Colas J F, Lawson A, Schoenwolf G C
Department of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City 84132, USA.
Dev Dyn. 2000 Jun;218(2):316-30. doi: 10.1002/(SICI)1097-0177(200006)218:2<316::AID-DVDY6>3.0.CO;2-8.
Heart development in the chick embryo proceeds from bilateral mesodermal primordia established during gastrulation. These primordia migrate to the midline and fuse into a single heart trough. During their migration as a cohesive sheet, the cells of the paired heart fields become epithelial and undergo cardiac differentiation, exhibiting organized myofibrils and rhythmic contractions near the time of their fusion. Between the stages of cardiomyoblast commitment and overt differentiation of cardiomyocytes, a significant time interval exists. Using a new riboprobe (usmaar) for whole-mount in situ hybridization in chick embryos, we report the earliest phases of smooth muscle alpha-actin (smaa) mRNA distribution during the precontractile developmental window. We show that ingressed heart-forming regions express smaa by the head-process stage (Hamburger and Hamilton stage 5). In addition, we used usmaar to study the formation and early morphogenesis of the heart. Consistent with fate mapping studies (Garcia-Martinez and Schoenwolf [1993] Dev. Biol. 159:706-719; Schoenwolf and Garcia-Martinez [1995] Cell Mol. Biol. Res. 41:233-240; Garcia-Martinez et al., in preparation), our results with this probe, combined with detailed histological and SEM analyses of the so-called cardiac crescent, demonstrate unequivocally that the heart arises from separated and paired heart rudiments, rather than from a single crescent-shaped rudiment (that is, prior to fusion of the paired heart rudiments to establish the straight-heart tube, the rostral midline of the cardiac crescent lacks mesodermal cells and consequently fails to label with usmaar). Smaa is also expressed in the splanchnic and somatic mesoderm, marking the earliest step in coelom formation. Consequently, we also used usmaar to describe formation of the pericardium. Finally, we provide evidence of a post-transcriptional level of control of smaa gene expression in the heart fields. Our results suggest that the expression of smaa may mark a primitive mesodermal state from which definitive cell types can be derived through inductive events.
鸡胚心脏发育始于原肠胚形成期建立的双侧中胚层原基。这些原基迁移至中线并融合形成单一的心管。在作为一个紧密相连的薄片迁移过程中,成对心脏区域的细胞转变为上皮细胞并经历心脏分化,在融合时显示出有组织的肌原纤维和节律性收缩。在心肌母细胞定向分化为明显的心肌细胞阶段之间,存在显著的时间间隔。利用一种新的核糖核酸探针(usmaar)对鸡胚进行全胚胎原位杂交,我们报告了收缩前发育窗口期间平滑肌α-肌动蛋白(smaa)mRNA分布的最早阶段。我们发现,在头突期(汉伯格和汉密尔顿第5期),迁入的心脏形成区域表达smaa。此外,我们利用usmaar研究心脏的形成和早期形态发生。与命运图谱研究结果一致(加西亚-马丁内斯和舍恩沃尔夫[1993]《发育生物学》159:706 - 719;舍恩沃尔夫和加西亚-马丁内斯[1995]《细胞与分子生物学研究》41:233 - 240;加西亚-马丁内斯等人,正在准备中),我们使用该探针的结果,结合对所谓心脏新月区的详细组织学和扫描电镜分析,明确证明心脏起源于分离且成对的心脏原基,而非单个新月形原基(即,在成对心脏原基融合形成直心管之前,心脏新月区的头侧中线缺乏中胚层细胞,因此无法用usmaar标记)。Smaa也在内脏和体壁中胚层表达,标志着体腔形成的最早步骤。因此,我们还利用usmaar描述心包的形成。最后,我们提供了心脏区域中smaa基因表达在转录后水平受到调控的证据。我们的结果表明,smaa的表达可能标志着一种原始中胚层状态,通过诱导事件可从中衍生出确定的细胞类型。