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小鼠眼部组织中碱性成纤维细胞生长因子结合位点的个体发生

Ontogeny of basic fibroblast growth factor binding sites in mouse ocular tissues.

作者信息

Fayein N A, Courtois Y, Jeanny J C

机构信息

Unité de Recherches Gérontologiques, INSERM U. 118, Développement et Sénescence Cellulaire, Paris, France.

出版信息

Exp Cell Res. 1990 May;188(1):75-88. doi: 10.1016/0014-4827(90)90280-n.

Abstract

Basic fibroblast growth factor (bFGF) binding to ocular tissues has been studied by autoradiographical and biochemical approaches directly performed on sections during mouse embryonic and postnatal development. Frozen sections of embryos (9 to 18 days), newborns, and adults (1 day to 6 months) were incubated with iodinated bFGF. One specific FGF binding site (KD = 2.5 nM) is colocalized with heparan sulfate proteoglycans of the basement membranes and is heparitinase sensitive. It first appears at Day 9 around the neural tube, the optic vesicles, and below the head ectoderm and by Day 14 of embryonic development is found in all basement membranes of the eye. At Day 16, very intensely labeled patches appear, corresponding to mast cells which have been characterized by metachromatic staining of their heparin-rich granulations with toluidine blue. In addition to the latter binding, we have also observed a general diffuse distribution of silver grains on all tissues and preferentially in the ecto- and neuroectodermic tissues. From Days 17-18, there is heterogeneous labeling inside the retina, localized in the pigmented epithelium and in three different layers colocalized with the inner and outer plexiform layers and with the inner segments of the photoreceptors. This binding is heparitinase resistant but N-glycanase sensitive and may represent a second specific binding site corresponding to cellular FGF receptors (KD = 280 pM). Both types of binding patterns observed suggest a significant role for bFGF in eye development and physiology.

摘要

通过在小鼠胚胎期和出生后发育阶段直接对切片进行放射自显影和生化方法,研究了碱性成纤维细胞生长因子(bFGF)与眼部组织的结合情况。将胚胎(9至18天)、新生小鼠和成年小鼠(1天至6个月)的冰冻切片与碘化bFGF一起孵育。一个特异性FGF结合位点(KD = 2.5 nM)与基底膜的硫酸乙酰肝素蛋白聚糖共定位,且对肝素酶敏感。它最早在第9天出现在神经管、视泡周围以及头部外胚层下方,到胚胎发育第14天时,在眼睛的所有基底膜中都能发现。在第16天,出现了标记非常强烈的斑块,对应肥大细胞,通过用甲苯胺蓝对其富含肝素的颗粒进行异染性染色得以鉴定。除了后一种结合外,我们还观察到银颗粒在所有组织上普遍呈弥漫性分布,在外胚层和神经外胚层组织中分布更明显。从第17 - 18天开始,视网膜内部存在异质性标记,定位于色素上皮以及与内、外网状层和光感受器内段共定位的三个不同层。这种结合对肝素酶有抗性,但对N - 聚糖酶敏感,可能代表对应细胞FGF受体的第二个特异性结合位点(KD = 280 pM)。观察到的两种结合模式都表明bFGF在眼睛发育和生理过程中发挥着重要作用。

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