Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression

doi:10.3969/j.issn.1004-1524.20230104

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (2): 247-253.DOI: 10.3969/j.issn.1004-1524.20230104

• Animal Science • Previous Articles Next Articles

Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression

PENG Jiacheng¹(), WU Yue¹, XU Jiehao², XIA Meiwen¹, QI Tianpeng¹, XU Haisheng¹^,³^,^*()

1. College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
2. College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, Zhejiang, China
3. Zhejiang University Huzhou South Taihu Lake Modern Agricultural Science and Technology Promotion Center, Huzhou 313000, Zhejiang, China

Received:2023-02-06 Online:2024-02-25 Published:2024-03-05

Abstract

Abstract:

Cadmium (Cd), with strong accumulation and toxicity, is one of the non-essential heavy metal elements, which could cause great harm to aquatic organisms. To better understand the molecular toxicity of cadmium to Macrobrachium nipponense, and provide biomarkers for ecological risk warning and assessment of cadmium pollution in water environment. In this paper, paxillin gene (MnPXN) of M. nipponense was cloned, and the effect of cadmium stress on its expression was examined. The total length of MnPXN gene is 3 655 bp, including 1 491 bp coding region, encoding 496 amino acids; 6 low complexity regions and 3 conservative LIM domains are included in MnPXN, forming a branch with Litopenaeus vannamei. It is closely related to insects such as silkworm and Tribolium castaneum, and is farthest related to nematodes. The results of quantitative reverse transcriptase-mediated PCR (qRT-PCR) showed that the expression level of MnPXN was high in hepatopancreas, stomach and gill, but low in heart and muscle. The expression level of MnPXN gene in hepatopancreas that after 24 h of treatment with 0.242 7 mg·L^-1 CdCl₂ was significantly higher than that in control group; After 6 h of 0.020 0 mg·L^-1 CdCl₂ stress, the expression level of MnPXN gene increased significantly, reaching the maximum at 24 h, and gradually recovered to the initial level at 96 h. Besides, RNA interference could significantly reduce the expression level of MnPXN in hepatopancreas and significantly increase the mortality of M. nipponense, indicating that MnPXN could reduce the toxic effect of cadmium stress on M. nipponense, and its mechanism needs further study.

Key words: cadmium stress, Macrobrachium nipponense, paxillin, gene expression, RNA interference

CLC Number:

S917.4

PENG Jiacheng, WU Yue, XU Jiehao, XIA Meiwen, QI Tianpeng, XU Haisheng. Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression[J]. Acta Agriculturae Zhejiangensis, 2024, 36(2): 247-253.

Figures/Tables 6

References 20

[1]	LÓPEZ-COLOMÉ A M, LEE-RIVERA I, BENAVIDES-HIDALGO R, et al. Paxillin: a crossroad in pathological cell migration[J]. Journal of Hematology & Oncology, 2017, 10(1): 50.
[2]	YANG W J, ZHONG J, YU J G, et al. The structure and functions of paxillin and its roles in neovascularization[J]. European Review for Medical and Pharmacological Sciences, 2017, 21(8): 1768-1773.
[3]	TURNER C E. Molecules in focus Paxillin[J]. The International Journal of Biochemistry & Cell Biology, 1998, 30(9): 955-959.
[4]	DEAKIN N O, TURNER C E. Paxillin comes of age[J]. Journal of Cell Science, 2008, 121(Pt 15): 2435-2444.
[5]	ZHOU J X, LIU T E, XU H J, et al. LncRNA FIRRE promotes the proliferation and metastasis of hepatocellular carcinoma by regulating the expression of PXN through interacting with MBNL3[J]. Biochemical and Biophysical Research Communications, 2022, 625: 188-195.
[6]	SALMAN S. The invasion of Macrobrachium nipponense(De Haan, 1849) (Caridea: Palaemonidae) into the Southern Iraqi Marshes[J]. Aquatic Invasions, 2006, 1(3): 109-115.
[7]	KRIVOHLAVEK A, KUHARIĆ Ž, MARJANOVIĆ ČERMAK A M, et al. Assessment of intracellular accumulation of cadmium and thallium[J]. Journal of Pharmacological and Toxicological Methods, 2021, 110: 107087.
[8]	NAIKOO M I, RAGHIB F, DAR M I, et al. Uptake, accumulation and elimination of cadmium in a soil-Faba bean (Vicia faba)-Aphid (Aphis fabae)-Ladybird (Coccinella transversalis) food chain[J]. Chemosphere, 2021, 279: 130522.
[9]	LIU Y N, CHEN Q Q, LI Y Q, et al. Toxic effects of cadmium on fish[J]. Toxics, 2022, 10(10): 622.
[10]	ZHANG H, REYNOLDS M. Cadmium exposure in living organisms: a short review[J]. The Science of the Total Environment, 2019, 678: 761-767.
[11]	BOSCH A C, O’NEILL B, SIGGE G O, et al. Heavy metals in marine fish meat and consumer health: a review[J]. Journal of the Science of Food and Agriculture, 2016, 96(1): 32-48.
[12]	KIM M S, YOO N J, LEE S H. Absence of paxillin gene mutation in lung cancer and other common solid cancers[J]. Tumori, 2011, 97(2): 211-213.
[13]	MAZAKI Y, UCHIDA H, HINO O, et al. Paxillin isoforms in mouse: lack of the gamma isoform and developmentally specific beta isoform expression[J]. The Journal of Biological Chemistry, 1998, 273(35): 22435-22441.
[14]	CRAWFORD B D, HENRY C A, CLASON T A, et al. Activity and distribution of paxillin, focal adhesion kinase, and cadherin indicate cooperative roles during zebrafish morphogenesis[J]. Molecular Biology of the Cell, 2003, 14(8): 3065-3081.
[15]	GAO J, HUANG M, LAI J J, et al. Kindlin supports platelet integrin αIIbβ3 activation by interacting with paxillin[J]. Journal of Cell Science, 2017, 130(21): 3764-3775.
[16]	刘长锁, 陈乃宏. 桩蛋白的结构与功能[J]. 生命的化学, 2005, 25(3): 208-210.
	LIU C S, CHEN N H. Structure and function of pilin[J]. Chemistry of Life, 2005, 25(3): 208-210. (in Chinese)
[17]	吴迪, 王梦圆, 史永富, 等. 镉在甲壳类水生生物中的蓄积现状及赋存形态研究进展[J]. 核农学报, 2023, 37(1): 128-139.
	WU D, WANG M Y, SHI Y F, et al. Research progress of cadmium accumulation and speciation in crustacean aquatic organisms[J]. Journal of Nuclear Agricultural Sciences, 2023, 37(1): 128-139. (in Chinese with English abstract)
[18]	SHEN X D, TANG Z Q, BAI Y, et al. Astragalus polysaccharide protects against cadmium-induced autophagy injury through reactive oxygen species (ROS) pathway in chicken embryo fibroblast[J]. Biological Trace Element Research, 2022, 200(1): 318-329.
[19]	WEI Z X, SHAIKH Z A. Cadmium stimulates metastasis-associated phenotype in triple-negative breast cancer cells through integrin and β-catenin signaling[J]. Toxicology and Applied Pharmacology, 2017, 328: 70-80.
[20]	CHOONG G, LIU Y, TEMPLETON D M. Interplay of calcium and cadmium in mediating cadmium toxicity[J]. Chemico-Biological Interactions, 2014, 211: 54-65.

