Guimarães A, Zaremba S, Hockfield S
Section of Neuroanatomy, Yale University School of Medicine, New Haven, Connecticut 06510.
J Neurosci. 1990 Sep;10(9):3014-24. doi: 10.1523/JNEUROSCI.10-09-03014.1990.
Monoclonal antibody Cat-301 recognizes a surface-associated proteoglycan on subsets of neurons in the mammalian CNS (Hockfield and McKay, 1983). The expression of Cat-301 immunoreactivity on Y cells in the cat LGN is sharply reduced by early visual deprivation (Sur et al., 1988). We employed an immunosuppression strategy (Hockfield, 1987) to further study alterations in the expression of experience-dependent molecules. Newborn BALB/c mice were injected with LGN from dark-reared cats to induce a suppression of the immune response to antigens expressed in visually deprived animals. These mice were then immunized with LGN from normal cats to elicit an immune response to antigens with an expression dependent on normal early visual experience. This strategy permitted the generation of monoclonal antibody Cat-304, which recognizes a surface-associated antigen on neuronal cell bodies and proximal dendrites, and which appears histologically identical to Cat-301. Further analyses show that Cat-304 and Cat-301 recognize different epitopes on the same 680-kDa chondroitin sulfate proteoglycan. We examined the effects of early visual deprivation on Cat-304 immunoreactivity in the LGN and visual cortex of cats. In LGN from normal cats, Cat-304 labels neurons in layers A, A1, and C, in interlaminar zones, and in the medial interlaminar nucleus. In LGN from dark-reared cats, the number of antibody-positive neurons is markedly reduced, and the cross-sectional area of the remaining positive neurons is smaller than normal. In cortical area 17 of normally reared cats, Cat 304-positive neurons are densely distributed in 2 bands, in layers IV and V/VI. Labeled neurons are also present in layers II and III. In area 17 of dark-reared cats, the number of antibody-positive neurons is reduced. The reduction in the number of labeled neurons is most pronounced in layers II/III and V/VI. Antibody-positive neurons are smaller in all cortical layers of dark-reared cats. The changes in the expression of Cat-301 immunoreactivity in dark-reared visual cortex and LGN are identical to those of Cat-304. The laminar differences in the effect of dark rearing on Cat-301 and Cat-304 expression in the visual cortex provides support for the suggestion that layer IV of cortical area 17 may be less susceptible to prolongation of plasticity by dark rearing than layers II/III and V/VI. Further, the biochemical and histological studies reported provide evidence that early visual experience regulates protein expression in the cat LGN and visual cortex.
单克隆抗体Cat-301可识别哺乳动物中枢神经系统中神经元亚群上的一种表面相关蛋白聚糖(霍克菲尔德和麦凯,1983年)。早期视觉剥夺会使猫外侧膝状体(LGN)中Y细胞上Cat-301免疫反应性的表达急剧降低(苏尔等人,1988年)。我们采用了一种免疫抑制策略(霍克菲尔德,1987年)来进一步研究经验依赖性分子表达的变化。给新生的BALB/c小鼠注射来自暗饲养猫的LGN,以抑制对视觉剥夺动物中表达的抗原的免疫反应。然后用来自正常猫的LGN对这些小鼠进行免疫,以引发对依赖正常早期视觉经验表达的抗原的免疫反应。这种策略产生了单克隆抗体Cat-304,它可识别神经元细胞体和近端树突上的一种表面相关抗原,并且在组织学上与Cat-301相同。进一步分析表明,Cat-304和Cat-301识别同一680 kDa硫酸软骨素蛋白聚糖上的不同表位。我们研究了早期视觉剥夺对猫LGN和视觉皮层中Cat-304免疫反应性的影响。在正常猫的LGN中,Cat-304标记A层、A1层和C层、层间区以及内侧层间核中的神经元。在暗饲养猫的LGN中,抗体阳性神经元的数量明显减少,其余阳性神经元的横截面积也比正常情况小。在正常饲养猫的17区皮层中,Cat 304阳性神经元密集分布在第IV层和第V/VI层的两条带中。标记的神经元也存在于第II层和第III层。在暗饲养猫的17区中,抗体阳性神经元的数量减少。标记神经元数量的减少在第II/III层和第V/VI层最为明显。暗饲养猫所有皮层层中的抗体阳性神经元都较小。暗饲养视觉皮层和LGN中Cat-301免疫反应性表达的变化与Cat-304的变化相同。暗饲养对视觉皮层中Cat-301和Cat-304表达影响的层间差异支持了这样一种观点,即与第II/III层和第V/VI层相比,17区皮层的第IV层可能对暗饲养延长可塑性的影响不太敏感。此外,所报道的生化和组织学研究提供了证据,表明早期视觉经验调节猫LGN和视觉皮层中的蛋白质表达。