The childhood eye cancer retinoblastoma originates from the retina and its development is initiated while the foetus is in the uterus. Retinoblastoma has a reported incidence of 1 in 15-18 000 live births, and approximately 90% of all patients are diagnosed before the age of 5. The occurrence of retinoblastoma is usually detected by the parents and the most frequent symptoms are leukocoria (white pupillary reflex), strabismus (squinting) or if the child complains of visual problems. Retinoblastoma is diagnosed by examination under anaesthesia and documentation by RetCam. It is treated with various cytostatic agents, or by laser. If the treatment is unsuccessful, or there is a risk that the tumour cells will spread and form metastases, the eye is removed. Previous studies have indicated that the cell type from which the tumour arises, the cell-of-origin, may be the cone photoreceptors and/or their immediate interneuron, the horizontal cells. Determining the cell-of-origin for retinoblastoma is an important goal, however, understanding the molecular mechanisms that distinguish the photoreceptors and the horizontal cells from the other retinal cells may prove just as important for understanding this disease. The aim of my project has been to develop, optimise and validate methods to label, map and target expression to photoreceptors and horizontal cells in the chicken embryonic retina. We have successfully established several methods that test the expression pattern of conserved, regulatory DNA sequences, and have performed short- and long-term expression of various genes that have been reported to be involved in cell cycle regulation and cell fate determination. One of my most important findings was that a region from the RXRγ gene allowed us to specifically target the photoreceptors and horizontal cells. Our previous knowledge, together with the newly established tools, puts us an important step closer towards understanding the development and behaviour of the retinal photoreceptors and horizontal cells, however, further studies are of course needed.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2006. / Härtill 4 uppsatser.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 91-101).

Previous studies from our lab have shown that the Activating Protein- 2 (AP-2) transcription factors, AP-2α and AP-2β, are important in retinal development. It was discovered that these are co-expressed in developing horizontal cells and postmitotic amacrine cells. To understand their role in retinogenesis, and the impact of their deletion on the adult retina, a double mutant mouse model was created, AP-2αKI/flox/AP-2β-/flox. The neural retina of the AP-2αKI/flox/AP-2β-/flox mice was examined in the current study using histological, immunofluorescent and electron microscopy (EM) techniques at embryonic, post-natal and adult stages. These double mutants displayed a variety of abnormalities in the inner retina. Loss of AP-2α and AP-2β at E10.5 led to a complete absence of developing and mature horizontal cells. This loss was associated with changes in the outer plexiform layer, which diminished from two to four months of age. There were also defects with photoreceptor ribbons in which triad synapses failed to form, and instead led to rudimentary, spherical-shaped ribbons. There was also significant retraction of photoreceptor axons. Furthermore, this study was able to infer a role of AP-2α and AP-2β as acting upstream of the Onecut-1 protein, which targets Lim1 and Prox1 to direct horizontal cell genesis. Examining amacrine cells of the double mutants shows evidence that AP-2α and AP-2β are involved in the mosaic arrangement pattern of amacrine cell bodies and axons. Previous work on embryonic double mutants displayed clustering of amacrine cells. This study observed abnormalities in the dendrites of the inner plexiform layer, which consists of amacrine cell processes. Taken together, the work presented in this thesis implicates the redundant requirement of both AP-2α and AP-2β in development of horizontal cells and patterning of amacrine cells in the neural retina. / Thesis / Master of Health Sciences (MSc)