马氏珠母贝PfAQP4免疫功能初探

doi:10.11978/2020050

Abstract

Abstract:

To identify the immunity function of Aquaporin of Pinctada fucata martensii (PfAQP4), the mRNA expression patterns of PfAQP4 and immune-related genes were detected by qPCR after immunostimulation and RNA interference. The results showed that the mRNA expression of PfAQP4 in the mantle was significantly up-regulated at 24 h (p < 0.05), while that in the digestive gland was significantly down-regulated at 12 h, and increased at 24 h, 36 h and 48 h after LPS injection (p < 0.05). After poly (I : C) injection, the expression of PfAQP4 was significantly up-regulated in the mantle at 36 h and 48 h (p < 0.05), and up-regulated at 48 h in the digestive gland (p < 0.05). When PfAQP4 was knocked down by RNA interference, the mRNA expression of CuZn-SOD in the mantle was significantly down-regulated (p < 0.05); a significant positive correlation of expression level between CuZn-SOD and PfAQP4 was found (r = 0.818, p < 0.001). We showed that PfAQP4 plays an important role in the immune response of P. f. martensii, which provide new data for the innate immunity of P. f. martensii.

Key words: Pinctada fucata martensii, aquaporin 4, immunoregulation, RNA interference, gene expression

CLC Number:

P735.4

PAN Xiaolan, LIU Huiru, XU Meng, XU Hanzhi, ZHANG Hua, HE Maoxian. Preliminary study on immunity function of aquaporin AQP4 in the pearl oyster Pinctada fucata martensii[J].Journal of Tropical Oceanography, 2021, 40(2): 83-89.

