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丝氨酸/苏氨酸激酶33(Stk33)与波形蛋白在下丘脑中的共定位。

Co-localization of serine/threonine kinase 33 (Stk33) and vimentin in the hypothalamus.

作者信息

Brauksiepe Bastienne, Baumgarten Lisa, Reuss Stefan, Schmidt Erwin R

机构信息

Institute of Molecular Genetics, Johannes Gutenberg-University Mainz, Johann-Joachim Becherweg 32, 55128, Mainz, Germany.

出版信息

Cell Tissue Res. 2014 Jan;355(1):189-99. doi: 10.1007/s00441-013-1721-8. Epub 2013 Sep 22.

Abstract

We investigate the immunoreactivity of serine/threonine kinase 33 (Stk33) and of vimentin in the brain of mouse, rat and hamster. Using a Stk33-specific polyclonal antibody, we show by immunofluorescence staining that Stk33 is present in a variety of brain regions. We found a strong staining in the ependymal lining of all cerebral ventricles and the central canal of the spinal cord as well as in hypothalamic tanycytes. Stk33 immunoreactivity was also found in circumventricular organs such as the area postrema, subfornical organ and pituitary and pineal glands. Double-immunostaining experiments with antibodies against Stk33 and vimentin showed a striking colocalization of Stk33 and vimentin. As shown previously, Stk33 phosphorylates recombinant vimentin in vitro. Co-immunoprecipitation experiments and co-sedimentation assays indicate that Stk33 and vimentin are associated in vivo and that this association does not depend on further interacting partners (Brauksiepe et al. in BMC Biochem 9:25, 2008). This indicates that Stk33 is involved in the dynamics of vimentin polymerization/depolymerization. Since in tanycytes the vimentin expression is regulated by the photoperiod (Kameda et al. in Cell Tissue Res 314:251-262, 2003), we determine whether this also holds true for Stk33. We study hypothalamic sections from adult Djungarian hamsters (Phodopus sungorus) held under either long photoperiods (L:D 16:8 h) or short photoperiods (L:D 8:16 h) for 2 months. In addition, we examine whether age-dependent changes in Stk33 protein content exist. Our results show that Stk33 in tanycytes is regulated by the photoperiod as is the case for vimentin. Stk33 may participate in photoperiodic regulation of the endocrine system.

摘要

我们研究了丝氨酸/苏氨酸激酶33(Stk33)和波形蛋白在小鼠、大鼠和仓鼠大脑中的免疫反应性。使用Stk33特异性多克隆抗体,我们通过免疫荧光染色显示Stk33存在于多种脑区。我们在所有脑室的室管膜内衬、脊髓中央管以及下丘脑的伸长细胞中发现了强烈的染色。在诸如最后区、穹窿下器官、垂体和松果体等室周器官中也发现了Stk33免疫反应性。用抗Stk33和波形蛋白的抗体进行的双重免疫染色实验显示,Stk33和波形蛋白有显著的共定位。如先前所示,Stk33在体外可使重组波形蛋白磷酸化。免疫共沉淀实验和共沉降分析表明,Stk33和波形蛋白在体内相互关联,且这种关联不依赖于其他相互作用的伙伴(Brauksiepe等人,《BMC生物化学》9:25,2008年)。这表明Stk33参与了波形蛋白聚合/解聚的动态过程。由于在伸长细胞中波形蛋白的表达受光周期调节(Kameda等人,《细胞与组织研究》314:251 - 262,2003年),我们确定这对Stk33是否也成立。我们研究了在长光周期(L:D 16:8小时)或短光周期(L:D 8:16小时)下饲养2个月的成年黑线毛足鼠(Phodopus sungorus)的下丘脑切片。此外,我们检查了Stk33蛋白含量是否存在年龄依赖性变化。我们的结果表明,伸长细胞中的Stk33与波形蛋白一样受光周期调节。Stk33可能参与内分泌系统的光周期调节。

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