Sel Saadettin, Münzenberg Christoph, Nass Norbert, Kalinski Thomas, Datan Maja, Auffarth Gerd U, Töteberg-Harms Marc, Zenkel Matthias, Kruse Friedrich E, Paulsen Friedrich, Schicht Martin
Department of Ophthalmology, University Erlangen-Nuremberg, Erlangen, Germany.
Gene Expr Patterns. 2013 Oct;13(7):280-6. doi: 10.1016/j.gep.2013.05.003. Epub 2013 May 25.
The forkhead transcription factor Foxk1 is an important regulator of myogenic progenitor cells. Since our previous data from mouse retina revealed that Foxk1 is upregulated in Ptf1a-deficient mice we investigated the spatial and temporal expression of Foxk1 during development of mouse retina. Expression of Foxk1 was analyzed on both mRNA and protein level. To identify Foxk1 transcripts, retina and cerebrum (positive control) of adult animals (postnatal day 90 (P90)) was subjected to reverse transcription polymerase chain reaction (RT-PCR) and sequencing of the amplified cDNA. The Foxk1 protein was analyzed in adult retina by Western blotting and in developing eyes at embryonic day (E) 13, 15, E17, P0, P4, P7, P10 and P90 by immunohistochemistry. Localization of Foxk1 expression was determined using cell-specific markers by double labelling. Foxk1 transcripts were detected in adult retina by RT-PCR and confirmed by sequencing. Western blot analysis confirmed the expression of Foxk1 protein in the adult retina. Immunohistochemical examination of developing eyes localized the protein to bipolar, amacrine and ganglion cells with an onset of Foxk1 expression from E15 onwards. The expression pattern during development suggests that Foxk1 may have an important role in retinal cells.
叉头转录因子Foxk1是肌源性祖细胞的重要调节因子。鉴于我们之前从小鼠视网膜获得的数据显示Foxk1在Ptf1a基因缺陷小鼠中上调，我们研究了Foxk1在小鼠视网膜发育过程中的时空表达。从mRNA和蛋白质水平对Foxk1的表达进行了分析。为了鉴定Foxk1转录本，对成年动物（出生后第90天（P90））的视网膜和大脑（阳性对照）进行逆转录聚合酶链反应（RT-PCR）以及对扩增的cDNA进行测序。通过蛋白质印迹法分析成年视网膜中的Foxk1蛋白，并通过免疫组织化学分析胚胎第13天、15天、17天、出生后第0天、第4天、第7天、第10天和第90天发育中的眼睛中的Foxk1蛋白。通过使用细胞特异性标志物进行双重标记来确定Foxk1表达的定位。通过RT-PCR在成年视网膜中检测到Foxk1转录本，并通过测序得以证实。蛋白质印迹分析证实了Foxk1蛋白在成年视网膜中的表达。对发育中的眼睛进行免疫组织化学检查发现，该蛋白定位于双极细胞、无长突细胞和神经节细胞，Foxk1的表达从胚胎第15天开始。发育过程中的表达模式表明Foxk1可能在视网膜细胞中发挥重要作用。
