Analysis Of Regulation Of Protein Phosphatase-2Acβ In Fish Embryos Development

Reversible phosphorylation of proteins are important biological events, which are mainly involved in the regulation of DNA damage, cell proliferation, metabolism and even cancer in eukaryotes. Reversible phosphorylation of proteins are catalyticed by protein kinase and protein phosphatase. Protein Phosphatase-2A (Protein Phosphatase-2A.PP-2A) is one of the most abundant protein phosphatase in animal body.The holoenzyme for PP-2A consists of a catalytic subunit C, a regulatory subunit B and a structural subunit A. The catalytic subunit is the basic structure of the enzyme, however, PP-2Ac are carrying two isomers: PP-2Aca and PP-2Acβ.This article is mainly focused on PP-2Acβ, one isoform of PP-2Ac. we isolated the total RNAs from developing embryos at stages of2-cell, multiple-cell, blastula, gastrula, neurula, optical vesicle, brain differentiation, muscle differentiation, heart beat, eye pigmentation, body pigmentation and hatching larvae and nine tissues (liver, spermary, ovary, brain, kidney, heart, muscle, gill and fin) of goldfish. Conducted the differential expression pattern of the catalytic subunits β of Protein Phosphatase-2A (PP-2A) in twelve development stages embryos of goldfish and nine kinds of tissues of goldfish using RT-PCR. We revealed:1) The mRNA level for PP-2Acβ were expressed at the highest level in the period of optical vesicle. The higher levels in the period of muscle differentiation, multiple cell, blastula, gastrula, eye pigment, hatching larvae, the low levels in the period of two cell, neurula, body pigment, the lowest levels in the period of brain differentiation and heart beat.2)the expression levels of PP-2Acβ were different in nine kinds of tissues of the goldfish. The mRNA level for PP-2Acβ were expressed at the highest level in ovary. The higher levels in heart, muscle, brain, fin, the low levels in kidney, liver, gill, and ovary. It can be speculated that the gene of PP-2Aβ may play an important role in different stages of embryonic development of goldfish and nine kinds of tissues of goldfish.PP2Acβ-MO were injected into each fertilized egg by Microinjection to inhibit the activity of PP-2Acβ, so we can analysis the functional mechanism of PP-2AcP in twelve development stages embryos of goldfish. We observed that the successful injection of goldfish embryos have serious abnormalities, for example, differentiation pattern change due to abnormal differentiation of embryonic eye and unequal cell division due to embryonic spinal bends, which further highlighted the PP-2Acβ did play an important role in the normal development of the embryo. We further observed organizational change of the successful injection of embryos by paraffin sectioning technique, it turned out that the development of retina and lens were abnormal, with malformation of the brain.Embryos generated a variety of obvious deformity after suppressed the expression of PP-2Acβ by microinjection MO. It was indicated that PP-2Ac was one of the important Protein Phosphatase in goldfish and played an important role in regulating embryonic development of goldfish.