Hunter D D, Llinas R, Ard M, Merlie J P, Sanes J R
Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110.
J Comp Neurol. 1992 Sep 8;323(2):238-51. doi: 10.1002/cne.903230208.
The extracellular matrix component, s-laminin, is a homologue of the B1 subunit of laminin. S-laminin is concentrated in the synaptic cleft at the neuromuscular junction and contains a site that is adhesive for motor neurons, suggesting that it may influence neuromuscular development. To ascertain whether s-laminin may also play roles in the genesis of the central nervous system, we have examined its expression in the brain and spinal cord of embryonic and postnatal rats. S-laminin was not detectable in synapse-rich areas of adults. However, s-laminin was present in discrete subsets of three laminin-containing structures: (1) In the developing cerebral cortex, laminin and s-laminin were expressed in the subplate, a transient layer through which neuroblasts migrate and cortical afferents grow. Both laminin and s-laminin disappeared as embryogenesis proceeded; however, laminin was more widely distributed and present longer than s-laminin. (2) In the developing spinal cord, laminin was present throughout the pia. In contrast, s-laminin was concentrated in the pia that overlies the floor plate, a region in which extracellular cues have been postulated to guide growing axons. (3) In central capillaries, s-laminin appeared perinatally, an interval during which the blood-brain barrier matures. In contrast, laminin was present in capillary walls of both embryos and adults. To extend our immunohistochemical results, we used biochemical methods to characterize s-laminin in brain. We found that authentic s-laminin mRNA is present in the embryonic brain, but that brain-derived s-laminin differs (perhaps by a posttranslational modification) from that derived from nonneural tissues. We also used tissue culture methods to show that glia are capable of synthesizing "brain-like" s-laminin, and of assembling it into an extracellular matrix. Thus, glia may be one cellular source of s-laminin in brain. Together, these results demonstrate that s-laminin is present in the developing central nervous system, and raise the possibility that this molecule may influence developmental processes.
细胞外基质成分s-层粘连蛋白是层粘连蛋白β1亚基的同源物。s-层粘连蛋白集中于神经肌肉接头处的突触间隙,含有一个对运动神经元具有黏附性的位点,这表明它可能影响神经肌肉发育。为了确定s-层粘连蛋白是否也在中枢神经系统的发生过程中发挥作用,我们检测了其在胚胎期和出生后大鼠脑和脊髓中的表达情况。在成年大鼠富含突触的区域未检测到s-层粘连蛋白。然而,s-层粘连蛋白存在于三种含层粘连蛋白结构的离散亚群中:(1)在发育中的大脑皮质,层粘连蛋白和s-层粘连蛋白在中间层表达,中间层是神经母细胞迁移和皮质传入纤维生长通过的一个过渡层。随着胚胎发育的进行,层粘连蛋白和s-层粘连蛋白均消失;然而,层粘连蛋白分布更广泛,存在时间比s-层粘连蛋白更长。(2)在发育中的脊髓,层粘连蛋白存在于整个软脑膜。相比之下,s-层粘连蛋白集中在覆盖底板的软脑膜中,底板区域被认为存在细胞外信号引导生长中的轴突。(3)在中枢毛细血管中,s-层粘连蛋白在围产期出现,这是血脑屏障成熟的时期。相比之下,层粘连蛋白存在于胚胎和成年大鼠的毛细血管壁中。为了扩展我们的免疫组织化学结果,我们使用生化方法对脑中的s-层粘连蛋白进行表征。我们发现胚胎脑中存在真实的s-层粘连蛋白mRNA,但脑源性s-层粘连蛋白与非神经组织来源的s-层粘连蛋白不同(可能是通过翻译后修饰)。我们还使用组织培养方法表明神经胶质细胞能够合成“脑样”s-层粘连蛋白,并将其组装成细胞外基质。因此,神经胶质细胞可能是脑中s-层粘连蛋白的一个细胞来源。总之,这些结果表明s-层粘连蛋白存在于发育中的中枢神经系统中,并增加了该分子可能影响发育过程的可能性。