Figures/Tables 4

Tab. 1

Fig. 1

Fig. 2

Tab. 2

References 33

[1]	高沿, 2016. 凡纳滨对虾水孔蛋白-4基因克隆、表达及其对盐度胁迫的反应研究[D]. 北京: 中国科学院大学(in Chinese).
[2]	潘肖兰, 刘惠茹, 许濛, 等, 2020. 马氏珠母贝水通道蛋白基因AQP4 cDNA克隆和表达分析[J]. 热带海洋学报, 39(3):66-75.
	PAN XIAOLAN, LIU HUIRU, XU MENG, et al, 2020. Cloning and expression analysis of aquaporin gene AQP4 cDNA from Pinctada fucata martensii[J]. Journal of Tropical Oceanography, 39(3):66-75 (in Chinese with English abstract).
[3]	孙乐, 倪云翔, 丁之德, 2016. 宫内环境变化对子代大脑海马发育影响的分子机制[J]. 国际妇产科学杂志, 43(5):547-551, 560.
	SUN LE, NI YUNXIANG, DING ZHIDE, 2016. The epigenetic influence of intrauterine environment changes on offspring′s hippocampal developmen[J]. Journal of International Obstetrics and Gynecology, 43(5):547-551, 560 (in Chinese with English abstract).
[4]	王晶, 李红玲, 赵龙, 等, 2018. 不同疗程高压氧治疗对脑出血大鼠血肿周围AQP4和SOD表达的影响[J]. 中国康复医学杂志, 33(1):29-35.
	WNAG JING, LI HONGLING, ZHAO LONG, et al, 2018. Effects of different hyperbaric oxygen treatment courses on perihematomal edema and expression of aquaporin-4 and superoxide dismutase in rats with intracerebral hemorrhage[J]. Chinese Journal of Rehabilitation Medicine, 33(1):29-35 (in Chinese with English abstract).
[5]	武柠子, 马慧萍, 王宁, 等, 2016. 脑水肿分子机制的研究进展[J]. 解放军医药杂志, 28(6):14-18.
	WU NINGZI, MA HUIPING, WANG NING, et al, 2016. Progression on molecular mechanisms of cerebral edema[J]. Medical & Pharmaceutical Journal of Chinese People’s Liberation Army, 28(6):14-18 (in Chinese with English abstract).
[6]	袁牧, 王昌留, 王一斐, 等, 2016. 超氧化物歧化酶的研究进展[J]. 中国组织化学与细胞化学杂志, 25(6):550-558.
	YUAN MU, WANG CHANGLIU, WANG YIFEI, et al, 2016. Progress in the research of superoxide dismutase[J]. Chinese Journal of Histochemistry and Cytochemistry, 25(6):550-558 (in Chinese with English abstract).
[7]	朱丹, 李宏俊, 高祥刚, 等, 2010. 文蛤胞内铜锌超氧化物歧化酶基因的克隆与序列分析[J]. 生物技术通报,(11): 123-128, 133.
	ZHU DAN, LI HONGJUN, GAO XIANGGANG, et al, 2010. Molecular cloning and sequence analysis of an intracellular Cu/Zn-superoxide dismutase gene from Hard Clam (Meretrix meretrix)[J]. Biotechnology Bulletin, (11): 123-128, 133 (in Chinese with English abstract).
[8]	ADZIGBLI L, YU WANMEI, LI JUNHUI, et al, 2019. Influence of age on pearl production performance, enzymatic activity, and immune-related gene expression of the pearl oyster Pinctada fucata martensii[J]. North American Journal of Aquaculture, 81(4):430-437.
[9]	ADZIGBLI L, WANG ZIMAN, LI JUNHUI, et al, 2020. Survival, retention rate and immunity of the black shell colored stocks of pearl oyster Pinctada fucata martensii after grafting operation[J]. Fish & Shellfish Immunology, 98: 691-698. pmid: 31693946
[10]	BAO YONGBO, LI LI, XU FEI, et al, 2009. Intracellular copper/zinc superoxide dismutase from bay scallop Argopecten irradians: its gene structure, mRNA expression and recombinant protein[J]. Fish & Shellfish Immunology, 27(2):210-220. pmid: 19426808
[11]	BELAYEV L, LU YOUMING, BAZAN N G. 2012. Chapter 35-brain ischemia and reperfusion: cellular and molecular mechanisms in stroke injury[M]//BRADY S T, SIEGEL G J, ALBERS R W, et al, Basic neurochemistry: principles of molecular, cellular, and medical neurobiology. New York: Academic Press: 621-642.
[12]	CAO SHENGLONG, ZHU PING, YU XIAOBO, et al, 2016. Hydrogen sulfide attenuates brain edema in early brain injury after subarachnoid hemorrhage in rats: Possible involvement of MMP-9 induced blood-brain barrier disruption and AQP4 expression[J]. Neuroscience Letters, 621: 88-97. doi: 10.1016/j.neulet.2016.04.018 pmid: 27080433
[13]	DAI WANGSHU, YAN JUNJUN, CHEN GUANGZONG, et al, 2018. AQP4-knockout alleviates the lipopolysaccharide- induced inflammatory response in astrocytes via SPHK1/ MAPK/AKT signaling[J]. International Journal of Molecular Medicine, 42(3):1716-1722. doi: 10.3892/ijmm.2018.3749 pmid: 29956748
[14]	HO J D, YEH R, SANDSTROM A, et al, 2009. Crystal structure of human aquaporin 4 at 1.8 Å and its mechanism of conductance[J]. Proceedings of the National Academy of Sciences of the United States of America, 106(18):7437-7442. pmid: 19383790
[15]	HUA WEI, CHEN XUEMEI, WANG JUNMIN, et al, 2020. Mechanisms and potential therapeutic targets for spontaneous intracerebral hemorrhage[J]. Brain Hemorrhages, 1(2):99-104.
[16]	KIM K Y, LEE S Y, CHO Y S, et al, 2007. Molecular characterization and mRNA expression during metal exposure and thermal stress of copper/zinc-and manganese-superoxide dismutases in disk abalone, Haliotis discus discus[J]. Fish & Shellfish Immunology, 23(5):1043-1059. pmid: 17574439
