Faim Expression From Stress In Mice Spleens
INTRODUCTION: Fas Apoptosis Inhibitory Molecule (FAIM) is a protein that has been implicated as having an important role in cellular stress responses and acts to protect the cell from apoptosis.
OBJECTIVE: Given its role in opposing cellular stressors, we sought to determine whether the level of FAIM is constitutively present or responds to stress.
METHODS: We examined primary cells, placed under conditions of oxidative or heat stress. Cells were obtained from the spleens of two types of mice: a wild-type (WT) mouse, and a heterozygote mouse in which green fluorescent protein (GFP) is knocked-in to the FAIM locus. Single cell suspensions of splenocytes were exposed to oxidative stressors including hydrogen peroxide and menadione, or heat stress of 40℃ in a water bath. The exposures were provided for varying periods of time, and the total production of GFP was assessed using flow cytometry as an indication of FAIM locus activation, with WT mice acting as the negative control.
RESULTS: Exposure to high levels of oxidative and heat stress led to increased mean GFP expression in the cells for the next 48 hours by 35%. Complicating matters was a decreased survivability in the cells exposed to higher amounts of oxidative and heat stress.
DISCUSSION: These results indicate that FAIM does respond to oxidative and heat stress with upregulation. Future efforts will involve replicating these experiments with cell lines that are much less susceptible to impairment of viability.