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“Loop-tail (Lp) mice show a very sever e neural tube defect (craniorachischists) caused by mutations in the Vangl2 tame (D255E, S464N) Mammalian Vangl1 and Vangl2 are membrane proteins that play critical roles in development such as establishment of planar cell polarity (PCP) in epithelial layers and convergent extension movements during neurogenesis and cardiogenesis. Vangl proteins are thought to assemble with other PCP proteins (Dvl, Pk) to form membrane-bound PCP

signaling complexes that provide polarity information to the cell. In the present study, we show that Vangl I is expressed exclusively at the plasma membrane of transfected MDCK cells, where it is targeted to the basolateral membrane Experiments with an inserted exofacial HA epitope indicate that the segment delimited by the predicted transmembrane domains 1 and 2 is exposed to the extracellular milieu Comparative MRT67307 inhibitor studies of the Lp-associated

pathogenic mutation D255E indicate that the targeting selleck kinase inhibitor of the mutant variant at the plasma membrane is greatly reduced; the mutant variant is predominantly retained intracellularly in endoplasmic reticulum (ER) vesicles colocalizing with the ER marker calreticulin. In addition, the D255E variant shows drastically reduced stability with a half-life of similar to 2 h, compared to > 9 h for its wild type counterpart and is rapidly degraded in a proteasome-dependent and MG132 sensitive pathway. These (inclines highlight a critical role for D255 for normal folding and processing of Vangl proteins, with highly conservative substitutions not tolerated at that site. Our study provide an experimental framework for the analysis of human VANGL mutations recently identified in familial and sporadic cases of spina bifida.”
“Heart disease is the leading cause of death in the United States. Recent studies demonstrate that fetal programming of PKC epsilon gene repression results in ischemia-sensitive phenotype in the heart. The present study tests the MK-2206 concentration hypothesis that increased norepinephrine causes epigenetic repression of PKC epsilon gene in the heart

via Nox1-dependent reactive oxygen species (ROS) production. Prolonged norepinephrine treatment increased ROS production in fetal rat hearts and embryonic ventricular myocyte H9c2 cells via a selective increase in Nox1 expression. Norepinephrine-induced ROS resulted in an increase in PKC epsilon promoter methylation at Egr-1 and Sp-1 binding sites, leading to PKC epsilon gene repression. N-acetylcysteine, diphenyleneiodonium, and apocynin blocked norepinephrine-induced ROS production and the promoter methylation, and also restored PKC epsilon mRNA and protein to control levels in vivo in fetal hearts and in vitro in embryonic myocyte cells. Accordingly, norepinephrine-induced ROS production, promoter methylation, and PKC epsilon gene repression were completely abrogated by knockdown of Nox1 in cardiomyocytes.