Differential Expression of Micro-Heterogeneous LewisX-Type Glycans in the Stem Cell Compartment of the Developing Mouse Spinal Cord
Differential Expression of Micro-Heterogeneous LewisX-Type Glycans in the Stem Cell Compartment of the Developing Mouse Spinal Cord
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DOI：
10.1007/s11064-013-1048-6
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发表时间：
2013-06-01
影响因子：
4.4
通讯作者：
Faissner, Andreas
中科院分区：
文献类型：
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作者：
Karus, Michael;Hennen, Eva;Faissner, Andreas
复杂的聚糖结构和它们各自的载体分子通常以细胞类型特异性方式表达。因此，聚糖可用于富集特定细胞类型，如神经前体细胞（NPC）。我们最近已经表明，单克隆抗体487（LeX）和5750（LeX）差异检测发育中的小鼠前脑中NPC上的LewisX（LeX）聚糖。在这里，我们分析了两种抗体在胚胎小鼠脊髓发育后期的染色模式。在E13.5，两种抗体均强烈标记中央管区域。沿着这些路线，他们主要在该年龄的巢蛋白阳性NPC上检测LeX聚糖。此外，我们表明，脊髓NPC培养的自由浮动的神经球显示出高的免疫反应性，这两种抗体。在这种情况下，我们还证明了487（LeX）抗体可用于从混合的E13.5脊髓细胞悬液中消耗神经球形成NPC的亚群。接近胚胎发生结束时，对两种抗体的总体免疫反应性增加，染色似乎非常扩散。然而，5750（LeX）抗体仍然标记中央管区域。免疫反应性的增加与细胞外基质分子腱生蛋白C和受体蛋白酪氨酸磷酸酶β/ζ（两种潜在的LeX载体蛋白）的表达增加相关。与此一致，免疫沉淀分析证实了腱生蛋白C作为胚胎小鼠脊髓中的LeX载体蛋白。然而，在没有腱生蛋白C的情况下，对两种抗体的免疫反应性似乎仅受到轻微影响，从而反对腱生蛋白C是主要的LeX载体。总之，我们的研究提供了一些新的见解复杂的表达LeX聚糖和潜在的载体蛋白在小鼠脊髓的发展。
Complex glycan structures and their respective carrier molecules are often expressed in a cell type specific manner. Thus, glycans can be used for the enrichment of specific cell types such as neural precursor cells (NPCs). We have recently shown that the monoclonal antibodies 487(LeX) and 5750(LeX) differentially detect the LewisX (LeX) glycan on NPCs in the developing mouse forebrain. Here, we analysed the staining pattern of both antibodies during late embryonic mouse spinal cord development. At E13.5 both antibodies strongly label the central canal region. Along these lines they detect the LeX glycan primarily on Nestin-positive NPCs at that age. Moreover, we show that spinal cord NPCs cultured as free floating neurospheres display a high immunoreactivity to both antibodies. In that context, we also demonstrate that the 487(LeX) antibody can be used to deplete a subpopulation of neurosphere forming NPCs from a mixed E13.5 spinal cord cell suspension. Towards the end of embryogenesis the overall immunoreactivity to both antibodies increases and the staining appears very diffuse. However, the 5750(LeX) antibody still labels the central canal region. The increase in immunoreactivity correlates with an expression increase of the extracellular matrix molecules Tenascin C and Receptor Protein Tyrosine Phosphatase beta/zeta, two potential LeX carrier proteins. In line with this, immunoprecipitation analyses confirmed Tenascin C as a LeX carrier protein in the embryonic mouse spinal cord. However, the immunoreactivity to both antibodies appears only to be marginally affected in the absence of Tenascin C, arguing against Tenascin C being the major LeX carrier. In conclusion our study gives some novel insights into the complex expression of LeX glycans and potential carrier proteins during the development of the mouse spinal cord.