Immunohistochemical analysis of proteoglycan biosynthesis during early development of the chicken cornea.
Immunohistochemical analysis of proteoglycan biosynthesis during early development of the chicken cornea.
复制标题
鸡角膜早期发育过程中蛋白多糖生物合成的免疫组织化学分析。
DOI：
10.1093/oxfordjournals.jbchem.a022520
复制
发表时间：
1999
影响因子：
2.7
通讯作者：
K. Nakazawa
中科院分区：
文献类型：
--
作者：
I. Takahashi;Y. Nakamura;Y. Hamada;K. Nakazawa
制备了鸡角膜硫酸角蛋白聚糖和硫酸软骨素蛋白聚糖核心蛋白抗体，并采用亲和柱纯化。利用这些抗体和硫酸角蛋白单克隆抗体5- d -4（市售），免疫组织化学方法测定了蛋白多糖在发育中的角膜（胚龄5 ~ 17天和孵化后2天）的定位。第5天角膜中未检测到硫酸角蛋白多糖抗原，但在第6天，即间充质细胞侵袭原代间质约12小时后，在整个间质中均检测到硫酸角蛋白多糖抗原。第5天角膜中抗原的缺失通过角膜提取物的Western blotting证实。免疫组化5-D-4抗体显示，第6 ~ 7天角膜的硫酸角蛋白链是欠硫酸化的，因为染色明显弱于第8天的角膜。此外，在孵化后的第7 - 17天和第2天，在整个基质中均检测到硫酸角蛋白多糖抗原，但在第13天及之后的上皮层中未检测到抗原：由于在显微镜下直到第13天才明显观察到上皮层，因此不知道第6 - 12天上皮是否合成了硫酸角蛋白多糖。第5天在角膜中检测到硫酸软骨素蛋白多糖抗原，并且在孵化后第6天至第2天在整个基质中均检测到硫酸软骨素蛋白多糖抗原。此外，在第13天及之后的上皮层中未检测到硫酸软骨素蛋白多糖。这些结果表明，在第5.5天至第6天，侵袭原代基质的基质细胞合成硫酸角蛋白多糖，而侵袭前上皮细胞和/或内皮细胞合成硫酸软骨素蛋白多糖，侵袭后基质细胞也合成硫酸软骨素蛋白多糖。
Antibodies to core proteins of chicken corneal keratan sulfate proteoglycan and chondroitin sulfate proteoglycan were prepared and purified by use of an affinity column. Using these antibodies and monoclonal antibody 5-D-4 to keratan sulfate (commercial), the localization of proteoglycans in developing corneas (Days 5 to 17 of embryonic age and 2 days after hatching) was determined immunohistochemically. Keratan sulfate proteoglycan antigen was not detected in cornea on Day 5, but it was detected uniformly over the whole stroma on Day 6, ca. 12 h after invasion of the primary stroma by mesenchymal cells. The absence of the antigen in cornea of Day 5 was confirmed by Western blotting of the corneal extract. Immunohistochemistry with 5-D-4 antibody revealed that the keratan sulfate chain was undersulfated in corneas of Days 6 to 7, because the staining was much weaker than that in cornea of Day 8. In addition, keratan sulfate proteoglycan antigen was detected uniformly over the whole stroma on Days 7 to 17 and 2 days after hatching, but not in the epithelial layer on Day 13 and after: because the epithelial layer was clearly not observed on photomicrographs until Day 13, it is not known whether keratan sulfate proteoglycan was synthesized by the epithelium during Days 6 to 12. In contrast, chondroitin sulfate proteoglycan antigen was detected in cornea on Day 5 and also, like keratan sulfate proteoglycan, uniformly over the whole stroma on Day 6 through 2 days after hatching. Furthermore, the chondroitin sulfate proteoglycan was not detected in the epithelial layer on Day 13 and after. These results show that keratan sulfate proteoglycan is synthesized by the stromal cells which invade the primary stroma between Day 5.5 and 6, while chondroitin sulfate proteoglycan is synthesized by epithelial and/or endothelial cells before the invasion, and also by the stromal cells after the invasion.
影响因子：
6.1
作者：
Cao JQ;Liang YY;Li YQ;Zhang HL;Zhu YL;Geng J;Yang LQ;Feng SW;Yang J;Kong J;Zhang C
通讯作者：
Zhang C
影响因子：
3.9
作者：
LI, W;VERGNES, JP;HASSELL, JR
通讯作者：
HASSELL, JR
DOI：
10.1016/s0021-9258(18)48500-6
发表时间：
1992-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
T. C. Blochberger;J. Vergnes;J. Hempel;J. Hassell
通讯作者：
T. C. Blochberger;J. Vergnes;J. Hempel;J. Hassell