Generation of Shox2-Cre allele for tissue specific manipulation of genes in the developing heart, palate, and limb.

Shox2 is expressed in several developing organs in a tissue specific manner in both mice and humans, including the heart, palate, limb, and nervous system. To better understand the spatial and temporal expression patterns of Shox2 and to systematically dissect the genetic cascade regulated by Shox2, we created Shox2-LacZ and Shox2-Cre knock-in mouse lines. We show that the Shox2-LacZ allele expresses beta-galactosidase reporter gene in a fashion that recapitulates the endogenous Shox2 expression pattern in developing organs, including the sinoatrial node (SAN), the anterior portion of the palate, and the proximal region of the limb bud. Conditional deletion of Shox2 in mice carrying the Shox2-Cre allele yielded SAN phenotypes that resemble conventional Shox2 knockout mice. Our results indicate that the Shox2-Cre allele offer a useful tool for tissue specific manipulation of genes in a number of developing organs, particularly in the developing SAN.

Early morphogenesis of the sinuatrial region of the chick heart: a contribution to the understanding of the pathogenesis of direct pulmonary venous connections to the right atrium and atrial septal defects in hearts with right isomerism of the atrial appendages.

The morphogenesis of the sinuatrial region of embryonic hearts is still not well understood. Current matters of dispute are the topogenesis of the future pulmonary vein orifice and the topogenesis of the primary atrial septum. We analyzed the development of the sinuatrial region in chick embryos ranging from Hamburger and Hamilton (HH) stage 14 to 25. Our study disclosed three features of sinuatrial development. First, the primitive atrium of the HH stage 16 chick embryo heart has a separate inflow component. This inflow component takes up the mouth of the confluence of the systemic veins (sinus venosus) as well as the future mouth of the common pulmonary vein (pulmonary pit). The left portion of the atrial inflow component becomes incorporated into the left atrium and its right portion becomes incorporated into the right atrium. Rightward growth of the sinuatrial fold separates the sinus venosus from the left atrium. Second, the pulmonary pit originally forms as a bilaterally paired structure. Its left and right portions are connected to the left and right portions of the atrial inflow component, respectively. Normally, only the left portion of the pulmonary pit deepens to form the common pulmonary vein orifice, whereas the right portion disappears. Third, the primary atrial septum of the chick heart is not formed at the original midline of the embryonic heart, but is formed to the left of the original midline. This finding is in accord with molecular data suggesting that the primary atrial septum derives from the left heart-forming field. Our findings shed new light on the pathogenesis of direct pulmonary venous connections to the right atrium and atrial septal defects in hearts with right isomerism of the atrial appendages.