Identification and functional analysis of zebrafish OPTN

• Main Authors: Ying-Li Wang, 王映酈
• Other Authors: 楊建洲
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/7mq3tj

碩士 === 中山醫學大學 === 生物醫學科學學系碩士班 === 106 === Glaucoma is a leading cause of irreversible visual impairment and blindness resulting from progressive degeneration of retinal ganglion cells (RGCs) and optic neuropathy. Optineurin was originally identified as a gene responsible for primary open-angle glaucoma. The gene, OPTN, codes for the protein optineurin, which is involved in a variety of functions including regulation of endocytic trafficking, autophagy, immune response, mitosis and NF-κB signal transduction. Many missense mutations in optineurin have been reported and the association of the mutation with the disease varies in different populations. Optineurin is localized to pathological structures also seen in several neurodegenerative diseases such as amyotrophic lateral sclerosis, Alzheimer''s disease, Parkinson''s disease, etc. However, functional alterations caused by mutations in optineurin are poorly understood. In this study, we investigated the physiological functions of OPTN in the zebrafish embryonic development. RT-PCR analysis of zebrafish OPTN transcripts were revealed significantly in the 120hpf embryos. Meanwhile, whole-mount in situ hybridization (WISH) signals were also observed in the 120hpf embryos. In addition, we established the fusion protein construct OPTN-TagRFP that was injected into embryos in the presence or absence of OPTN-targeting morpholinos. We have found that OPTN- targeting MO was able to specifically knockdown the expression of TagRFP-tagged OPTN fusion proteins in a dose-dependent manner. These data may be useful for studying early POAG pathophysiology.