Kind P, Blakemore C, Fryer H, Hockfield S
University Laboratory of Physiology, University of Oxford, United Kingdom.
Cereb Cortex. 1994 Jul-Aug;4(4):361-75. doi: 10.1093/cercor/4.4.361.
To identify proteins that play a role in the development of the mammalian visual cortex, we have used an immunosuppression and rapid immunization strategy to generate monoclonal antibodies to antigens that are present in area 17 of the cat during the peak of cortical plasticity but are downregulated near the end of the plastic period. We report here the immunohistochemical and immunobiochemical characterization of six monoclonal antibodies that identify antigens preferentially expressed in the cat visual cortex at 5 weeks of age. Monoclonal antibodies Cat-305 and Cat-306 detect three immunoreactive elements that are not present at birth but are present at 5 weeks. The majority of immunoreactivity is associated with a population of cells in the white matter that are absent at 15 weeks of age. At both 5 and 15 weeks, a very small number of neurons show intense immunoreactivity throughout all processes, resembling that achieved with a Golgi stain. In addition, a diffuse band of immunoreactivity in layer IV is largely restricted to cortical areas 17 and 18. Cat-307 recognizes a 150 kDa soluble protein present in small cytoplasmic inclusions. These cytoplasmic "dot" are present in all layers, but are most prominent in layer V. Cat-307 immunoreactivity is present at birth and is completely downregulated by 15 weeks. Cat-104 and Cat-105 recognize a 200 kDa insoluble protein present at birth and at 5 weeks, but markedly downregulated by 15 weeks. At birth, the white matter, subplate, and layer I are most densely labeled, while at 5 weeks labeling is densest in layers II, III, and V. Cat-402 recognizes a number of high-molecular-weight antigens that are differentially expressed at 5 and 15 weeks of age. Stained non-neuronal cells that resemble protoplasmic astrocytes are present in all layers at both 5 and 15 weeks. At 5 weeks, but not at birth or 15 weeks, darkly immunoreactive radial processes are observed that run through the full depth of the cortex. We show here that immunoreactivity for several different monoclonal antibodies is detected selectively during the period of maximal developmental plasticity. The results demonstrate that the cat visual cortex at 5 weeks of age is molecularly distinct from the cortex at 15 weeks.
为了鉴定在哺乳动物视皮层发育过程中发挥作用的蛋白质,我们采用了免疫抑制和快速免疫策略,以产生针对猫17区在皮层可塑性高峰期存在但在可塑性期结束时下调的抗原的单克隆抗体。我们在此报告六种单克隆抗体的免疫组织化学和免疫生物化学特征,这些抗体可识别在5周龄时在猫视皮层中优先表达的抗原。单克隆抗体Cat - 305和Cat - 306检测到三种出生时不存在但在5周时存在的免疫反应性元件。大部分免疫反应性与白质中的一群细胞相关,这群细胞在15周龄时不存在。在5周和15周时,都有极少数神经元在所有突起中显示出强烈的免疫反应性,类似于用高尔基染色法获得的结果。此外,IV层中的弥漫性免疫反应带主要局限于皮层17区和18区。Cat - 307识别存在于小细胞质内含物中的一种150 kDa可溶性蛋白质。这些细胞质“点”存在于所有层中,但在V层中最为明显。Cat - 307免疫反应性在出生时存在,并在15周时完全下调。Cat - 104和Cat - 105识别一种出生时和5周时存在但在15周时明显下调的200 kDa不溶性蛋白质。出生时,白质、亚板层和I层标记最密集,而在5周时,II层、III层和V层标记最密集。Cat - 402识别一些在5周和15周龄时差异表达的高分子量抗原。在5周和15周时,所有层中都存在类似于原浆性星形胶质细胞的染色非神经元细胞。在5周时,但在出生时或15周时未观察到,可见深色免疫反应性的放射状突起贯穿皮层的全层深度。我们在此表明,在最大发育可塑性期间选择性地检测到了几种不同单克隆抗体的免疫反应性。结果表明,5周龄时的猫视皮层在分子水平上与15周龄时的皮层不同。