Differential Proteome Analysis Of The Fetal Heart Tissue In The Embryo Different Developmental Stage

Objective The development of heart involve intricacy process. A number of proteins were involved in the process. In the present study, we analyed the proteome of fetal heart tissues from Day 14d, 16d and 18d in NIH mice. Specially, we studied differential protrome of fetal heart of mice on 14d, 16d and 18d of pregnancy by using comparative proteomics. The aim of this work is to acquire knowledge about the mechanism in the development of heart. Methods 1. Found the NIH mouse pregnant animal model, the fetal heart tissue of 14d, 16d and 18d were separated from embryo of NIH mice, then immediately stored in -80℃ untill processed. 2. The proteins of pregnant 14d, 16d and 18d were separated by two-dimensional gel electrophoresis. The two-dimensional gels were stained by Coomassie Blue and subjeted to analyse by PDQuest software. 3. Differential protein spots were excised from the gels and digested with trypsin. The peptides were analysed by MALDI-TOF-MS, the peptides mass fingerprint(PMF) was obtained. PMF were analysed by Moscot software. Results 1. 2-D PAGE profile showed that the proteins in the fetal heart were located mostly in the range of mass 20-116 kda and isoelectric points(pI) at 4.0-8.5. Analysing from the profile by PDQuest software, there are 565 protein spots in pregnant 14d, 711 protein spots in pregnant 16d, 773 protein spots in pregnant 18d. Comparing between pregnant 14d and 16d, at least 22 protein spots were up-regulated by over two fold and 8 protein spots were down-regulated. Comparing between pregnant 14d and 18d, at least 39 protein spots were up-regulated and 11 protein spots were down-regulated by over two fold respectively. 2. PMF of 14 differential protein spots were searched by database. 4 protein spots of them were identified unambiguously. They are Heat shock protein 60 (Hsp60), cytoskeletal 6G, mitogen-activated protein kinase kinase kinase4(MAPKKK4 or MEKK4). Among them , another 2 protein spots were identified as MAPKKK, but they were not isoform. Conclusions 1. during the development of fetal heart of mice, a lot of proteins were participated in this process. 2. Molecular weight of Hsps 60KDa. It exist mainly in cytoplasm and mitochondria and play a role of molecular chaperonin, viz. protein folding, transportation orientation. When the body is stressed, the synthesis of Hsp60 can be markedly induced to stabilize and refold proteins. Hsp60 is benefited for the function of heart as it is expressed in the different time of embryo. 3. MAPKKK4 or MEKK4 is one of the members of the major systems participating in the cellular signal transduction of MAPK. It can activate signal pathway of JNK and regulate consequently cell division and apoptosis. By a series of signal transduction, it play an important role for cell growth, differentiation, development and death. 4. Cytoskeleted 6G participate in the forming of hair. It is possibly pollution of protein.