Characterization of Diaphanous-related formin FMNL2 in human tissues.
Characterization of Diaphanous-related formin FMNL2 in human tissues.
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DOI：
10.1186/1471-2121-11-55
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发表时间：
2010-07-15
期刊：
影响因子：
--
通讯作者：
Carpén O
中科院分区：
文献类型：
--
作者：
Gardberg M;Talvinen K;Kaipio K;Iljin K;Kampf C;Uhlen M;Carpén O
透明相关的formins管理肌动蛋白为基础的过程中涉及许多细胞功能，如细胞运动和入侵。与恶性表型相关的发育过程和细胞变化的可能联系使其成为有趣的研究目标。尽管如此，我们对哺乳动物的鳃的组织分布和细胞位置知之甚少。在这里，我们已经进行了一个全面的分析，在人体组织中的femtocell家族成员femtocell样2（FMNL 2）。提出并表征FMNL 2抗体。亲和纯化的FMNL 2抗体通过Western印迹、北方印迹、肽竞争测定和siRNA实验进行验证。基于生物信息学的mRNA分析表明FMNL 2在人体组织中广泛表达。在中枢和外周神经系统中观察到最高的mRNA水平。对26种不同人体组织的免疫组织化学分析表明，FMNL 2广泛表达，与mRNA谱一致。在中枢神经系统中检测到最广泛的表达，因为神经元和神经胶质细胞都表达FMNL 2。在许多上皮细胞中也观察到强表达。然而，在不同细胞类型中的表达不是普遍存在的。许多间充质细胞类型表现出弱免疫反应性，在许多组织中可见缺乏表达的细胞。FMNL 2的亚细胞定位在细胞质中，在某些组织中检测到强烈的核周点。在培养的细胞中，FMNL 2主要表现为细胞质定位与高尔基体一致的核周积累。此外，FMNL 2与F-肌动蛋白共定位于WM 164人黑素瘤细胞中细胞突起的尖端。这一发现与FMNL 2提出的在变形细胞迁移期间在细胞突起中形成肌动蛋白丝的功能一致。FMNL 2不仅在中枢神经系统中表达，而且在多种人体组织中表达。在胃肠道和乳腺上皮、淋巴组织、胎盘和生殖道中表达尤其强烈。在培养的黑色素瘤细胞中，FMNL 2与细胞突起尖端的F-肌动蛋白点共定位。
Diaphanous-related formins govern actin-based processes involved in many cellular functions, such as cell movement and invasion. Possible connections to developmental processes and cellular changes associated with malignant phenotype make them interesting study targets. In spite of this, very little is known of the tissue distribution and cellular location of any mammalian formin. Here we have carried out a comprehensive analysis of the formin family member formin -like 2 (FMNL2) in human tissues. An FMNL2 antibody was raised and characterized. The affinity-purified FMNL2 antibody was validated by Western blotting, Northern blotting, a peptide competition assay and siRNA experiments. Bioinformatics-based mRNA profiling indicated that FMNL2 is widely expressed in human tissues. The highest mRNA levels were seen in central and peripheral nervous systems. Immunohistochemical analysis of 26 different human tissues showed that FMNL2 is widely expressed, in agreement with the mRNA profile. The widest expression was detected in the central nervous system, since both neurons and glial cells expressed FMNL2. Strong expression was also seen in many epithelia. However, the expression in different cell types was not ubiquitous. Many mesenchymal cell types showed weak immunoreactivity and cells lacking expression were seen in many tissues. The subcellular location of FMNL2 was cytoplasmic, and in some tissues a strong perinuclear dot was detected. In cultured cells FMNL2 showed mostly a cytoplasmic localization with perinuclear accumulation consistent with the Golgi apparatus. Furthermore, FMNL2 co-localized with F-actin to the tips of cellular protrusions in WM164 human melanoma cells. This finding is in line with FMNL2's proposed function in the formation of actin filaments in cellular protrusions, during amoeboid cellular migration. FMNL2 is expressed in multiple human tissues, not only in the central nervous system. The expression is especially strong in gastrointestinal and mammary epithelia, lymphatic tissues, placenta, and in the reproductive tract. In cultured melanoma cells, FMNL2 co-localizes with F-actin dots at the tips of cellular protrusions.
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影响因子：
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