The detection of a precartilage, blastema-specific marker.
The detection of a precartilage, blastema-specific marker.
复制标题
检测前软骨、胚基特异性标记物。
DOI：
10.1016/0012-1606(87)90240-5
复制
发表时间：
1987
影响因子：
2.7
通讯作者：
Solursh,M
中科院分区：
文献类型：
--
作者：
Aulthouse,AL;Solursh,M
在活体和高密度细胞培养中，间充质细胞聚集体被称为体内胚泡，先于软骨分化。半乳糖特异性凝集素，花生凝集素(PNA)，已被证明是胚芽特异性的(B.Zimmermann和M.Thies，1984，组织化学81,353-361)。PNA似乎是活体和体外软骨前细胞聚集物的标志。对24期雏鸡翼芽冰冻切片进行PNA-罗丹明双染和针对II型胶原抗体的间接免疫荧光染色。PNA染色较深，向远端延伸至II型胶原水平。24期雏鸡翅芽的高密度培养也评估了PNA结合的分布。培养16小时后，PNA结合出现的时间最早。PNA染色的区域与苏木精染色的细胞聚集体一致。PNA染色被50 MMD(+)-半乳糖抑制，对1%睾丸透明质酸酶不敏感。16小时后尚无阿尔西亚蓝染色结节出现。软骨前、胚泡特异性标记物的存在，以及在培养的软骨前聚集体中的存在，暗示了这两种情况下软骨形成的相似性。阶段19肢芽培养没有形成结节，但确实形成了PNA阳性的聚集体。此外，在胶原凝胶上或在细胞松弛素D处理后分化为软骨细胞的单个细胞缺乏PNA结合物质。这些结果表明这种材料是软骨前聚集体所特有的。PNA阳性物质呈胞外分布，用0.5Mguanine盐酸盐短暂提取后除去。
Mesenchymal cell aggregates, termed blastemain vivo, precede cartilage differentiationin vivoand in high-density cell cultures. The galactose specific lectin, peanut agglutinin (PNA), has been shown to be blastema specific (B. Zimmermann and M. Thies, 1984,Histochemistry81,353–361). PNA appears to be a marker for precartilage cellular aggregates bothin vivoandin vitro. Frozen sections of stage 24 chick wing buds were double stained with PNA-rhodamine and by indirect immunofluorescence with antibody directed against type II collagen. The PNA stained the humeral blastema intensely and extended distal to the level of type II collagen. High-density cultures of stage 24 chick wing buds were also evaluated for the distribution of PNA binding. Sixteen-hour cultures showed the earliest consistent appearance of PNA binding. The PNA-stained areas coincided with hematoxylin-stained cell aggregates. PNA staining was inhibited by 50 mMd(+)-galactose and was not sensitive to 1% testicular hyaluronidase pretreatment. No Alcian blue-staining nodules were present yet at 16 hr. The presence of a precartilage, blastema-specific markerin situ, as well as in precartilage aggregates in cultures, suggests the similarities in chondrogenesis between these two conditions. Stage 19 limb bud cultures did not form nodules but did form aggregates that were PNA positive. Furthermore, single cells that differentiated into chondrocytes on collagen gels or after cytochalasin D treatment lacked PNA-binding material. These results suggest that this material is specific to precartilage aggregates. The PNA-positive material was extracellular in distribution and was removed after brief extraction with 0.5Mguanidine hydrochloride.