Physiologicallly relevant increase in temperature causes an increase in intestinal epithelial tight junction permeability

Karol Dokladny, Pope L. Moseley and Thomas Y. Ma
Am J Physiol Gastrointest Liver Physiol 290: 204-212,2006

http://ajpgi.physiology.org/cgi/content/full/290/2/G204

This paper discusses the impact of physiologically relevant increase in temperature (37 -41 degrees in Celsius) on epithelial tight junctions as well as the role of heat-shock proteins (HSPs) in regulating this heat stress. The study was done using a cell line known as caco-2 cells. Non-leaky epithelial tight junction barrier is "crucial in providing barrier function against epithelial penetration of pathogenic bacteria and toxic luminal antigens like endotoxins." As indicated by the study, the disruption of the tight junction barrier results in a "leaky" barrier that allows paracellular permeation of toxic luminal substances. The experimental data presented in the paper show that HSPs play an important protective role in preventing heat-induced disruption of the tight junction.

Western blot analysis was used to assess the HSP, occludin, ZO-1, and beta-actin protein that were expressed just as we did in lab. The researchers mixed the clear lysate derived from a centrifuged cell lysate with a Laemmli gel buffer, boiled for 7 minutes, and lated separated using SDS-PAGE gel. The protein from the gel was transfered to Nitrocellulose membrane overnight.

This paper was very interesting to me because it serves as a cautions to me as prospective tissue engineer to be mindful of little things that might end up ruining my engineered tissue. I was also intrigued by its mention as well as usage of certain techniques that I have learnt in class.