Patek C E, Vornhagen R, Rink H, Clayton R M
Exp Eye Res. 1986 Jul;43(1):29-40. doi: 10.1016/s0014-4835(86)80043-4.
We have compared the long-term developmental changes in water-insoluble protein expression by chick lens cells in vitro and in vivo. Crude membrane fractions were prepared by alkali treatment of the urea-insoluble protein fraction, and the proteins analysed by sodium dodecyl sulphate-polyacrylamide (SDS-PAGE) gel electrophoresis. The major component present in the urea-insoluble fraction of chick lens fibres, a 25,000 MW polypeptide (MIP-25K) was more abundant in adult (8 weeks) than day-old post-hatch chick lens fibre masses. MIP-25K was detected in differentiated but not predifferentiated lens cell cultures, and indirect immunofluorescence using anti-bovine MIP antiserum indicated that MIP-25K was localized in the lentoid bodies. Our findings indicate that the urea-insoluble protein profiles of long-term well-differentiated chick lens cell cultures are qualitatively very similar to the profiles of the lens fibres. The data also confirm that the expression of MIP-25K, rather than the expression of water-soluble crystallin protein, is a marker for lens cell differentiation, and confirm earlier reports, which have been disputed, that delta-crystallin (but not alpha-or beta-crystallin) is specifically associated with chick lens fibre membranes.
我们比较了鸡晶状体细胞在体外和体内水不溶性蛋白质表达的长期发育变化。通过对尿素不溶性蛋白质部分进行碱处理制备粗膜部分，并通过十二烷基硫酸钠-聚丙烯酰胺（SDS-PAGE）凝胶电泳分析蛋白质。鸡晶状体纤维的尿素不溶性部分中存在的主要成分，一种25,000分子量的多肽（MIP-25K），在成年（8周）鸡晶状体纤维团中比孵化后一天的鸡晶状体纤维团中更为丰富。在分化的而非预分化的晶状体细胞培养物中检测到MIP-25K，并且使用抗牛MIP抗血清的间接免疫荧光表明MIP-25K定位于类晶状体小体中。我们的研究结果表明，长期充分分化的鸡晶状体细胞培养物的尿素不溶性蛋白质谱在质量上与晶状体纤维的谱非常相似。数据还证实，MIP-25K的表达而非水溶性晶状体蛋白的表达是晶状体细胞分化的标志物，并证实了早期有争议的报道，即δ-晶状体蛋白（而非α-或β-晶状体蛋白）与鸡晶状体纤维膜特异性相关。
