Modulation in proteolytic activity is identified as a hallmark of exogen by transcriptional profiling of hair follicles.

Exogen is the process by which the hair follicle actively sheds its club fiber from the follicle. However, little is known about signals that govern the cellular mechanisms of shedding. Here, we have identified factors that are important in regulating either the retention or release of the hair club fiber from its epithelial silo within the follicle. Using the vibrissa follicle as our model, we isolated follicle segments containing club fibers and surrounding follicle tissue at different time points before their natural release from the hair follicle. We then performed microarray analysis to identify key molecular changes as the club fiber approached final release. Among the different classes of genes that were identified, we found changes in the expression pattern of protease inhibitors and proteases, suggesting that proteolysis may mediate fiber release, either through terminal differentiation or proteolytic cleavage. We validated transcriptional changes using reverse transcription-PCR, and further immunofluorescence analysis indicated that protease inhibitors surrounding the club fiber may have an important role in regulating the process of club fiber shedding. Our findings also highlighted that molecular differentiation of the innermost layer of cells immediately surrounding the club fiber, the companion(CL), is likely to be important in hair shedding.