Nervous System Related Gene Expression And Function Analysis

On the basis of previous experimental work, our group found that protein4.1family members can interact with the members of the Nectin-like family and directly impact on their biological function. Protein4.1family consist of five members:4.IN,4.1B,4.1R4.1G and4.1DAL (a shorter transcript of4.1B). To be a cytoskeletal protein family, protein4.1family members can connect membrane proteins (including adhesion molecules, receptors and ion channels) with intracellular proteins (including cytoskeletal proteins, signaling molecules or transcription factors), and form a functional protein complex to regulate the molecular events of downstream. From the structural perspective, the membrane binding domain (MBD) whose molecular weight is30kDa is essential for the4.1family members to interact with the intracellular protein. There are three domains which are shared by all the family members:the N-terminal membrane-binding domain (MBD), the ghost protein and actin-binding domain (SABD) and the conservative carboxyl-terminal domain (CTD).We focus on4.IN,4.1R, and4.1B because they are found to express in the nervous system. Among them,4.1N and4.1B are in the brain-specific expression, while the4.1R expresses widely, that also express in some specific groups of neurons. Constructing three gene-specific probe, we observe the expression pattern of the three members in the brain at the different embryonic and postnatal stage by in situ hybridization. Combined with the expression of Nectin-like family members in the same space-time point, it can provide us the direct clues and evidence for studying the relationship of their biological functionAll-trans retinoic acid is an anti-tumor drug and has been widely used in clinical medicine. It can induce tumor cell apoptosis and differentiation effectivly, and also be found of having a therapeutic effect on skin diseases in clinical. However, it will increase the risk of neural tube defects if used during pregnancy. At present, researchers found that we can get the disease animal model which is similar to the human phenotype of neural tube defect by all trans retinoic acid induction, that treat the mouse embryos at special time point. It provides us a powerful means to study the hazard of NTD.In our study, we used the all-trans retinoic acid as a inducer to treat the pregnant ICR strain mice at special time point so as to build the disease animal model for studying the molecular mechanism of NTD. Combined with microarray analysis data, we focus on four genes:ASH2L, HDAC4, NSPC1and DOK5, whose changes of expression are obvious compared with the control group. So their changes in the level of transcription and translation were studied.By the technology of Real-time PCR and Western Blotting, we measured these changes in mRNA and protein levels, and after a corresponding statistical analysis, found that the four genes showed obviously down-regulation in the nervous system of treatment group compared with the control.The results suggest that ASH2L, HDAC4, NSPC1and DOK5may be the underlying genes of inhibiting all trans RA-induced neural tube defects. The study has laid the foundation to reveal theirs biological function in the pathogenic process of neural tube defects.