Localization in organelles and function analysis of aquaporin 1 (AQP1) protein in<em> Ectropis obliqua</em> (Lepidoptera: Geometridae)
昆虫学报
       Home | About Journal | Editorial Board | Subscription | Instruction | Advertisement | Contact Us | Chinese
ACTA ENTOMOLOGICA SINICA  2017, Vol. 60 Issue (7): 750-759    DOI: 10.16380/j.kcxb.2017.07.003
RESEARCH PAPERS Current Issue | Next Issue | Archive | Adv Search Previous Articles  |  Next Articles  
Localization in organelles and function analysis of aquaporin 1 (AQP1) protein in Ectropis obliqua (Lepidoptera: Geometridae)
LI Liang-De#, WANG Ding-Feng#, LIU Feng-Jing, ZHANG Hui, LI Hui-Ling, WU Guang-Yuan*
(Tea Research Institute, Fujian Academy of Agricultural Sciences, Fu′an, Fujian 355015, China)
 Download: PDF (3859 KB)   HTML (1 KB)   Export: BibTeX | EndNote (RIS)      Supporting Info
Abstract 【Aim】 Aquaporin (AQP) is one of the transmembrane proteins widely existing in cell biological membrane system, and plays an important role in the balance of water osmotic pressure. This study aims to understand the localization of AQP1 in organelles based on the previously cloned full-length cDNA in Ectropis obliqua Prout (EoAQP1), and to explore its effects on cell morphology and cell proliferation. 【Methods】 The eukaryotic expression vectors of green fluorescent protein (GFP) and its fusion protein with EoAQP1 (EoAQP1-GFP) were constructed by double enzyme digestion. Cell expression characteristics of GFP and EoAQP1-GFP proteins in Drosophila melanogaster embryonic cell line S2 were observed by fluorescence microscope and confocal scanning laser microscope. The localization of EoAQP1 protein in organelles in S2 cells was detected by Lyso-Tracker Red and Golgi-Tracker Red kit. The effects of EoAQP1 on cell granularity, size and proliferation were detected by flow cytometry and microplate reader. 【Results】 The eukaryotic expression vectors of GFP and EoAQP1GFP were constructed successfully, and named pAc5.1-GFP and pAc5.1-EoAQP1-GFP, respectively. Fluorescence microscope and confocal scanning laser microscope observation showed that GFP protein was evenly filled in S2 cells and showed stable expression. The EoAQP1-GFP protein was expressed by two types: one showed scattered expression in the cytoplasm and around the nucleus, in which the EoAQP1-GFP protein was in spherical and spheroidic structure similar to lysosome; the other showed the hemisphere and arched expression in the cytoplasm, in which the EoAQP1-GFP protein was in hemisphere and arch structure similar to the Golgi apparatus. Lyso-Tracker Red and Golgi-Tracker Red kit detection showed that EoAQP1 protein had no expression in lysosome but completely overlapped in the Golgi apparatus. Flow cytometry and microplate reader detection indicated that the overexpression of EoAQP1 protein in S2 cells enhanced cell volume and granularity significantly, but had no significant effect on cell proliferation. 【Conclusion】 The EoAQP1 protein is localized and plays functions in the Golgi apparatus. It can change cell morphology, but not promote cell growth.
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
LI Liang-De
WANG Ding-Feng
LIU Feng-Jing
ZHANG Hui
LI Hui-Ling
WU Guang-Yuan
Key wordsEctropis obliqua; aquaporin   lysosome   Golgi apparatus   cell morphology   cell proliferation     
Cite this article:   
LI Liang-De,WANG Ding-Feng,LIU Feng-Jing et al. Localization in organelles and function analysis of aquaporin 1 (AQP1) protein in Ectropis obliqua (Lepidoptera: Geometridae)[J]. ACTA ENTOMOLOGICA SINICA, 2017, 60(7): 750-759.
URL:  
http://www.insect.org.cn/EN/10.16380/j.kcxb.2017.07.003      or     http://www.insect.org.cn/EN/Y2017/V60/I7/750
 
No references of article
[1] XIE Gui-Ying, CHEN Jun-Hua, TANG Qing-Bo, YIN Jian, ZHAO Xin-Cheng. Anatomical structure of the brain of larval Ectropis obliqua (Lepidoptera: Geometridae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(8): 831-838.
