Hadsell D L, Greenberg N M, Fligger J M, Baumrucker C R, Rosen J M
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Endocrinology. 1996 Jan;137(1):321-30. doi: 10.1210/endo.137.1.8536631.
To test the hypothesis that insulin-like growth factor I (IGF-I) regulates mammary gland development and lactation, the expression of both human (h) IGF-I and des(1-3)hIGF-I was targeted to the mammary gland in transgenic mice using a novel exon replacement strategy and the rat whey acidic protein (rWAP) gene regulatory sequences. Both transgenes expressed a 0.7-kilobase messenger RNA (mRNA). The abundance of WAP-IGE-I and WAP-DES mRNA on day 10 of lactation ranged from 0.2-1.0% and 0.2-13% of the endogenous mouse WAP mRNA, respectively. For WAP-DES mice, transgene expression was greatest from midpregnancy throughout lactation. Western blot analysis showed the presence of correctly processed hIGF-I in milk from these transgenic mice. This hIGF-I was capable of stimulating protein synthesis in cultured rat L6 myoblasts. Ligand blotting indicated changes in mammary gland secretion of IGFBP in response to WAP-DES expression. Histological analysis of mammary tissue from mice overexpressing des(1-3)hIGF-I showed incomplete mammary involution, ductile hypertrophy, and loss of secretory lobules associated with increased deposition of collagen. These changes are believed to occur through autocrine and paracrine effects of des(1-3)-hIGF-I on both epithelial and stromal cells.
为了验证胰岛素样生长因子I(IGF-I)调节乳腺发育和泌乳的假说,利用一种新的外显子替换策略和大鼠乳清酸性蛋白(rWAP)基因调控序列,将人(h)IGF-I和缺失(1-3)hIGF-I的表达靶向转基因小鼠的乳腺。两种转基因均表达0.7千碱基的信使核糖核酸(mRNA)。泌乳第10天时,WAP-IGF-I和WAP-DES mRNA的丰度分别为内源性小鼠WAP mRNA的0.2-1.0%和0.2-13%。对于WAP-DES小鼠,从妊娠中期到整个泌乳期转基因表达最高。蛋白质印迹分析表明这些转基因小鼠的乳汁中存在正确加工的hIGF-I。这种hIGF-I能够刺激培养的大鼠L6成肌细胞中的蛋白质合成。配体印迹表明,乳腺中IGFBP的分泌因WAP-DES的表达而发生变化。对过表达缺失(1-3)hIGF-I的小鼠乳腺组织进行组织学分析,结果显示乳腺 involution不完全、导管肥大以及分泌小叶丢失,同时伴有胶原蛋白沉积增加。据信这些变化是通过缺失(1-3)-hIGF-I对上皮细胞和基质细胞的自分泌和旁分泌作用而发生的。