Date of Award
Honors College Thesis
Biological Sciences BS
Hao Xu, Ph.D.
Jake Schaefer , Ph.D.
Sabine Heinhorst, Ph.D
Mast cells secrete a variety of biological mediators crucial for innate and adaptive immunity. One such mediator is Tumor Necrosis Factor (TNF). TNF released from mast cells and other immune cells has been shown to elicit complex signaling pathways in target cells, resulting in the production of pro-inflammatory cytokines and/or immune cell recruitment. At the molecular level, TNF secretion/exocytosis is mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) and other regulatory proteins that work together to promote the fusion between secretory granules and the plasma membrane. One regulatory protein, Munc13-4, which facilitates the priming of secretory granules, was recently shown to be partially responsible for TNF release from mast cells. This discovery suggests that another Munc13 homolog (BAIAP3 or Munc13-1) could be involved in TNF secretion. In this study, I cloned BAIAP3 cDNA into a lentiviral expression vector, upstream of an EmGFP tag. Following sequence verification, the newly engineered pLVX-IB-BAIAP3-EmGFP expression construct was introduced into RBL-2H3 mast cells along with pCDH-TNF-SBP-mCherry. Confocal microscopy showed that BAIAP3 is partially co-localized with TNF in granular structures, supporting the notion that BAIAP3 might be the missing Munc13 in TNF release. This new construct will be used in the future to rescue any secretory defects in BAIAP3 knockout cells.
Keywords: Mast cells, mediators, TNF, SNARE, BAIAP3, secretion, construct
Copyright for this thesis is owned by the author. It may be freely accessed by all users. However, any reuse or reproduction not covered by the exceptions of the Fair Use or Educational Use clauses of U.S. Copyright Law or without permission of the copyright holder may be a violation of federal law. Contact the administrator if you have additional questions.
Little, Ireland Ann, "Generation of BAIAP3 Expression Construct for Subcellular Localization and Phenotypic Rescue" (2022). Honors Theses. 869.
Available for download on Saturday, August 10, 2024