SUMMARYEndothelial tubulogenesis is a crucial step in the formation of functional blood vessels during angiogenesis and vasculogenesis. Here, we use in vivo imaging of living zebrafish embryos expressing fluorescent fusion proteins of b-Actin, a-Catenin, and the ERM family member Moesin1 (Moesin a), to define a novel cord hollowing process that occurs during the initial stages of tubulogenesis in intersegmental vessels (ISVs) in the embryo. We show that the primary lumen elongates along cell junctions between at least two endothelial cells during embryonic angiogenesis. Moesin1-EGFP is enriched around structures that resemble intracellular vacuoles, which fuse with the luminal membrane during expansion of the primary lumen. Analysis of silent heart mutant embryos shows that initial lumen formation in the ISVs is not dependent on blood flow; however, stabilization of a newly formed lumen is dependent upon blood flow. Zebrafish moesin1 knockdown and cell transplantation experiments demonstrate that Moesin1 is required in the endothelial cells of the ISVs for in vivo lumen formation. Our analyses suggest that Moesin1 contributes to the maintenance of apical/basal cell polarity of the ISVs as defined by adherens junctions. Knockdown of the adherens junction protein Ve-cadherin disrupts formation of the apical membrane and lumen in a cell-autonomous manner. We suggest that Ve-cadherin and Moesin1 function to establish and maintain apical/basal polarity during multicellular lumen formation in the ISVs.

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We address the problem of constructing and identifying a valid joint probability density function from a set of specified conditional densities. The approach taken is based on the development of relations between the joint and the conditional densities using Markov random fields (MRFs). We give a necessary and sufficient condition on the support sets of the random variables to allow these relations to be developed. This condition, which we call the Markov random field support condition, supercedes a common assumption known generally as the positivity condition. We show how these relations may be used in reverse order to construct a valid model from specification of conditional densities alone. The constructive process and the role of conditions needed for its application are illustrated with several examples, including MRFs with multiway dependence and a spatial beta process. Academic Press

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