The use of transgenic and knock-out mice in the investigation of ocular surface cell biology.

The transgenic and knock-out mice created by transgenesis and gene targeting techniques are very useful for elucidating the pathophysiology of human diseases caused by altered genetic functions. Many of the experimental mouse lines exhibit ocular surface disorders. However, embryonic lethality and congenital defects found in many of the transgenic and knock-out mice preclude their use for studying the consequences of altered genetic functions in adult animals. To circumvent these difficulties, we have established binary inducible mouse models, using the corneal keratocyte-specific keratocan promoter, and the tetracycline-inducible gene expression system (reverse tetracycline transcription activator--rtTA). In these models, the animals function normally until they are fed doxycycline, thus inducing the overexpression of inserted transgenes by keratocytes. We have also developed inserted rtTA and Cre reporter gene constructs to create genetically modified mouse lines that have tissue-specific gene alterations to study acquired conditions, e.g., wound healing and irregular hormone and cytokine signaling that offsets homeostasis in adults. Furthermore, the genes that are ubiquitously expressed in many tissues can be specifically ablated solely in ocular surface tissues to examine their function, since the loss of such a gene in ocular surface tissues will not be life-threatening. It is noteworthy that these altered mouse lines can also be used as models for the development of therapeutic treatment regimens of diseases using gene therapy and stem cell strategies.