Expression And Function Analysis Of Rho-associated Protein Kinase2in Dujesia Japonica

Rho kinase2(Rho-associated protein kinase2, Rock2) is a downstream target effector molecule Rho Rock of a subtypes; Rock2extensively involved in regulating cell contraction, movement, proliferation, differentiation and apoptosis processes, inhibition Rock2can significantly relieve cancer and cardiovascular disease. Rock2mainly expressed in the brain and nerve tissue, nerve damage can be adjusted after the synaptic regeneration.N425clone was obtained from the cDNA library of Dugesia japonica. The sequence analysis showed that the largest ORF consisting734bp which encoded a deduced protein of142amino acid. Sequence alignment by NCBI BLASTP program, and37%similarity paramecium, We named DjRock2gene in this study.Using Adult individuals in situ hybridization and RT-PCR detection of the expression of genes in planarians DjRock2regeneration process.When reproducing three days and five days, without regeneration of the head and tail, the positive signal is weak. Only a positive signal position in the nervous system, the signal also in regeneration wound.10days later, DjRock2genes specifically expressed in the nervous still. The result of Immunohistochemistry showed that DjRock2protein and DjRock2gene at the same location. DjRock2genes specifically expressed in the nervous system showed that, DjRock2genes involved in the maintenance of the nervous system. Expression was significantly in the head, showed that DjRock2genes play a key role in the optic nerve of the formation and maintenance. Expression description in the wound showed that, DjRock2genes involved in cell proliferation and differentiationFor the first time, DjRock2gene was inserted into L4440vector. Through DjRock2interference test showed that, knock down the DjRock2gene be determined, knock down DjRock2lead to lost regenerative capacity, and affect the life-sustaining of normal cells. This is consistent with the function of the Rock2protein. Through observation, regeneration is inhibited, and a large number of cell death on the wound. Speculate DjRock2gene and the differentiation of adult stem cells showed a positive correlation. Through the RT-PCR technique to detect the relationship between the phenotypic changes and DjRock2interference, to be sure DjRock2involved in planarian regeneration process. Inference, this may be by regulating the Rock classic signaling pathway, and regulate MLC. Control versatile adult stem cell proliferation and differentiation, to achieve the purpose of regulating regeneration. To establish the foundation for the research of Rock2gene.