Scr Effects Of UVB Irradiation On Protein Expression Of Early Embryos Of Bufo Bufo Gargarizans

Ultraviolet(UV) is a non-ionizing radiation photophysical factor,and ultraviolet radiation B(UVB) has a very important role on organisms.In order to study the effects of UVB on early development embryos of amphibians,we use early embryos of Bufo bufo gargarizans as experimental material.Embryos at different developmental stages were irradiated by 302 nm UVB radiation.The changes of protein expression of embryos were studied on development patterns,protein expression and organization levels before and after UVB irradiation.1) Different stages of Bufo bufo gargarizans's embryos were irradiated by 304μw/cm~2 UVB.Then we observed the development of the embryos and calculated the livability and abnormal rate.The results indicated that,the effects of UVB on hatching rate was not remarkable;the livability of the embryos changed anomalous after treated by UVB; the abnormal rate changed anomalous during the earlier stages,but rised evidently at the later stages.2) Total protein of blastulas and neural embryos before and after UVB irradiation were separated by immobilized pH gradient based two dimensional gel electrophoresis(2-DE).After colloidal Coomassie brilliant staining,expression proteins were analyzed by ImageMaster 2D Platinum 6.0 image analysis software. The results showed that,UVB irradiation affected significantly on the amount and distribution of embryonic proteins:in blastula,the expression of 27 protein spots changed after irradiation,as well as 102 points disappeared or increased;in neurula, 21 protein spots were expressed significant differences,and 63 points disappeared or appeared after irradiation.3) According to the results of the analysis of two-dimensional electrophoresis,16 spots of blastula and 25 spots of neurula were analysed by MALDI-TOF-MS.Altogether 11 spots were identified clear functions, including 7 kinds of proteins which expression reduced after irrdaition:67kD laminin receptor precursor,peroxidase 2,enolase 1,a similar homologous heat shock protein 70,MGC82602 protein,tRNA nucleotidyltransferase precursor and ribonucleoprotein Alb;and 4 kinds that decreased after irradiation:valosin containing protein,glycogen phosphorylase(brain),tubulin at 56D and Caspase-7.These proteins which were involved in stress response,cytoskeleton,inflammation,energy metabolism,protein synthesisa and degradation,cells proliferation and apoptosis expressed differentially before and after UVB irradiation significantly.4) The blastulas,gastrulas and the neurulas of Bufo bufo gargarizans were irradiated by 304μw/cm~2 UVB,then treated with immunohistochemistry staining.The results suggested that,after UVB irradiation,the expression of caspase-3 enhanced in animal pole of blastula but weakened in plant pole.In gastrula,the expression of caspase-3 increased in the number of positive cells.And in neural embryos,the expression of caspase-3 strengthens in the region of head,however weaken in venter.This basic research could help to reveal the organisms during the development of species evolution and variability,as well as genetic adaptation and the molecular mechanism of embryogenesis under the conditions of specific biological factors,and offered reference to environmental protection and health maintenance.