Integrins Contribute to Initial Morphological Development and Process Outgrowth in Rat Adult Hippocampal Progenitor Cells
Integrins Contribute to Initial Morphological Development and Process Outgrowth in Rat Adult Hippocampal Progenitor Cells
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DOI：
10.1007/s12031-009-9211-x
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发表时间：
2010-03-01
影响因子：
3.1
通讯作者：
Sakaguchi, Donald S.
中科院分区：
文献类型：
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作者：
Harper, Matthew M.;Ye, Eun-Ah;Sakaguchi, Donald S.
成年大鼠海马祖细胞 (AHPC) 是自我更新的多能神经祖细胞 (NPC)，可以分化为神经元、少突胶质细胞和星形胶质细胞。 AHPC 通过整合素接触周围微环境中的各种分子线索。我们假设整合素受体对于 NPC 很重要。在这项研究中，我们使用免疫细胞化学检查了神经元样、少突胶质细胞样和星形胶质细胞样 AHPC 在支持整合素介导的粘附（层粘连蛋白、纤连蛋白）和不支持整合素介导的粘附（聚-L-鸟氨酸，PLO）的基质上生长时整合素的分布，并表征了铺在层粘连蛋白和纤连蛋白上的 AHPC 的表型分化。粘着斑在镀在纯化基质上的 AHPC 中很突出，但在镀在 PLO 上的 AHPC 中也发现了这种现象。在铺在 PLO 基底上的 AHPC 中观察到的粘着斑可能是通过与内源产生的层粘连蛋白或纤连蛋白的自粘而形成的。我们已经证明，整合素有助于 AHPC 的初始形态分化，因为抑制纤连蛋白与竞争性抑制剂 echistatin 的结合显着减少了处理细胞中存在的突起和微刺的数量，并且还减少了总体细胞面积。最后，我们使用逆转录酶聚合酶链反应表征了 AHPC 中整合素和整合素相关基因子集的遗传特征。这些结果证明了整合素在体外对于 AHPC 的初始形态分化的重要作用。
Adult rat hippocampal progenitor cells (AHPCs) are self-renewing, multipotent neural progenitor cells (NPCs) that can differentiate into neurons, oligodendrocytes, and astrocytes. AHPCs contact a variety of molecular cues within their surrounding microenvironment via integrins. We hypothesize that integrin receptors are important for NPCs. In this study, we have examined the distribution of integrins in neuronal-like, oligodendrocyte-like, and astrocyte-like AHPCs when grown on substrates that support integrin-mediated adhesion (laminin, fibronectin), and those that do not (poly-l-ornithine, PLO) using immunocytochemistry as well as characterized the phenotypic differentiation of AHPCs plated on laminin and fibronectin. Focal adhesions were prominent in AHPCs plated on purified substrates, but were also found in AHPCs plated on PLO. The focal adhesions observed in AHPCs plated on PLO substrates may be formed by self-adhesion to the endogenously produced laminin or fibronectin. We have demonstrated that integrins contribute to the initial morphological differentiation of AHPCs, as inhibition of fibronectin binding with the competitive inhibitor echistatin significantly decreased the number of processes and microspikes present in treated cells, and also decreased overall cell area. Finally, we have characterized the genetic profile of a subset of integrins and integrin-related genes in the AHPCs using reverse transcriptase polymerase chain reaction. These results demonstrate an important role of integrins, in vitro, for the initial morphological differentiation of AHPCs.