請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/30357
標題: Cloning and expression of a soluble guanylyl cyclase gene in the brain of honeybee, Apis mellifera
蜜蜂腦部水溶性鳥苷酸環化酶之選殖與表現
作者: 徐婷芸
Hsu, Ting-yun
關鍵字: 水溶性鳥苷酸環化酶
一氧化氮
蜜蜂
出版社: 昆蟲學系
摘要: Soluble guanylyl cyclases (sGCs) are an important target for endogenous nitric oxide (NO). sGC are heterodimeric enzymes consisting of α and β subunits, and have been considered as a key factor in the regulation of synthesis of cyclic GMP, a second messenger of signal transduction. The aim of this study is to discover the possible function of sGC in honeybee brain, and to investigate the gene at different ages. The first β1 subunit gene (Amgcβ1) was cloned from the honeybee brain. The Amgcβ1 consisting of 2010 bp contains an open reading frame and encodes a protein of 603 amino acids with a predicted molecular weight of 66.33 kDa. The sequence exhibited 76.7 % amino acid identity with the tabacco hornworm, Manduca sexta β1 amino acid sequence, 57.2% with malaria mosquito, Anophele gambiae, 55.7% with fruit fly, Drosophila melanogaster. RT-PCR analysis demonstrated that the transcripts was mainly present in head and indicated that Amgcβ1 transcript was present at a variable levels in any stages of development of adult workers. Thus, RT-PCR analysis that treated with 100 μg of JHA show no significantly different from the normal worker brain after 24 and 48 hours of the treatment. In situ hybridization analyses show that Amgcβ1 was expressed at high level in the laminar monopolar cells, lamina, outer chiasma and medulla of optic lobe, the soma of Kenyon cells in the mushroom body and the glomeruli of antennal lobe. sGC was shown in the visual and olfactory systems using in situ hybridization for the Amgcβ1, suggesting that cGMP-dependent signaling pathway may play an important role in the visual and olfactory systems of honeybee.
水溶性鳥苷酸環化酶 (soluble guanylyl cyclase; sGC) 在NO/cGMP訊息傳遞中受到一氧化氮 (nitric oxide, NO) 的活化,進而將GTP轉變成cGMP。本研究從蜜蜂Apis mellifera L.工蜂的腦部選殖得一β次單元的基因,並將之命名為Amgcβ1。Amgcβ1全長為2010 bp,具有大小為1809 bp之開放讀架 (open reading frame),序列末端具有23個腺嘌呤 (A) 所組成之poly (A) tail,5’-及3’-端分別具有94及106 bp之未轉譯序列 (untranslation sequence)。Amgcβ1由603個胺基酸所組成,預估分子量為66.33 kDa,經序列比對後發現Amgcβ1與已發表之昆蟲sGC β次單元基因最為相近,與菸草天蛾 (Manduca sexta) 的MsGC-β1有76.7%的一致性,與甘比亞瘧蚊 (Anopheles gambiae) 的GCSβ一致性有57.2%,而與黃果蠅 (Drosophila melanogaster) 的Dgcβ1有55.7%的一致性。RT- PCR的分析結果顯示,Amgcβ1在頭、胸及腹均有表現,但頭部表現量較高。Amgcβ1在羽化後第0天即有表現,但在羽化後第7天及第14天之工蜂表現量則降低,然後在羽化後第21天及第28天之工蜂表現量則再度上升。對初羽化工蜂處理100 μg青春激素異構物(juvenile hormone analogue, JHA)-百利普芬 (pyriproxyhen),在處理後的24及48小時內 Amgcβ1的表現沒有明顯變化。原位雜合結果顯示Amgcβ1表現在視葉 (optic lobes) 的lamina monopolar cells、lamina、outer chiasma及medulla,嗅葉的glomeruli、soma,以及蕈狀體 (mushroom bodies) 的Kenyon cells等組織,推測Amgcβ1在視覺及嗅覺的形成,以及經視覺產生的行為可能扮演重要的角色。
URI: http://hdl.handle.net/11455/30357
顯示於類別:昆蟲學系

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