[2] LI Liang-De, WANG Ding-Feng, LIU Feng-Jing, LI Hui-Ling, ZHANG Hui, WU Guang-Yuan. cDNA cloning, preparation of polyclonal antibody and subcellular localization of aquaporin 1 (AQP1) in Ectropis obliqua (Lepidoptera: Geoqmetridae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(4): 382-391.
[3] LI Liang-De, WANG Ding-Feng, LI Hui-Ling, ZHANG Hui, WU Guang-Yuan. Molecular cloning, bioinformatics analysis and expression profiling of ecdysteroid receptor gene Eo-EcR in Ectropis obliqua (Lepidoptera: Geometridae)[J]. ACTA ENTOMOLOGICA SINICA, 2015, 58(10): 1063-1071.
[4] XI Yu, YIN Kun-Shan, TANG Mei-Jun, XIAO Qiang. Geographic populations of the tea geometrid, Ectropis obliqua (Lepidoptera: Geometridae) in Zhejiang, eastern China have differentiated into different species[J]. ACTA ENTOMOLOGICA SINICA, 2014, 57(9): 1117-1122.
[5] LIU Hai-Yuan, SHU Ben-Shui, JIANG Chun-Lai, LI Liang-De, ZHONG Guo-Hua. Molecular cloning, characterization and expression analysis of aquaporin 1 (AQP1) gene in Spodoptera litura (Lepidoptera: Noctuidae)[J]. ACTA ENTOMOLOGICA SINICA, 2013, 56(4): 339-349.
[6] MENG Xiang, Hu Jun-Jie, Jin Feng-Liang, Ren Shun-Xiang. Inhibition of cellular proliferation and apoptosis in Spodoptera litura (Lepidoptera: Noctuidae) SL-1 cells induced by destruxin A and destruxin B[J]. ACTA ENTOMOLOGICA SINICA, 2011, 54(9): 1003-1009.
[7] Anirban BASU MAJUMDER, Sunil Kumar PATHAK, Tapan Kumar HATH. Growth and survival of a geometrid defoliator Hyposidra infixaria Walker (Lepidoptera: Geometridae) on four clonal varieties of tea[J]. ACTA ENTOMOLOGICA SINICA, 2011, 54(7): 815-819.
[8] LIN Mei-Juan, XIE Jian, ZHANG Jia-Min, YE Shan, HU Yuan-Yang. Construction of the full-length cDNA clone of Ectropis oblique picorna-like virus[J]. ACTA ENTOMOLOGICA SINICA, 2010, 53(7): 818-823.
[9] . EAG and behavioural responses of Apanteles sp. (Hymenoptera: Braconidae) parasitizing tea geometrids by volatiles from tea shoots[J]. ACTA ENTOMOLOGICA SINICA, 2009, 52(11): 1191-1198.
[10] LI Wen-Ou. Mechanism of cell toxicity of azadirachtin A to Trichoplusia ni Hübner Hi-5 cells[J]. ACTA ENTOMOLOGICA SINICA, 2008, 51(8): 824-829.
[11] WANG Xiao-Chun1,2, ZHANG Jia-Min1, JIANG Hong1, YU Hai-Yang1, TAN-Li1, HU Yuan-Yang1*. Cloning and sequencing of the 5'untranslation region of Ectropis oblique picornavirus with a comparison to that of mammalian picornaviruses[J]. ACTA ENTOMOLOGICA SINICA, 2004, 47(5): 573-578.
[12] Xu Ning,Chen Zong-mao,You Xiao-qing. SOLATION AND IDENTIFICATION OF TEA PLANT VOLATILES ATTRACTIVE TO TEA GEOMETRID PARASITOIDS[J]. ACTA ENTOMOLOGICA SINICA, 1999, 42(2): 126-131.
Copyright © 2011 ACTA ENTOMOLOGICA SINICA
Supported by:Beijing Magtech