[17]	LI CHENGHUA, SUN HUILI, CHEN AIQIN, et al, 2010. Identification and characterization of an intracellular Cu, Zn-superoxide dismutase (icCu/Zn-SOD) gene from clam Venerupis philippinarum[J]. Fish & Shellfish Immunology, 28(3):499-503. pmid: 19948222
[18]	LIN Y C, VASEEHARAN B, CHEN J C, 2008. Identification of the extracellular copper-zinc superoxide dismutase (ecCuZnSOD) gene of the mud crab Scylla serrata and its expression following β-glucan and peptidoglycan injections[J]. Molecular Immunology, 45(5):1346-1355. doi: 10.1016/j.molimm.2007.09.005 pmid: 17949815
[19]	LIU YANJUN, TANG XIANGQI, 2018. Depressive syndromes in autoimmune disorders of the nervous system: prevalence, etiology, and influence[J]. Frontiers in Psychiatry, 9: 451. doi: 10.3389/fpsyt.2018.00451 pmid: 30319458
[20]	NELSON P A, KHODADOUST M, PRODHOMME T, et al, 2010. Immunodominant T cell determinants of aquaporin-4, the autoantigen associated with neuromyelitis optica[J]. PLoS One, 5(11):e15050. doi: 10.1371/journal.pone.0015050 pmid: 21151500
[21]	NI DUOJIAO, SONG LINSHENG, GAO QIANG, et al, 2007. The cDNA cloning and mRNA expression of cytoplasmic Cu, Zn superoxide dismutase (SOD) gene in scallop Chlamys farreri[J]. Fish & Shellfish Immunology, 23(5):1032-1042. doi: 10.1016/j.fsi.2007.04.008 pmid: 17574438
[22]	PANNICKE T, WURM A, IANDIEV I, et al, 2010. Deletion of aquaporin-4 renders retinal glial cells more susceptible to osmotic stress[J]. Journal of Neuroscience Research, 88(13):2877-2888. pmid: 20544823
[23]	PAPADOPOULOS M C, VERKMAN A S, 2007. Aquaporin-4 and brain edema[J]. Pediatric Nephrology, 22(6):778-784. pmid: 17347837
[24]	PÉREZ-HERNÁNDEZ M, FERNÁNDEZ-VALLE M E, RUBIO-ARAIZ A, et al, 2017. 3,4-Methylenedioxymeth- amphetamine (MDMA, ecstasy) produces edema due to BBB disruption induced by MMP-9 activation in rat hippocampus[J]. Neuropharmacology, 118: 157-166. doi: 10.1016/j.neuropharm.2017.03.019 pmid: 28322979
[25]	SUN H, LIANG R, YANG B, et al, 2016. Aquaporin-4 mediates communication between astrocyte and microglia: implications of neuroinflammation in experimental Parkinson’s disease[J]. Neuroscience, 317: 65-75. pmid: 26774050
[26]	SUN LIN, LI MAN, MA XUN, et al, 2019. Inhibiting high mobility group box-1 reduces early spinal cord edema and attenuates astrocyte activation and aquaporin-4 expression after spinal cord injury in rats[J]. Journal of Neurotrauma, 36(3):421-435. doi: 10.1089/neu.2018.5642 pmid: 29929431
[27]	ULUSOY C, TÜZÜN E, KÜRTÜNCÜ M, et al, 2012. Comparison of the cytokine profiles of patients with neuronal- antibody-associated central nervous system disorders[J]. International Journal of Neuroscience, 122(6):284-289.
[28]	WANG HONGHAO, ZHONG XIAONAN, WANG KAI, et al, 2012. Interleukin 17 gene polymorphism is associated with anti-aquaporin 4 antibody-positive neuromyelitis optica in the Southern Han Chinese-A case control study[J]. Journal of the Neurological Sciences, 314(1-2):26-28. pmid: 22118860
[29]	WANG LIUHUA, TANG HUA, WANG CHAO, et al, 2019. Aquaporin 4 deficiency alleviates experimental colitis in mice[J]. The FASEB Journal, 33(8):8935-8944. pmid: 31034776
[30]	YANG JINGYU, XUE XUE, TIAN HUA, et al, 2014. Role of microglia in ethanol-induced neurodegenerative disease: pathological and behavioral dysfunction at different developmental stages[J]. Pharmacology & Therapeutics, 144(3):321-337. doi: 10.1016/j.pharmthera.2014.07.002 pmid: 25017304
[31]	ZHANG HUA, OU ZEKUI, XU MENG, et al, 2018. Molecular cloning and characterization of a putative mitogen-activated protein kinase (Erk1/2) gene: Involvement in mantle immunity of Pinctada fucata[J]. Fish & Shellfish Immunology, 80: 63-70. doi: 10.1016/j.fsi.2018.05.047 pmid: 29859309
[32]	ZHAO XIAOXIA, WANG QINGHENG, JIAO YU, et al, 2012. Identification of genes potentially related to biomineralization and immunity by transcriptome analysis of pearl sac in Pearl Oyster Pinctada martensii[J]. Marine Biotechnology, 14(6):730-739. doi: 10.1007/s10126-012-9438-3 pmid: 22351046
[33]	ZHOU YU, LU ZUNENG, GUO YUANJIN, et al, 2010. Favorable effects of MMP-9 knockdown in murine herpes simplex encephalitis using small interfering RNA[J]. Neurological Research, 32(8):801-809. pmid: 20483026

