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猪主动脉中胶原蛋白和弹性蛋白基因表达的纵向梯度

Longitudinal gradients of collagen and elastin gene expression in the porcine aorta.

作者信息

Davidson J M, Hill K E, Mason M L, Giro M G

出版信息

J Biol Chem. 1985 Feb 10;260(3):1901-8.

PMID:3838176
Abstract

The physical and chemical properties of the mammalian aorta are known to vary as a function of distance from the heart. These properties are highly dependent collagen and elastic fibers. In order to evaluate the mechanisms which regulate the accumulation of these two connective tissue proteins, gene expression was evaluated at both the biosynthetic and messenger RNA levels. Short-term (3 h) explant cultures of the medial portion of four segments of the descending aorta in newborn pigs were incubated in the presence of [3H] proline. Collagen production was quantified by collagenase digestion and elastin production was determined by immunoprecipitation. Between the conus arteriosus and the bifurcation of the iliac arteries, relative collagen synthesis increased 2-fold (from 5.8 to 12.0% of total protein synthesis), while relative elastin synthesis declined 10-fold (from 16.4 to 1.6% of total protein synthesis). Similarly, collagen production increased more than 7-fold (from 6.7 to 49.8 X 10(3) molecules/cell/h) while elastin production was reduced more than 3-fold (from 71.8 to 21.0 X 10(3) molecules/cell/h) along this developmental gradient. Elastin synthesis appeared to be controlled to a significant extent by the availability of elastin mRNA, since both cell-free translation and molecular hybridization to a cloned elastin gene probe showed gradients of elastin gene expression. Similarly, collagen synthesis was apparently regulated, at least in part, by an inverse gradient of collagen mRNA, as measured with a cloned cDNA for the pro-alpha 1(I) collagen gene. Marked changes in the amount of non-elastin protein synthesis accompanied differentiation and accounted for larger changes in relative synthesis. These results suggest that the phenotype of the cells of the porcine artery wall is distinct in different regions of this organ at this developmental stage.

摘要

哺乳动物主动脉的物理和化学性质已知会随着与心脏距离的变化而变化。这些性质高度依赖于胶原蛋白和弹性纤维。为了评估调节这两种结缔组织蛋白积累的机制,在生物合成和信使核糖核酸水平上对基因表达进行了评估。将新生猪降主动脉四个节段中间部分的短期(3小时)外植体培养物在[3H]脯氨酸存在下孵育。通过胶原酶消化定量胶原蛋白的产生,通过免疫沉淀测定弹性蛋白的产生。在动脉圆锥和髂动脉分叉之间,相对胶原蛋白合成增加了2倍(从总蛋白合成的5.8%增加到12.0%),而相对弹性蛋白合成下降了10倍(从总蛋白合成的16.4%下降到1.6%)。同样,沿着这个发育梯度,胶原蛋白的产生增加了7倍多(从6.7×10³分子/细胞/小时增加到49.8×10³分子/细胞/小时),而弹性蛋白的产生减少了3倍多(从71.8×10³分子/细胞/小时减少到21.0×10³分子/细胞/小时)。弹性蛋白的合成似乎在很大程度上受弹性蛋白信使核糖核酸可用性的控制,因为无细胞翻译和与克隆的弹性蛋白基因探针的分子杂交都显示出弹性蛋白基因表达的梯度。同样,胶原蛋白的合成显然至少部分地由胶原蛋白信使核糖核酸的反向梯度调节,这是用针对原α1(I)胶原蛋白基因的克隆互补脱氧核糖核酸测量的。非弹性蛋白合成量的显著变化伴随着分化,并导致相对合成的更大变化。这些结果表明,在这个发育阶段,猪动脉壁细胞的表型在该器官的不同区域是不同的。

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