Schlosshauer B, Blum A S, Mendez-Otero R, Barnstable C J, Constantine-Paton M
Department of Biology, Yale University, New Haven, Connecticut 06511.
J Neurosci. 1988 Feb;8(2):580-92. doi: 10.1523/JNEUROSCI.08-02-00580.1988.
The JONES monoclonal antibody has been immunocytochemically associated with regions of the developing rat brain where cell and axon migrations are occurring (Mendez-Otero et al., 1986, 1988). In the present study the antigens recognized by the JONES antibody were analyzed in a variety of brain regions and at developmental ages selected to correspond to the preceding immunocytochemical observations. In accordance with earlier results from retina, JONES binding could not be detected in SDS gels from developing brain. Binding of the antibody was, however, prominent in chloroform/methanol extracts of the same tissues, and it was completely removed from tissue sections by brief chloroform/methanol form/methanol treatment. Enzymatic analyses of chloroform/methanol extracts indicated that the JONES epitope was sensitive to neuraminidase but insensitive to proteases. Overlay assays on developed high-performance thin-layer chromatographic plates (HPTLC) indicate that in all regions the JONES epitope resides on 2 or 3 ganglioside bands, depending on the age examined. These bands migrate between ganglioside standards GD 1a and GM2 on HPTLC plates and have been designated GJ1, GJ2, and GJ3, with the higher number designating the more rapidly migrating species. Occasionally, additional bands migrating in the range of polysialogangliosides were observed. The pattern of expression of GJ species was studied in forebrain, retina, and cerebellar tissue taken from embryonic day 18 (E18), postnatal day 0 (P0), P7, P14, and adult animals. Both region-specific differences in the relative prominence of each band and stage-specific differences in the total amount of the JONES gangliosides were detected. The stage-specific differences in the amount of JONES antigens are well correlated with the developmental periods of maximal cell migration in each region. While the JONES gangliosides are most prominent in forebrain before birth, in they are most prominent during the first 2 postnatal weeks. In cerebellum, JONES antigen expression is more pronounced during the 2 periods of cell migration in this tissue. In retina, the more rapidly migrating GJ3 band was the most prominent band at all stages examined, and this same band is retained in the adult. In cerebellum and forebrain GJ3 is also the most pronounced band during development. However, in contrast to the retina, the more slowly migrating GJ1 band is retained in adult forebrain and cerebellum. A variety of non-brain tissues have also been examined for the presence of the JONES antigens.(ABSTRACT TRUNCATED AT 400 WORDS)
琼斯单克隆抗体已通过免疫细胞化学方法与正在发生细胞和轴突迁移的发育中大鼠脑区相关联（门德斯 - 奥泰罗等人，1986年，1988年）。在本研究中，对琼斯抗体所识别的抗原在各种脑区以及选定的发育年龄进行了分析，这些年龄与之前的免疫细胞化学观察结果相对应。与视网膜早期结果一致，在发育中脑的十二烷基硫酸钠凝胶中未检测到琼斯结合。然而，该抗体在相同组织的氯仿/甲醇提取物中结合显著，并且通过短暂的氯仿/甲醇/甲醇处理可从组织切片中完全去除。氯仿/甲醇提取物的酶分析表明，琼斯表位对神经氨酸酶敏感，但对蛋白酶不敏感。在展开的高效薄层色谱板（HPTLC）上进行的覆盖分析表明，在所有区域中，根据所检查的年龄，琼斯表位位于2条或3条神经节苷脂带上。这些条带在HPTLC板上的神经节苷脂标准品GD1a和GM2之间迁移，已被命名为GJ1、GJ2和GJ3，数字越大表示迁移越快的种类。偶尔会观察到在多唾液酸神经节苷脂范围内迁移的其他条带。研究了取自胚胎第18天（E18）、出生后第0天（P0）、P7、P14和成年动物的前脑、视网膜和小脑组织中GJ种类的表达模式。检测到各条带相对突出程度的区域特异性差异以及琼斯神经节苷脂总量的阶段特异性差异。琼斯抗原量的阶段特异性差异与每个区域细胞迁移最大的发育时期密切相关。虽然琼斯神经节苷脂在出生前的前脑中最突出，但在出生后的前两周最突出。在小脑中，琼斯抗原表达在该组织细胞迁移的两个时期更为明显。在视网膜中，迁移较快的GJ3带在所有检查阶段都是最突出的条带，并且在成年期也保留该条带。在小脑中，GJ3在发育过程中也是最明显的条带。然而，与视网膜不同，迁移较慢的GJ1带保留在成年前脑和小脑中。还检查了多种非脑组织中是否存在琼斯抗原。（摘要截短于400字）
