The Expression Of Phenylalanine Hydroxylase In The Central Nervous System Of Ragworm Neanthes Japonica (Polychaeta, Annelida)

ObjectiveUsing three experimental methods,to provide the evidence that phenylalanine hydroxylase in the central nervous system of ragworm Neanthes japonica in protein and nucleic acid levels,which provide the foundation for further study of aromatic amino acid hydroxylase gene evolution in invertebrate.Methods1.Using Western blot method,detecting the phenylalanine hydroxylase protein whether expressed in the central nervous system of ragworm Neanthes japonica.2.Making paraffin sections and using immunohistochemical technique,to identify the presence and distribution of the phenylalanine hydroxylase gene in the central nervous system of Neanthes japonica.3.Clone PAH partial gene fragments from the central nervous system of Neanthes japonica by RT-PCR,constructing recombinant plasmid and picking a single positive clones,sequencing and comparing homology with other species PAH gene.Results1.Western blot marked a weight of 52 k Da molecular,indicating that the PAH is present in Neanthes japonica central nervous system.2.The immunohistochemical results showed that the positive phenylalanine hydroxylase protein expressed mainly in abdominal,dorsal and sides of forebrain and midbrain and ventral nerve cord.3.The PAH gene cloning by RT-PCR results showed that the phenylalanine hydroxylase exist in the central nervous system of Neanthes japonica.4.The sequence of cloned PAH segment has homology with others species by BLAST.ConclusionsThree kinds of experimental methods proved that PAH is present in the lower invertebrate organisms Neanthes japonica,and have a high homology with a variety of animals.