Biophysical Sciences Institute, School of Biological and Biomedical Sciences, Durham University, South Road, Durham DH1 3LE, United Kingdom.
NETPark Incubator, Thomas Wright Way, Sedgefield, Durham TS21 3FD, United Kingdom.
Exp Eye Res. 2014 Mar;120(100):10-4. doi: 10.1016/j.exer.2013.12.004. Epub 2013 Dec 14.
Fibroblast growth factors play a key role in regulating lens epithelial cell proliferation and differentiation via an anteroposterior gradient that exists between the aqueous and vitreous humours. FGF-2 is the most important for lens epithelial cell proliferation and differentiation. It has been proposed that the presentation of FGF-2 to the lens epithelial cells involves the lens capsule as a source of matrix-bound FGF-2. Here we used immunogold labelling to measure the matrix-bound FGF-2 gradient on the inner surface of the lens capsule in flat-mounted preparations to visualize the FGF-2 available to lens epithelial cells. We also correlated FGF-2 levels with levels of its matrix-binding partner perlecan, a heparan sulphate proteoglycan (HSPG) and found the levels of both to be highest at the lens equator. These also coincided with increased levels of phosphorylated extracellular signal-regulated kinase 1 and 2 (pERK1/2) in lens epithelial cells that localised to condensed chromosomes of epithelial cells that were Ki-67 positive. The gradient of matrix-bound FGF-2 (anterior pole: 3.7 ± 1.3 particles/μm2; equator: 8.2 ± 1.9 particles/μm2; posterior pole: 4 ± 0.9 particles/μm2) and perlecan (anterior pole: 2.1 ± 0.4 particles/μm2; equator: 5 ± 2 particles/μm2; posterior pole: 1.9 ± 0.7 particles/μm2) available at the inner lens capsule surface was measured for the bovine lens. These data support the anteroposterior gradient hypothesis and provide the first measurement of the gradient for an important morphogen and its HSPG partner, perlecan, at the epithelial cell-lens capsule interface.
成纤维细胞生长因子在调节晶状体上皮细胞增殖和分化方面发挥着关键作用,其通过存在于房水和玻璃体之间的前后梯度来实现。FGF-2 对晶状体上皮细胞的增殖和分化最为重要。有人提出,FGF-2 向晶状体上皮细胞的呈现涉及晶状体囊作为基质结合的 FGF-2 的来源。在这里,我们使用免疫金标记法来测量平面培养物中晶状体囊内表面上的基质结合的 FGF-2 梯度,以观察可用于晶状体上皮细胞的 FGF-2。我们还将 FGF-2 水平与它的基质结合伴侣硫酸乙酰肝素蛋白聚糖 (HSPG) 多配体聚糖的水平相关联,发现两者的水平在晶状体赤道处最高。这也与在 Ki-67 阳性的上皮细胞中定位到凝聚的上皮细胞染色体的晶状体上皮细胞中的磷酸化细胞外信号调节激酶 1 和 2 (pERK1/2) 的水平增加相吻合。基质结合的 FGF-2(前极:3.7 ± 1.3 个/μm2;赤道:8.2 ± 1.9 个/μm2;后极:4 ± 0.9 个/μm2)和多配体聚糖(前极:2.1 ± 0.4 个/μm2;赤道:5 ± 2 个/μm2;后极:1.9 ± 0.7 个/μm2)的梯度可在牛晶状体的内晶状体囊表面上进行测量。这些数据支持前后梯度假说,并为重要形态发生因子及其 HSPG 伴侣多配体聚糖在上皮细胞-晶状体囊界面的梯度提供了首次测量。