引物名称	引物序列(5' — 3')	用途
dsAQP4-T7F	GGATCCTAATACGACTCACTATAGGTTGCCTTTGGGCTATCGG	PfAQP4 RNAi含T7启动子序列正向引物
dsAQP4-T7R	GGATCCTAATACGACTCACTATAGGCGGAAGGAGCAGCAAAGAC	PfAQP4 RNAi含T7启动子序列反向引物
dsGFP4-T7F	GGATCCTAATACGACTCACTATAGGCGACGTAAACGGCCACAAGTT	GFP RNAi含T7启动子序列正向引物
dsGFP-T7R	GGATCCTAATACGACTCACTATAGGATGGGGGTGTTCTGCTGGTAG	GFP RNAi含T7启动子序列反向引物
dsAQP4-F	TTGCCTTTGGGCTATCGG	PfAQP4 RNAi正向引物
dsAQP4-R	CGGAAGGAGCAGCAAAGAC	PfAQP4 RNAi反向引物
dsGFP-F	CGACGTAAACGGCCACAAGTT	GFP RNAi正向引物
dsGFP-R	ATGGGGGTGTTCTGCTGGTAG	GFP RNAi反向引物
Faqp	CTGTGATGGCGTCAACTGATG	PfAQP4 qPCR正向引物
Raqp	CGCAATCGTGAGAAGATGACC	PfAQP4 qPCR反向引物
F18S	CGTTTCAACAAGACGCCAGTAG	18S qPCR正向引物
R18S	ACGAAAAAAAGGTTTGAGAGACG	18S qPCR反向引物
QT5Rel143	GATGGCAGAGGATGATTCTTCTT	NF-κB qPCR正向引物
QT3Rel332	TGATGGACCTTCACACTCATACC	NF-κB qPCR反向引物
MMPF	TCTGGCTCATGCGTTTTTCC	MMP qPCR正向引物
MMPR	AGGGCATGTCCAATCTCATGAG	MMP qPCR反向引物
QT5IL17seq460	CCAGTCCTCGTAATAAATGTGAACC	IL-17 qPCR正向引物
QT3IL17seq636	CGCTTCCGCTGCTAGATTCTT	IL-17 qPCR反向引物
LAMPF	TGCCTGTCACAATAATAACC	LAMP qPCR正向引物
LAMPR	AGACTCAAAGTAAGACCACCT	LAMP qPCR反向引物
CuZn-SODF	GAAACTGCAAACCCTATA	CuZn-SOD qPCR正向引物
CuZn-SODR	AGACATTCGTCAAACTCA	CuZn-SOD qPCR反向引物

