Expression Analysis Of Tenascin-R(TNR) In The Regeneration Process Of Sea Cucumber Intestine

Under unfavorable environment,sea cucumber Apostichopus japonicas will discharge all or part of the internal organs to defend.This kind of defense means evisceration.When the environment is suitable,sea cucumber can regenerate the whole internal organs.The process is accompanied by changes and reconstruction of extracellular matrix components.As an important component of ECM,Tenascin-R(TNR)plays an important role in the development and regeneration of the nervous system of higher animals.It also participates in the division and differentiation of other types of cells.In this experiment,we obtained the c DNA fragment of the Tenascin-R gene and the3'-UTR of a total of 1524 bp,including an open reading frame of 1194 bp,encoding 397 amino acids.The protein has a molecular weight of approximately 45.1 k Da and contains two domains,EGF and FBG.It is a hydrophilic protein with no transmembrane domain.It has an isoelectric point of 5.14 and contains 64 phosphorylation sites.Induction of Aj-TNR protein with p GEX vector,the expression results showed that the protein was a soluble protein.Real-time PCR results showed that m RNA expression showed a trend of increasing first and then decreasing,reaching the maximum on the sixth day.Western blot results showed that the expression trend is similar to m RNA expression trend.By immunofluorescence assay,it was determined that TNR protein was widely expressed in various tissues and concentrated in the muscle layer of the regenerated intestinal tissues.Analyzing TNR expression patterns,we speculate that TNR may be an important boundary for intestinal nerve regeneration,which becomes a key point in the regeneration process and controls the growth of nerve tissue after vomiting.This study analyzed the expression pattern of TNR during the regeneration of Apostichopus japonicas,and the results showed that TNR may play a role in the intestinal regeneration of Apostichopus japonicas.