Herrfurth Leonard, Theis Verena, Matschke Veronika, May Caroline, Marcus Katrin, Theiss Carsten
Medizinische Fakultät, Institut für Anatomie, Abteilung für Cytologie, Ruhr-Universität Bochum Bochum, Germany.
Abteilung für Medizinische Proteomik/Bioanalytik, Medizinisches Proteom-Center, Ruhr-University Bochum Bochum, Germany.
Front Mol Neurosci. 2017 Jan 30;10:2. doi: 10.3389/fnmol.2017.00002. eCollection 2017.
Vascular endothelial growth factor (VEGF) is well known as the growth factor with wide-ranging functions even in the central nervous system (CNS). Presently, most attention is given to the investigation of its role in neuronal protection, growth and maturation processes, whereby most effects are mediated through VEGF receptor 2 (VEGFR-2). The purpose of our current study is to provide new insights into the impact of VEGF on immature and mature Purkinje cells (PCs) in accordance with maturity and related receptor expression. Therefore, to expand our knowledge of VEGF effects in PCs development and associated VEGFR-2 expression, we used cultivated organotypic cerebellar slice cultures in immunohistochemical or microinjection studies, followed by confocal laser scanning microscopy (CLSM) and morphometric analysis. Additionally, we incorporated in our study the method of laser microdissection, followed by quantitative polymerase chain reaction (qPCR). For the first time we could show the age-dependent VEGF sensitivity of PCs with the largest promoting effects being on dendritic length and cell soma size in neonatal and juvenile stages. Once mature, PCs were no longer susceptible to VEGF stimulation. Analysis of VEGFR-2 expression revealed its presence in PCs throughout development, which underlined its mediating functions in neuronal cells.
血管内皮生长因子(VEGF)是一种广为人知的生长因子,即使在中枢神经系统(CNS)中也具有广泛的功能。目前,人们主要关注其在神经元保护、生长和成熟过程中的作用,其中大多数效应是通过VEGF受体2(VEGFR-2)介导的。我们当前研究的目的是根据成熟度和相关受体表达,深入了解VEGF对未成熟和成熟浦肯野细胞(PCs)的影响。因此,为了拓展我们对VEGF在PCs发育及相关VEGFR-2表达方面作用的认识,我们在免疫组织化学或显微注射研究中使用了培养的小脑器官型切片培养物,随后进行共聚焦激光扫描显微镜(CLSM)和形态计量分析。此外,我们在研究中纳入了激光显微切割方法,随后进行定量聚合酶链反应(qPCR)。我们首次能够证明PCs对VEGF的敏感性存在年龄依赖性,在新生期和幼年期对树突长度和细胞体大小的促进作用最大。一旦成熟,PCs就不再对VEGF刺激敏感。对VEGFR-2表达的分析显示其在PCs整个发育过程中均有存在,这突出了其在神经元细胞中的介导功能。