Prediction Of Pluripotent Factors And Its Expression During Skin Wound Repair In Sipunculus Nudus

Stem cell pluripotent factors are a kind of transcription factors that can induce mature somatic cells into pluripotent stem cells,which are very similar to embryonic stem cells.Since the discovery of pluripotent factors,more and more researchers have explored its links to the wound repair and regeneration capability.It has been suggested that pluripotent factors are the key point for cell reprogramming and also closely related to tissue regeneration.Moreover,to confirm the role of pluripotent factors in the cell regeneration is necessary to further describe it in different species,including simple and complex organisms.Thus,in this research the links between the cell pluripotent factors and skin wound repair were investigated in the marine worm Sipunculus nudus,a suitable candidate for its wound repair capability,comparable to higher organisms.A transcriptome sequence of S.nudus rebealed 17 types of stem cell pluripotent factors,including Sox2,c-Myc and Klf4 that are well known to play a key role during induction process in higher organisms.Also,the expression of Fthl in all tissues was much higher than the other stem cell pluripotent factors homeotic genes.On the other hand,the expression trends of stem cell pluripotent factors homeotic genes and regeneration homeotic genes in some tissues were similar.Based on the previous findings,4 factors(Sox2,Klf4,c-Myc,Fthl)and WNT signaling pathways(Wnt-7a and beta-Catenin)were selected to test the function during skin wound repair in S.nudus.In order to test the expression of the selected factors,the worms were cut longitudinally,10 mm length and 1 to 2 mm depth approximately,at a opposite to the anus by using a scalpel.Skin tissue from the incision was collected at 0 h,10 min,30 min,12 h,24 h,48 h and 72 h after cutting.Subsequently,qPCR and immunohistochemistry were used to test the expression of 6 stem cell pluripotent factors and the proliferation of cells during wound repair in S.nudus.Results showed that,all 6 factors reacted to the damage caused to the tissue,and the expression of Sox2,Klf4,Fthl,Wnt-7a and beta-Catenin were also up regulated during the process of skin wound repair in S.nudus.Moreover,reparation of the wound was observed 24 h after the injury.Conclusively,pluripotent factors showed apositive correlation with the skin wound repair in S.nudus.Same results have been obserbed in earthworms and newts,when the ability of regeneration is strong,the relationship between regeneration and pluripotent factors is high.