基因名称	相关系数	p值
基质金属蛋白分解酶	0.356	0.212
核因子κB	0.655*	0.011
白细胞介素17	0.515	0.060
溶酶体膜相关糖蛋白	0.674**	0.008
铜锌超氧化物歧化酶	0.818***	0.000

Preliminary study on immunity function of aquaporin AQP4 in the pearl oyster Pinctada fucata martensii

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 4

References 33

Related Articles 10

Metrics

Comments

Recommended 0

[1]	YUAN Xiangcheng, LIANG Yuxian, SONG Yan, YU Xiaolei, HUANG Hui, ZHOU Weihua. Effects of ocean acidification on the calcification and gene expression in coral Acropora hyacinthus^* [J]. Journal of Tropical Oceanography, 2024, 43(3): 40-48.
[2]	LI Ze, WANG Lijuan, ZOU Congcong, SHU Chang, WU Zhihao, ZOU Yuxia, YOU Feng. Expression characteristics and function of Anti-Müllerian hormone receptor Ⅱ gene (amhr2) in Paralichthys olivaceus [J]. Journal of Tropical Oceanography, 2024, 43(1): 94-106.
[3]	XU Hanzhi, ZHANG Hua, XIONG Panpan, HE Maoxian. Differential responses of allometric individuals to immune stimuli in Pinctada fucata martensii [J]. Journal of Tropical Oceanography, 2022, 41(5): 180-188.
[4]	Xiaolan PAN, Huiru LIU, Meng XU, Hanzhi XU, Hua ZHANG, Maoxian HE. Cloning and expression analysis of aquaporin gene AQP4 cDNA from Pinctada fucata martensii [J]. Journal of Tropical Oceanography, 2020, 39(3): 66-75.
[5]	Jiahao NI, Xiaojing ZHU, Yiping JI, Bin ZHOU, Yajun WANG, Shanliang XU, Danli WANG. Effects of breeding density on the growth, metabolic enzyme activity and related gene expression level of juvenile Pampus argenteus [J]. Journal of Tropical Oceanography, 2020, 39(2): 54-64.
[6]	HUANG Jing, PAN Xiaolan, XU Meng, LIU Wenguang, ZHANG Hua, HE Maoxian. Morphological and SNP markers for analysis of genetic structure of hybrid progeny and their parental populations of Pinctada fucata martensii [J]. Journal of Tropical Oceanography, 2019, 38(6): 80-89.
[7]	Fuxuan WANG, Shu XIAO, Zhiming XIANG, Ziniu YU. Molecular cloning and expression analysis of Commd1 under salinity stress in Crassostrea hongkongensis [J]. Journal of Tropical Oceanography, 2017, 36(1): 48-55.
[8]	SHEN Cheng, LIU Chuwu, LIU Li. Heat-induced stress genes Hsp70, Hsp90 and chloroplast psaA and psbA gene expressions of Galaxea astreata endosymbiotic zooxanthella revealing the ability of tolerance and recovery [J]. Journal of Tropical Oceanography, 2016, 35(3): 72-78.
[9]	HUANG Hui, XU Chang-you, YUAN Tao. Research progress on functional genes involved in coral bleaching [J]. Journal of Tropical Oceanography, 2013, 32(4): 43-50.
[10]	ZHANG Li-juan,HE Mao-xian. Effects of growth hormone and insulin-like growth factor- Ⅰ on shell growth related gene expression in Pinctada fucata Gould [J]. Journal of Tropical Oceanography, 2012, 31(2): 96-101.