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Title: | 類感光細胞在雞松果體中的發育 The development of photoreceptor-like cells in chicken pineal gland |
Authors: | Chen-Ming Hao 郝貞明 |
Advisor: | 錢宗良 |
Keyword: | 松果體,松果腺細胞,視錐蛋白,類感光細胞, Pineal gland,pinealocytes,visinin,photoreceptor-like cell, |
Publication Year : | 2018 |
Degree: | 碩士 |
Abstract: | 本篇研究所欲探討對象雞是一種重要的神經發育模式動物,也是第一個被全基因定序的非哺乳類動物,牠填補了哺乳類與其他脊椎動物在演化上的空缺。於實驗室前人的研究中,已從雞的基因體中辨別並選殖雞的α-internexin同源基因,並確認基因序列、結構與其在雞胚發育過程中的表現情形。進而利用該基因序列設計製作雞α-internexin (chkINA)的多株抗體於後續實驗,從免疫組織染色獲得的結果顯示chkINA在雞的小腦區域與視網膜的發育過程中有較高表現,因此認為chkNA是神經元特定表現之中間絲蛋白,可應用於以雞為模式之發育研究中的神經細胞標記物。前人研究指出松果腺細胞 (pinealocytes) 在很多非哺乳類的脊椎動物中有類似視網膜感光細胞的功能。然而過去α-internexin在雞松果體中的表現尚未被探討,因此本研究主要目的在觀察雞松果體發育過程中,α-internexin在雞發育中松果體的類感光細胞的表現分布。本論文利用西方墨點法與免疫組織化學染色等方法研究α-internexin與視錐蛋(visinin) 在雞松果體蛋白質表現量與分布情形。chkINA的表現在雞松果體發育早期即被檢測到,而隨著松果體的發育而表現量減少。我們也發現chkINA在visinin免疫陽性的類感光細胞 (photoreceptor-like cells) 中也有一些表現。透過共軛焦顯微圖像證實chkINA和visinin共存於雞松果體中的類感光細胞中。chkINA不僅可以在視網膜發育期間出現在感光細胞中,而且也在雞松果體內的類感光細胞中有表現。此外,我們從超微結構中觀察,在青壯年雞松果體中可以發現類感光細胞有退化外節段 (outer segments)。這意味著青壯年雞松果體中的類感光細胞正逐漸失去光感官功能。綜合本研究結果,我們研究了chkINA在雞松果體類感光細胞中的表達和分佈模式。我們認為α-internexin未來可以應用成為研究雞松果體中類感光細胞的有用的特定標定物,並將可提供松果體中感光細胞演化發育生物學研究更多線索。 The animal model of this study is the chicken which is an important neurodevelopment model organism that links the evolutionary gap between mammals and other vertebrates. From previous studies, the mRNA sequence which encoding chicken neuronal intermediate filament α-internexin had been identified, also the gene structures and expression levels during developmental process had been characterized. The chicken α-internexin (chkINA) antibody was also generated via putative α-internexin sequences. The results from the immunohistochemistry showed that chkINA could be highly expressed during developmental process of chicken’s cerebellum and retina. Thus, chkINA might be a neuron-specific IF protein and could be a useful marker to be applied to neurobiological studies in chicken. It has been reported that some of pinealocytes are shown to have similar function to the photoreceptor of the retina in many non-mammal vertebrates. However, the expression of α-internexin in chicken pineal gland is still unknown. Hence, the purpose of this study is to investigate the expression pattern of chkINA in photoreceptor-like cells of the developing chicken pineal gland.The expression of chkINA and visinin in developing chicken pineal gland were examined by Western blot and immunohistochemistry. The expression of chkINA was detected in early stage of pineal gland and decreased following the development of pineal gland. We found that some expression of chkINA in photoreceptor-like cells were also Visinin immunopositive. chkINA and visinin co-exist in photoreceptor-like cells was demonstrated in the chicken pineal gland by confocal images. chkINA could be found not only in photoreceptors during retinal development but also found in the photoreceptor-like cells within chicken pineal gland. Besides, from our ultrastructural observations, the degenerating outer segments of photorecptor-like cell could be found in young adult chicken pineal gland. It could be implied that the photoreceptor-like cells is gradually losing sensory function of light in young adult chicken pineal gland. In summary, we have studied the expression and the distribution patterns of chkINA in photoreceptor-like cells of chicken pineal gland. We conclude that chkINA could be a useful photoreceptor-like cells marker to be applied for the study of chicken pineal gland. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70935 |
DOI: | 10.6342/NTU201801881 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 解剖學暨細胞生物學科所 |
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