Introduction

Tight junctions are multi protein complex, including occludin, claudin and JAMs, found
between neighboring epithelial cells that perform cell to cell adhesion and the regulation of the passage
of water, ions and other molecules through the epithelial membrane. Tight junctions are attcked by
bacteria that are know to cause symptoms of food poisoning through alternation of the cytoskeleton or
breaking down of important junction proteins. Escherichia coli, a bacteria associated with the caused
of food poisoning, plays a central role in decreasing transepithelial resistance (TER) and altering the
intestinal epithelial tight junction (TJ) structure(Shifflett et al.,2005). The bacteria release effector
proteins that breakdown junction proteins, leading to higher paracellular flux of molecules through the
epithelial membrane. The higher flux decrease transepithelial resistance, which is a measure of
electrical difference between the inside and outside of the epithelial membrane. The higher flux from
blood vessel through the epithelial membrane to the lumen of the intestine is the main cause of
diarrhea, a common symptom of food poisoning. Other bacteria can change the permeability of tight
junction through alternation of the cytoskeleton. Clostridium difficile plays a crucial role in
regulating the organization of the actin cytoskeleton(Nusrat et al., 2001), which lead to the higher
influx of water through the tight junctions. For this experiment, an unknown bacteria associated with
causing symptoms of food poisoning was tested to confirm its effect on tight junctions in the intestinal
epithelium. Looking at other food poisoning causing bacteria, I concluded that food poisoning bacteria
affect the tight junctions of epithelium in order to cause food poisoning, therefore the unknown bacteria
must interfere with the function of tight junctions in some way.

Experiment
For the experiment, the transepithelial resistance through an intestinal epithelial membrane will
be measured to determined the bacteria effect on the tight junctions. In epithelial cells with affected
tight junctions, the transpepithelial resistance decrease as a result of higher permeability of the

membrane. A probe is used to measure the voltage difference through the membrane. The epithelial
membrane used will composed of Caco-2, cell line derived from human colon adenocarcinoma, plated
in a monolayer fashion. Caco-2 was used for because it contains tight junctions similar to those found
in intestinal epithelium. The membrane will then be placed in an enclosed well that seperate it into two
halves. For the negative control of the experiment, the voltage difference of the membrane will be
taken before and after 12 hours in an incubator. The result should show no change in the transepithelial
resistance. As for the positive control, Escherichia coli will be added to the well in one compartment
and be placed in the incubator for 12 hours. Measurements will also be recorded before and after the
12 hours period, and a decrease in transepithelial resistance should be observed. Lastly, the unknown
bacteria will be placed in the well similar to the positive control. If a decrease in the transepithelial
resistance is measured, it means the unknown bacteria caused food poisoning through alternation of the
tight junctions. However, if no changes is detected in the resistance, it means the hypothesis is wrong
and that the mechanism of this unknown bacteria should be examined closely.