Jósvay Katalin, Winter Zoltán, Katona Róbert L, Pecze László, Marton Annamária, Buhala Andrea, Szakonyi Gerda, Oláh Zoltán, Vizler Csaba
1] Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary [2] Institute of Pharmaceutical Analysis, Faculty of Pharmacy, University of Szeged, Szeged, Hungary.
Institute of Pharmaceutical Analysis, Faculty of Pharmacy, University of Szeged, Szeged, Hungary.
Sci Rep. 2014 Oct 27;4:6776. doi: 10.1038/srep06776.
The B6.Cg-Tg(Thy1-YFP)16Jrs/J transgenic mouse strain, widely used to study neuronal development and regeneration, expresses the yellow fluorescent protein (YFP) in the peripheral nerves and the central nervous system under the control of regulatory sequences of the Thy1 gene. The Thy1 (CD90) cell surface glycoprotein is present on many cell types besides neurons, and is known to be involved in cell adhesion, migration and signal transduction. We hypothesized that Thy1-activating conditions could probably activate the truncated Thy1 regulatory sequences used in the Thy1-YFP construct, resulting in YFP transgene expression outside the nervous system. We demonstrated that the stroma of subcutaneous tumours induced by the injection of 4T1 or MC26 carcinoma cells into BALB/c(Thy1-YFP) mice, carrying the same construct, indeed expressed the YFP transgene. In the tumour mass, the yellow-green fluorescent stromal cells were clearly distinguishable from 4T1 carcinoma cells stably transfected with red fluorescent protein. Local inflammation induced by subcutaneous injection of complete Freund's adjuvant, as well as the experimental wound-healing milieu, also triggered YFP fluorescence in both the BALB/c(Thy1-YFP) and B6.Cg-Tg(Thy1-YFP)16Jrs/J mice, pointing to eventual overlapping pathways of wound-healing, inflammation and tumour growth.
B6.Cg-Tg(Thy1-YFP)16Jrs/J转基因小鼠品系广泛用于研究神经元发育和再生,在Thy1基因调控序列的控制下,在周围神经和中枢神经系统中表达黄色荧光蛋白(YFP)。Thy1(CD90)细胞表面糖蛋白除了存在于神经元外,还存在于许多细胞类型中,并且已知其参与细胞粘附、迁移和信号转导。我们推测,激活Thy1的条件可能会激活Thy1-YFP构建体中使用的截短的Thy1调控序列,从而导致YFP转基因在神经系统外表达。我们证明,将4T1或MC26癌细胞注射到携带相同构建体的BALB/c(Thy1-YFP)小鼠中诱导的皮下肿瘤基质确实表达了YFP转基因。在肿瘤块中,黄绿色荧光基质细胞与稳定转染红色荧光蛋白的4T1癌细胞明显不同。皮下注射完全弗氏佐剂诱导的局部炎症以及实验性伤口愈合环境,也在BALB/c(Thy1-YFP)和B6.Cg-Tg(Thy1-YFP)16Jrs/J小鼠中引发了YFP荧光,表明伤口愈合、炎症和肿瘤生长最终存在重叠途径。