Stachurska Emilia, Loboda Agnieszka, Niderla-Bielińska Justyna, Szperl Małgorzata, Juszyński Michał, Jozkowicz Alicja, Dulak Jozef, Ratajska Anna
Department of Pathological Anatomy, Medical University of Warsaw, Warszawa, Poland.
Acta Biochim Pol. 2011;58(1):19-29. Epub 2011 Mar 16.
Cellular retinoic acid binding proteins are considered to be involved in retinoic acid (RA) signaling pathways. Our aim was to compare the expression and localization of cellular retinoic acid binding proteins I and II (CRABP I and II) in embryonic mouse hearts during normal development and after a single teratogenic dose of RA. Techniques such as real-time PCR, RT-PCR, Western blots and immunostaining were employed to examine hearts from embryos at 9-17 dpc. RA treatment at 8.5dpc affects production of CRABP I and II in the heart in the 48-h period. Changes in expression of mRNA for retinaldehyde dehydrogenase II (Raldh2), Crabp1 and Crabp2 genes also occur within the same time window (i.e. 10-11dpc) after RA treatment. In the embryonic control heart these proteins are localized in groups of cells within the outflow tract (OT), and the atrioventricular endocardial cushions. A gradient of labeling is observed with CRABP II but not for CRABP I along the myocardium of the looped heart at 11 dpc; this gradient is abolished in hearts treated with RA, whereas an increase of RALDH2 staining has been observed at 10 dpc in RA-treated hearts. Some populations of endocardial endothelial cells were intensively stained with anti-CRABP II whereas CRABP I was negative in these structures. These results suggest that CRABP I and II are independently regulated during heart development, playing different roles in RA signaling, essential for early remodeling of the heart tube and alignment of the great arteries to their respective ventricles.
细胞视黄酸结合蛋白被认为参与视黄酸（RA）信号通路。我们的目的是比较细胞视黄酸结合蛋白I和II（CRABP I和II）在正常发育的胚胎小鼠心脏以及单次致畸剂量的RA处理后的表达和定位。采用实时PCR、RT-PCR、蛋白质免疫印迹和免疫染色等技术检测9-17日龄胚胎的心脏。在8.5日龄时给予RA处理会影响心脏在48小时内CRABP I和II的产生。视黄醛脱氢酶II（Raldh2）、Crabp1和Crabp2基因的mRNA表达变化也发生在RA处理后的同一时间窗口内（即10-11日龄）。在胚胎对照心脏中，这些蛋白定位于流出道（OT）和房室心内膜垫内的细胞群中。在11日龄的环状心脏心肌中观察到CRABP II有标记梯度，而CRABP I没有；在用RA处理的心脏中，这种梯度消失，而在10日龄的RA处理心脏中观察到RALDH2染色增加。一些心内膜内皮细胞群体被抗CRABP II强烈染色，而CRABP I在这些结构中呈阴性。这些结果表明，CRABP I和II在心脏发育过程中受到独立调节，在RA信号传导中发挥不同作用，这对心脏管的早期重塑以及大动脉与各自心室的对齐至关重要。
