Thymosin beta 4 dynamics during chicken enteroid development

by Acharya, Mohan; Liyanage, Rohana; Gupta, Anamika; Arsi, Komala; Donoghue, Ann M.; Lay, Jackson O., Jr.; Rath, Narayan C.

The sheared avian intestinal villus-crypts exhibit high tendency to self-repair and develop enteroids in culture. Presuming that this transition process involves differential biomolecular changes, we employed matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) to find whether there were differences in the spectral profiles of sheared villi versus the enteroids, assessed in the mass range of 2-18 kDa. The results showed substantial differences in the intensities of the spectral peaks, one particularly corresponding to the mass of 4963 Da, which was significantly low in the sheared villus-crypts compared with the enteroids. Based on our previous results with other avian tissues and further molecular characterization by LC-ESI-IT-TOF-MS, and multiple reaction monitoring (MRM), the peak was identified to be thymosin beta 4 (T beta 4), a ubiquitously occurring regulatory peptide implicated in wound healing process. The identity of the peptide was further confirmed by immunohistochemistry which showed it to be present in a very low levels in the sheared villi but replete in the enteroids. Since T beta 4 sequesters G-actin preventing its polymerization to F-actin, we compared the changes in F-actin by its immunohistochemical localization that showed no significant differences between the sheared villi and enteroids. We propose that depletion of T beta 4 likely precedes villous reparation process. The possible mechanism for the differences in T beta 4 profile in relation to the healing of the villus-crypts to developing enteroids is discussed.

Molecular and Cellular Biochemistry
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