blotting法和免疫组织化学法观察IGFBP-4在脑内的表达丰度和分布,比较野生型和转基因小鼠的体质量和脑质量的改变。结果外源IGFBP-4 cDNA稳定整合于转基因小鼠基因组,IGFBP-4蛋白在成年小鼠脑内表达水平升高了267%,脑区IGFBP-4蛋白分布符合PDGF-β启动子驱动的蛋白表达特征,与野生型小鼠IGFBP-4部位相似。转基因小鼠体质量未见明显改变,整脑质量增加了8.4%(P<0.05),其中脑干增加了16.5%(P<0.05),其余脑区改变不明显。结论外源IGFBP-4转基因在小鼠基因组中得到整合并能稳定遗传和表达,并影响了部分脑区的发育,为脑发育研究提供了有价值的动物模型。
目的：構建腦內過錶達胰島素樣生長因子結閤蛋白-4(insulin-like growth factor binding protein-4,IGFBP-4)的轉基因小鼠,以研究IGFBP-4對腦髮育的作用。方法構建神經元特異性啟動子血小闆源性生長因子β鏈( platelet derived growth factor βchain,PDGF-β)驅動的IGFBP-4錶達載體,通過顯微註射構建轉基因小鼠。用PCR法檢測小鼠子代基因組IGFBP-4 cDNA的整閤情況,Western blotting法和免疫組織化學法觀察IGFBP-4在腦內的錶達豐度和分佈,比較野生型和轉基因小鼠的體質量和腦質量的改變。結果外源IGFBP-4 cDNA穩定整閤于轉基因小鼠基因組,IGFBP-4蛋白在成年小鼠腦內錶達水平升高瞭267%,腦區IGFBP-4蛋白分佈符閤PDGF-β啟動子驅動的蛋白錶達特徵,與野生型小鼠IGFBP-4部位相似。轉基因小鼠體質量未見明顯改變,整腦質量增加瞭8.4%(P<0.05),其中腦榦增加瞭16.5%(P<0.05),其餘腦區改變不明顯。結論外源IGFBP-4轉基因在小鼠基因組中得到整閤併能穩定遺傳和錶達,併影響瞭部分腦區的髮育,為腦髮育研究提供瞭有價值的動物模型。
목적：구건뇌내과표체이도소양생장인자결합단백-4(insulin-like growth factor binding protein-4,IGFBP-4)적전기인소서,이연구IGFBP-4대뇌발육적작용。방법구건신경원특이성계동자혈소판원성생장인자β련( platelet derived growth factor βchain,PDGF-β)구동적IGFBP-4표체재체,통과현미주사구건전기인소서。용PCR법검측소서자대기인조IGFBP-4 cDNA적정합정황,Western blotting법화면역조직화학법관찰IGFBP-4재뇌내적표체봉도화분포,비교야생형화전기인소서적체질량화뇌질량적개변。결과외원IGFBP-4 cDNA은정정합우전기인소서기인조,IGFBP-4단백재성년소서뇌내표체수평승고료267%,뇌구IGFBP-4단백분포부합PDGF-β계동자구동적단백표체특정,여야생형소서IGFBP-4부위상사。전기인소서체질량미견명현개변,정뇌질량증가료8.4%(P<0.05),기중뇌간증가료16.5%(P<0.05),기여뇌구개변불명현。결론외원IGFBP-4전기인재소서기인조중득도정합병능은정유전화표체,병영향료부분뇌구적발육,위뇌발육연구제공료유개치적동물모형。
Objective To establish a transgenic mouse which may over express insulin-like growth factor binding protein-4(IGFBP-4) in the brain, in order to explore the effect of IGFBP-4 on brain development. Methods The neuron-specific transgene expression cassette of IGFBP-4 was constructed and then microinjected into the fertilized mouse egg. The genotype of the transgenic mice was identified using PCR. The expression of IGFBP-4 protein in the brain was detected by Western blotting, and the spatial expression of IGFBP-4 was detected by immunohistochemistry. Both weights of body and brain of mice were measured. Results Exogenous IGFBP-4 cDNA was successfully integrated into the mouse genome and stably passaged. The expression of IGFBP-4 protein in the adult brain was increased by 267% in the transgenic mouse, while its spatial expression pattern was similar to that in the wild type mouse. The body weight was insignificantly changeable, but the whole brain weight was found an 8. 4% increase in transgenic mice(P<0. 05). Especially, the weight of brain stem increased by 16. 5%(P<0. 05), with no significant changes in other areas of the brain. Conclusion The exogenous IGFBP-4 cDNA was stably integrated into the transgenic mouse genome, and the over-expression of IGFBP-4 could affect development of certain part of the brain. The newly established IGFBP-4 transgenic mouse may therefore provide a useful animal model for investigating brain development.
