Application Zebrafish Neural Tube Malformations Pcp Pathway Gene Function

Neural tube defects(NTDs) are common and severe congenital malformations that result from failure of neural tube closure during early development, and affecting 1-2/1000 established pregnancies worldwide. In China,the incidence rate was highest in shanxi province,the results as high as 9.7/1000 in 1996,there are still 3.5/1000 in 2004.Their etiology is quite complex involving environmental and genetic factors, their underlying molecular and cellular pathogenic mechanisms remain poorly understood. Previous genetic studies in NTDs have focused mainly on folate metabolism-related genes, recent studies in NTDs mouse models have demonstrated an essential role of the planar cell polarity pathway (PCP) in mediating a morphogenetic process called convergent extension(CE) during neural tube closure.Alterations in members of planar cell polarity pathway lead to neural tube defect in vertebrate models, representing novel candidate gene for human neural tube defect.The research focus on the planar cell polarity pathway genes Vang12 and Dishevelled (Dsh/Dvl),which have a crucial role in neural tube close as the planar cell polarity pathway genes. Vangl was first recognised in Drosophila and named Van Gogh(Vang).Vang12 (Van Gogh-like 2) is a mammalian homolog of the Drosophila gene Van Gogh(Vang), also known as Strabismus (Stbm),which has been shown highly conserved in evolution.In this study,we use the two mutations of Vang12 from the human neural tube defects family,which have been found it will change the amino acid sequence and influence the interation with Dishevelled. This work constructed a series of transgenic vectors,such as Vang12 marked GFP or Mcherry constructed to pEGFP-C1 and PME-MCS,for investgating the role of Vang12 in neural tube defects.The Dishevelled expression pattern of planar cell polarity pathway gene was obtained by RT-PCR and whole mount in situ hybridization (WISH) and PCR sequencing and nucleotide and protein sequence was analyzed by bioinformaties.The results of investigations are offered as follows:1. The work constructed transgenic vectors of Vang12 marked red or green fluorescence from planar cell polarity pathway genes which for studying the mutation of Vang12 in the function of neural tube close.2. This findings are the first to provie the expression pattern of Dishevelled (Dvl1/Dvl2/ Dvl3) from planar cell polarity pathway genes in zebrafish embryo,these study provide the foundation for the future research in the planar cell polarity pathway in zebrafish.3. The work did the further study for the interation between planar cell polarity pathway and neural tube defects,which gradually established a perfect platform for the study of neural tube defects and provide the new theory and detection technology in zebrafish model.This work is the firs report about Vangl2 and Dishevelled of zebrafish,which enriched the zebrafish planar cell polarity pathway genes,Combined with Previous studies on the results of planar cell polarity pathway, it is believed that it provide the foundation for further studying of planar cell polarity pathway gene in the zebrafish neural tube formation.