Research projects

To be able to analyse the contribution of CRB2 protein to the development of pigmentary epithelial bonds and its possible involvement in retinal dystrophies, one of the first objectives is to show that CRB2 protein is present and where it is located, and/or the other two members of the CRB family of proteins in this cell type. To this end, and to date, using molecular, biochemical and morphological techniques, CRB2 as well as CRB3 have been shown to be present in the pigmentary epithelium; while excluding the possibility that CRB1 is present too. Until now, these studies were the first to demonstrate these facts. Morphological analyses show that CRB2 is located very precisely in the apex and sides of the pigmentary epithelium, within the area of adherent bonds. On the other hand, to be able to analyse the modifications of the adherent bonds in the pigmentary epithelium under different conditions of in vitro CRB2 expression, a technique was perfected to obtain primary cultures of pigmentary epithelium obtained from the eyeballs of mice. Thus, after 5 days of culture, few cells were observed with fusiform morphology and the characteristics of proliferative cells. After several days of culture, many of these cells presented a morphology typical of pigmentary epithelial cells, because they had started to synthesise proteins that are characteristic of this cell type and presented pigment. Finally, among other experiments, the transepithelial resistance of the cultures is being measured, making it possible to determine the strength of the bonds formed between the cells. It was found that this transepithelial resistance does in fact increase during the time that the cells are kept in the culture. This technique will make it possible to achieve one of the aims of this project, which is to check whether changes occur in the strength of the adherent bonds of the pigmentary epithelium under different conditions of CRB2 expression, together with whether these changes affect the functions of this type of cell in the retina.