Detection And Analysis On Telomerase And Development Related Proteins In Cells Of Deer Antler

Telomerase is a special reverse transcriptase, which also has a telomerase RN A (TR) component, and reverse transcriptase (TERT) component..Telomerase is able to utilizes TR as the template extension telomere. replace lost telomeric sequences preventing the cells to enter senescence, so that the cells continued to divide. it is power source of a cell can be immortalized. In most normal somatic cells. activity of telomerase was significantly inhibited, and in about 85% of the tumor cells. stem cells and reproductive cells can be detected in the high activity of telomerase.Deer antlers are the only mammalian organ capable of repeated rounds of regeneration; every year they are seasonal shed and regrow,in fast growing period, it can exceed 1-2 cm per day, representing the fastest rate of organ growth in the animal kingdom.Most of researches suggested that the rapid growth of deer antler depends on high-speed proliferation of cell in proliferative zone at the tip of antler. Antler development is a uniquely intense and complex process.although has been discovered IGF-1, PEDF, PTHrP and Wnt signaling pathways involved regulation antler growth and development, However, the underlying moleculer mechanism remains poorly understood. still only at the initial stage of the study.Telomerase as a kind of reverse trabscriptase which can maintained rapid cell growth.whether playing a role in grow and development of antler.has not been reported.In present research, the updated TRAP argentum-dying method and Bandscan software was used to detect and analyzed the activity of telomerase in mesenchyme. precartilage. cartilage and sclerotic tissue of antler tip during the growth stage. The result showed that the activity of telomerase was detected in the mesenchvme. precartilage and cartilage but not in the sclerotic tissue. The relative telomerase activity in the mesenchyme, precartilage and cartilage was 91.2.46.4 and 13.7. respectively.Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of telomerase reverse transcriptase (TERT) in these sample. The result showed that the relative expression pattern of TERT was similar to relative activity pattern, verify the reliability of the results of TRAP.Longitudinal growth of antler is by a process of modified endochondral ossification.the cell in mesenchymal layer of proliferative zone is the stem cell reserve layer. Mesenchymal cells is rapid proliferation and continue to differentiate into in various differentiation degrees cell of cartilage cells was led to the rapid growth of velvet. In order to further explore the molecular mechanism of antler growth and development,2D-DIGE combind mass spectrometry analysis was used to comparison and analysis mesenchymal layer tissue and percartilage layer tissue differences inprotein expression in order to discovery new antler growth associated protein.Intensity changes of 28 spots were detected with statistical significance. And 12 of them were identified by MALDI-TOF/TOF successfully.12 protein spots of themup-regulated include F-box protein 22,GATA-6,ZRANB2.NADH dehydrogenase subunit, Niemann-Pick type Cl. L-gulono-gamma-lactone oxidase otherwise 5 down-regulated include CD38 antigen.collagen typeⅢalpha 1.β-tubulin.cystic fibrosis transmembrane conductance regulator and hornerinThe analysis show that express a variety of signaling molecules, pro-apoptotic protein, promoting cell growth factor and metabolic enzymes in mesenchymal layer. GATA-6. F-box22 and splicing protein ZRANB2 is the firstly been detected in antler, they may play an important role in regulation growth and development of antlerThis study was the first time confirmed that the telomerase was expression in antler tip during the growth stage. the relative telomerase activityt is different among issue layers of antler tip. and with the increase of cell differentiation. the telomerase activity will be decreased. indicating that telomerase may be involved in cell proliferation and differentiation of antler. telomerase is one of reasens for rapid growth of antler.The first time using 2D-DIGE technology to analysis mesenchymal layer which key parts of antler growth and development,and found some protein which have not been reported was expression in mesenchymal layer.they may play an important role in antler growth and development process.