Screening Of Differential Proteins In Different Parts Of Pedicle Periosteum Of Sika Deer

Antler is the unique organ which can fully regenerate in the mammals, and it can be used as the superexcellent biomedical model to explore the regeneration process of mammalian tissues or organs. Further studies showed that deer antler regeneration is taking place from pedicle periosteum, and it is based on stem cell regeneration process, which depends on the activation of the stem cell on the distal of pedicle periosteum periodicity. Most of the former studies on antler regeneration are morphological or histological or genic research, while little studies on proteomics. Therefore, this research used deer anlter pedicle periosteum as the experimental material, and two-dimensional electrophoresis (2-DE) and comparative proteomic and bioinformatics technology were used to research the different expression proteins in different parts of pedicle periosteum. This research will lay the theoretical foundation for the explaination of deer antler regeneration.1,we tried to find out the best methods of smashing and extracting pedicle periosteum proteins, parameters on two-dimensional electrophoresis of pedicle periosteum proteins, and then we established two-dimensional electrophoresis system which was suitable for analyzing pedicle periosteum proteins. Results were as follows:(1) Cutted the periosteum tissue into fragments at 2mm×2mm, then precoolled them with liquid nitrogen, and ground when liquid nitrogen overlapping them. It came out that only in this way, can we extract pedicle periosteum tissue sufficiently.(2) The lysate ingredient contained 7M urea, 2M thiocarbamide, 4% CHAPS, 65mM DTT, 1mM PMSF and 0.2% Bio-lyte, this method was propitious to the sample preparation of pedicle periosteum proteins which were analyzed by two-dimensional electrophoresis.2,we made comparative analysis on protein contents from different parts of pedicle periosteum by using two-dimensional electrophoresis system which had been established above, then we detected different protein spots, verificated and classified them with mass spectrometry and bioinformatics technology. The results were as follows:(1) In the 2-DE map of different parts of pedicle periosteum, the number of the upper protein spot was more than the lower, which were 269, 311 and 241, 241, respectively, and the matching rate of different parts was up to about 50%.(2) The results of preliminary mass spectrometry on protein spots showed that there were 47 proteins, which can be divided into two categories. One classification was active protein, including 42 proteins, which accounted for 89.36% of identified proteins. The other one was non-active protein, containing 5 proteins, which was 10.64% of identified proteins. Among them, there were five protein sorts in active protein according to the different functions, and they were 14 catalytic enzymes, which was 29.79% of identified proteins; 13 proteins controlling growth and differentiation, which accounted for 27.66% of identified proteins; 8 transport and storage proteins, which took up 17.02% of identified proteins; 3 movement proteins, which covered an proportionality of 6.38% during identified proteins; 4 defense proteins, which made up 8.51% of identified proteins.3,At last, On the functional analysis of identification proteins we found six proteins: PKM2,MAPK1,pigment epithelium-derived factor,peroxiredoxin 4,heat shock 90kDa protein 1, alpha,fibulin 5, which closely related with antler regeneration and might be participated in signals transduction and molecular regulation of antler regenerate. In those proteins, pigment epithelium-derived factor and fibulin 5 have the function of inhibiting angiogenesis. In the process of antler regenerate which started by pedicle periosteum, regulated the expression of those two proteins might be one of the ways which regulated and controled the rapid growth of antler to avoid carcinogenesis.