RETINAL GENE TRANSCRIPTIONAL REGULATION AND NEUROPROTECTION OF PHOTORECEPTORS

GENE TRANSCRIPTIONAL REGULATION;RETINA;SYNAPTOTAGMIN 11;S-ANTIGEN;PHOTORECEPTOR NEUROPROTECTION;not listed

Shaw, Gillian CSTEINER, JOSEPH ;

Johns Hopkins University

关键词: GENE TRANSCRIPTIONAL REGULATION;    RETINA;    SYNAPTOTAGMIN 11;    S-ANTIGEN;    PHOTORECEPTOR NEUROPROTECTION;    not listed;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/60507/SHAW-DISSERTATION-2015.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

My PhD research included two distinct projects. Project I examined the transcriptional regulation of two genes expressed in the retina, synaptotagmin 11 (Syt11, retinal ganglion cell-enriched) and S-antigen (Sag, photoreceptor-enriched). For each, a cloned fragment was identified that showed promoter activity in its respective cell population. A deletion analysis and subsequent bioinformatic analysis were used to predict transcription factors (TF) binding within the region of interest, and then binding site mutant promoter-reporter constructs were generated and tested. Finally, siRNA knockdown of TFs of interest was followed by endogenous expression analysis. I identified regions of interest for each gene (Syt11 -192 to -41 bp and Sag -279 to -182 bp) but knockdown of multiple TFs in each cell population yielded inconclusive results.In Project II, I evaluated sunitinib, a multiple kinase inhibitor that is neuroprotective for retinal ganglion cells, for its ability to promote photoreceptor (PR) survival in mouse models of PR degeneration. Based on the findings of a collaborator who found that sunitinib promoted the survival of PRs in a rat model of an autosomal dominant retinal degeneration (Rho S334ter), I studied the retinal expression of dual leucine kinase (DLK) protein (known to be a target of sunitinib) during retinal degeneration (RD). I observed an increase in DLK abundance in degenerating retinas, which persisted after most PRs had degenerated, suggesting that DLK accumulation occurs, at least in part, in a cell type other than PRs. I tested the effect of sunitinib on photoreceptor survival in the Rho Q344ter and the rd1 (Pde6b mutant) mouse RD models, but under the conditions tested did not observe evidence of increased PR survival. Additionally, I tested the ability of genetic deletion of Parp1 to promote PR survival in the sodium iodate and rd1 mouse models of RD. Parp1 null PRs did not show evidence of increased survival at 36 days post sodium iodate administration. Several litters of Parp1 KO/rd1 animals had increased PR survival. Further studies are needed to more fully define the potential role of sunitinib, DLK inhibition, and PARP1 inhibition in photoreceptor neuroprotection.