Studies On The Function Of Human Disease-associated Genes Palladin And Gpr110 And The Underlying Mechanism

Part ? The mechanism study of Palladin on the process of neural tube closureActin cytoskeleton-associated protein palladin plays an important role in cell motility,morphogenesis and adhesion.Palladin deficient mouse embryos are lethal before embryonic day(E)15.5,and exhibit severe cranial neural tube and body wall closure defects.To further understand how palladin regulates the process of cranial neural tube closure(NTC),we first focused on the expression pattern of palladin and found that palladin is predominantly expressed in the neural folds at E9.5.We further checked the major cellular events of neural epithelium in the neural folds that contribute to NTC during the early embryogenesis.Palladin deficiency leads to a disturbance of cytoskeleton in the neural tube and the cultured neural progenitors.Conformably,cytoskeleton related proteins p-cofilin,?-actinin and argbp2 are down-regulated in Palladin-/-embryonic brains compared with wild type(wt)counterparts.Furthermore,increased cell proliferation,decreased cell differentiation and diminished apical cell apoptosis of neural epithelium are found in palladin deficient embryos.Cell cycle of neural progenitors in Palladin-/-embryos is much shorter than that in wt ones.In consistent with these findings,the expression levels of Cyclin E and Bcl-2 are increased in Palladin-/-brain tissues.Unexpectedly,cell adhesion shows no significant changes in Palladin-/-neural folds.Taken together,our data suggest that palladin is expressed in the proper spatio-temporal pattern in the neural folds.It plays a crucial role in regulating mouse cranial NTC by modulating cytoskeleton,proliferation,differentiation,and apoptosis of neural epithelium.Our findings facilitate further study of the function of palladin and the underlying molecular mechanism involved in NTC.Part ? Generation of Gpr110 knockout mice and phonotype ana lysis of Gpr110-/-miceGpr110 is an adhesion GPCR,which is highly expressed in human prostate cancer and lung cancer,so it is defined as an oncogene.In order to study the function of Gpr110 in vivo,we constructed Gpr110 gene knockout mice.Both heterozygous and homozygous mice were normal in body weight.In offspring from heterozygote cross with heterozygote,the sex and genotype distribution are both normal.Gpr110 gene was mainly expressed in prostate and kidney in mice.HE staining showed that the epithelial cells of 18-month old Gpr110-/-prostate were degenerated and shed.Decreased adhesion was observes in Gpr110-/-prostate epithelial cells.In consistent with these findings,the expression of A4Annexin and Osbpl3was significantly decreased in Gpr110-/-prostate.Urea nitrogen and creatinine were greatly increased in Gpr110-/-serum.mRNA array showed the expression level of an IgA nephropathy susceptible gene St6galnac2 was 120 times higher expressed in Gpr110-/-prostate than that in wt.HE staining showed an increased thickness of mesangial in Gpr110-/-glomerular.Transmission electron microscopy of kidney showed an accumulation of electron dense substance in Gpr110-/-glomerular.Taken together,we constructed Gpr110 knockout mice.The phenotype of homozygous mice was analyzed on transcriptomics level.Our results showed that Gpr110 plays a role in maintaining the structure and function of mouse prostate and kidney.Our findings facilitate further study of Gpr110 in the future,and may even provide a new solution for the treatment of IgA nephropathy.