Correlating Proteins Isolation And Identification Using Two-Dimensiona Electrophoresis And Mass Spectrometry In OPIDN Induced By TOCP In The Cerebrum Of Hens

Objective:To screen the proteins of differential expression in the cerebrum of hens in OPIDN induced by TOCP using Two-Dimensional electrophoresis and Mass Spectrometry to "TOCP and PMSF dual receptors influence" hypothesis as the theoretical basis, and to explore the mechanism of TOCP in OPIDN.Methods:Twenty four roman hens,10 months old, weighing 1.5-2. 0kg,were randomly divided to four groups, and six hens in one group. Adaptive feeding animals a week, according to the "double receptor of hypothesis," these four groups were designated as the control group, given by administration once 1000mg/kg TOCP group 24 hours after subcutaneous injection of 40mg/kg PMSF through a grant administered 1000mg/kg TOCP group, subcutaneous 40mg/kg PMSF group. Seven days after exposure the animals were sacrificed, Their cerebrum were dissected and homogenized in ice bath. Total proteins of the cerebrum tissues were separated by isoelectric focusing as the first dimension and SDS-PAGE as the second dimension. The 2-DE maps were visualized after silver staining and analyzed by ImageMaster2D software. In the two-dimensional electro-phoresis expression profiles on select point number 1612,1171,1457,1067 filtered and used mass spectrometry for the identification. Finally get the results of protein identificationResults:In the total proteins from the control group and the experiment group,1332 and 1295 protein spots were detected respectively. Comparison analysis showed that 78.31% of the total protein spots were matched between the two groups Compared with control group,262 discrepancy protein spots were detected in experiment group and 32 protein spots of 4 fold change. Mass spectrometry results for the 1612 points, mainly glutamine synthetase, and isoelectric point of 8.26, a molecular weight of 31.66 kD.1171 points for the heat shock protein 70, and isoelectric point of 7.57, a molecular weight of 41.81 kD.1457 points for the L-lactate dehydrogenase B, and isoelectric point of 7.71, a molecular weight of 35.58 kD.1067 points similar to homer-lb, isoelectric point of 5.54, a molecular weight of 44.40 kD. And these four proteins and the control group, TOCP groups were down regulated (P< 0.05); the other two groups and the control group no significant changes in expression.Conclusion:Through "organic phosphorus compounds and PMSF double Receptor Hypothesis" from TOCP exposure group were screened out of 236 protein spots may be associated with delayed neurotoxicity, which homer-1b, glutamine, glutaminase three protein may occur during OPIDN key protein.