Oettinger H F, Thal G, Sasse J, Holtzer H, Pacifici M
Dev Biol. 1985 May;109(1):63-71. doi: 10.1016/0012-1606(85)90346-x.
Transverse frozen sections from the postcephalic region of stage 9-16 chick embryos and from the wing bud region of stage 17-31 embryos were stained with antibodies to the major extracellular matrix components of cartilage. These probes included unfractionated A1 and A2 antisera to the major cartilage proteoglycan, affinity-purified purified antibodies to the proteoglycan core protein and to Type II collagen, and a monoclonal antibody to keratan sulfate. In embryos as early as stage 10, notochord stained specifically with the keratan sulfate monoclonal antibody. At this stage the notochord, as well as surrounding tissues, were negative to cartilage proteoglycan and collagen antibodies. Positive staining with the latter probes was coordinately acquired by notochord cells and their accompanying sheath around stage 15, while surrounding tissues remained negative. At this stage, the ventral region of the perispinal cord sheath exhibited light staining with the proteoglycan and keratan sulfate antibodies though failing to react to Type II collagen antibodies. Positive staining of notochord and ventral spinal cord persisted through later developmental stages. As revealed by immunofluorescence, definitive vertebral chondroblasts first emerged at approximately stage 23 and definitive limb chondroblasts at stage 25. The results are discussed in terms of the possible multiple roles of notochord in early embryogenesis.
取自9至16期鸡胚头部后区域的横向冰冻切片以及17至31期胚胎翅芽区域的横向冰冻切片，用针对软骨主要细胞外基质成分的抗体进行染色。这些探针包括针对主要软骨蛋白聚糖的未分级A1和A2抗血清、针对蛋白聚糖核心蛋白和II型胶原的亲和纯化抗体，以及一种针对硫酸角质素的单克隆抗体。在早在10期的胚胎中，脊索用硫酸角质素单克隆抗体进行特异性染色。在这个阶段，脊索以及周围组织对软骨蛋白聚糖和胶原抗体呈阴性。在15期左右，脊索细胞及其伴随的鞘开始对后一种探针呈阳性染色，而周围组织仍为阴性。在这个阶段，脊髓鞘的腹侧区域用蛋白聚糖和硫酸角质素抗体染色较浅，但对II型胶原抗体无反应。脊索和脊髓腹侧的阳性染色在随后的发育阶段持续存在。免疫荧光显示，确定的椎体软骨细胞最早在大约23期出现，确定的肢体软骨细胞在25期出现。根据脊索在早期胚胎发生中可能的多种作用对结果进行了讨论。
