Localization in organelles and function analysis of aquaporin 1 (AQP1) protein in<em> Ectropis obliqua</em> (Lepidoptera: Geometridae)
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ACTA ENTOMOLOGICA SINICA  2017, Vol. 60 Issue (7): 750-759    DOI: 10.16380/j.kcxb.2017.07.003
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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)
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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.
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LI Liang-De
WANG Ding-Feng
LIU Feng-Jing
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.
http://www.insect.org.cn/EN/10.16380/j.kcxb.2017.07.003      or     http://www.insect.org.cn/EN/Y2017/V60/I7/750
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