Preliminary Screening Of Differentially Expressed Genes Between Male And Female Reindeer Antler Reserve Mesenchyme

Deer antlers are the only mammalian appendages capable of regeneration, and thus provide a unique model for researching the mechanisms of mammalian organ regeneration. Antler growth center cell’s growth rate of proliferation is more than 30 times as fast as cancer cell, And did not find any signs of cancer. Understand the molecular mechanisms can provide new goals and direction of cancer research. Reindeer are the only deer species in both male and female horned. However, there are large differences in their male antler, and have different antler shedding time. The study of differentially expressed genes of male and female reindeer antler may be easier to figure out antler regeneration and growth of molecular mechanisms.In this experiment, we applied the reserve mesenchyme of antler for the study. The total RNA were extracted by Trizol method, using differential display reverse transcription PCR screen transcripts differentially expressed during male and female reindeer antler. And then polyacrylamide gel electrophoresis、secondary amplification agarose gel recovery、product cloning and sequencing to detect false positive of differ -ences fragment. At last, we used real-time PCR to identify the results. DDRT-PCR reaction was performed by using 30 primers combinations of 3 anchored primers coupled(A、B、C) with 10 random primers(a、b、c、 d、e、f、g、h、i、j). The product did SDS-PAGE and silver staining.52 expressed sequence tags were gained and 7 were recovered successfully. The sequences were compared in NCBI’s BLASTN.5(Ae1、Af1、Af3、Ce1、Ci2) are known genes, the 5 sequences are between 100-400bp. Ael shows high homology (99%) with bovini TPM4, TPM involve in the regulation of muscle contraction, cell motility and other life activities, also reported that the expression of tropomyosin down was related with the occurrence of cancer; Afl shows high homology (94%) with bovini SMAD5. SMAD is related with TGF-(3 Signal Transduction. SMAD5 promote osteogenic effect; AD shows high homology (94%) with bovini HMGB1. HMGB1 can stabilize the nucleus and assist the transcri-ptttion. When it secretes into the extracellular, it may occur with cancer-related; Cel shows high homology (98%) with Tibetan antelope COL6A3. COL6 is the main component of bone and muscle; Ci2 shows high homology (92%) with sheep SLC17A5. SLC17A5 expression Sialin transport Sialic acid in the cells. In this paper, preliminary finding:Ael and Af3 of the male reindeer antler reserve mesenchyme are higher expression levels than the female. Af1、Ce1、Ci2 of the female reindeer antler reserve mesenchyme are higher expression levels than the male. The result of COL6A3 by real-time PCR Consistent with the DDRT-PCR, but the differentially expressed is not significant(P>0.05).