Effect of Annexin A3 on Dectin-1 Activation on Mouse Oral Tissue Organoids (MOTO).

Objectives: Dectin-1 is a type 2 transmembrane protein expressed in immune cells. The function of Dectin-1 has mainly been implicated in anti-fungal control by stimulating the phosphorylation of spleen tyrosine kinase (SYK). However, recent studies show that not only is the Dectin-1 receptor activated by fungal pathogen-associated molecular patterns, but also by endogenous cytosolic protein ligands such as Annexins found in apoptotic cells. The purpose of this study is to identify if Annexin A3 is associated with Dectin-1 activation as monitored by the phosphorylation of SYK in mouse oral tissue organoid (MOTO) cultures using a Western Blot.
Methods: Mouse tissues obtained from oral palatal and gingival and processed into MOTO samples. The study collected MOTO samples at intervals of Day 7, 10, 13, 17, and 20. They were treated with Beta-Glucan Peptides, and Annexin A3. Unstimulated MOTO cells were used as negative controls. Protein lysates of the cells were prepared for protein estimation and western blotting.
Results: Western Blot shows p-SYK and SYK expression in all samples, including the unstimulated cells. The addition of BGP or Annexin A3 did not induce further p-SYK, which indicates that the reagents did not activate the Dectin-1 receptor pathway.
Conclusions: This may be due to the loss of the Dectin receptor or the activation of alternative pathways in the MOTO cultures at those time points. Weak expression of SYK in MOTO after some time indicates further loss of SYK in these samples. However, based on the loading control lanes, there was equal levels of protein loaded across the board and was no protein degradation. Further experiments are required to verify the role of Annexin-A3 in association with the Dectin receptor and other potential molecular signaling molecules.