引物 Primer	引物序列 Primer sequence(5'→3')	用途 Purpose
MnPXN-3'-F₁	CAATGCCAGCAGCCTCCAGACAC	RACE
MnPXN-3'-R₁	AAGCCGTCCTCGCCGAAGGTGTT	RACE
MnPXN-3'-F₂	TGGACGAGTTGAACAGCGTTGACA	RACE
MnPXN-3'-R₂	TCCCTTCTGGGTCGTCGTGATGC	RACE
MnPXN-3'-F₃	GAATGACCTGAGTGGCACCTGAC	RACE
MnPXN-3'-R₃	TGCCGTGAAGAATGACATAGAAT	RACE
MnPXN-3'-F₄	GAGGAAGTGCGGACGGAGCAGAGGT	RACE
MnPXN-3'-R₄	GATCTCGATGTGCCGTGAAGAAT	RACE
MnPXN-3'-F₅	GTTATGTCATTGGCTCAAGAGGA	RACE
MnPXN-3'-R₅	GTGTATTTATACTGTATGCCTAACC	RACE
MnPXN-3'-F₆	GCTACAAGAGGCTCCCAACTACAT	RACE
MnPXN-3'-R₆	CAGGAGGTAAAGACTACTGATGT	RACE
MnPXN-3'-F₇	AAAAGGTTTGGTAAATGTCCAGGTT	RACE
MnPXN-3'-R_UPM	CTAATACGACTCACTATAGGGC	RACE
MnPXN-5'-F₁	TTGTTCATCCAGGAGGTAAAGAC	RACE
MnPXN-5'-R₁	GCCAATAAGGCATCAATGGTATCCTG	RACE
MnPXN-5'-F₂	GAAGTTTGGCAGTATCTTATGGA	RACE
MnPXN-5'-R₂	TTGTGAATGAGGGTGGCGTTTGG	RACE
MnPXN-5'-F₃	CAATAAGGCATCAATGGTATCCTGCA	RACE
MnPXN-5'-R₃	AATTTCAAAAATCTAATT	RACE
MnPXN-5'-F_UPM	CTAATACGACTCACTATAGGGCAA GCAGTGGTATCAACGCAGAGT	RACE
MnPXN-5'-R₄	GGGCATCATTCTTTGTCTTTCTT	RACE
MnPXN-RT-F	CCCCATCTTGGATAAATGCGTGAC	qRT-PCR
MnPXN-RT-R	AAGCCGTCCTCGCCGAAGGTGTTGC	qRT-PCR
β-actin-F	GTGCCCATCTACGAGGGTTA	qRT-PCR
β-actin-R	CGTCAGGGAGCTCGTAAGAC	qRT-PCR

引物 Primer	引物序列 Primer sequence(5'→3')	用途 Purpose
MnPXN-3'-F₁	CAATGCCAGCAGCCTCCAGACAC	RACE
MnPXN-3'-R₁	AAGCCGTCCTCGCCGAAGGTGTT	RACE
MnPXN-3'-F₂	TGGACGAGTTGAACAGCGTTGACA	RACE
MnPXN-3'-R₂	TCCCTTCTGGGTCGTCGTGATGC	RACE
MnPXN-3'-F₃	GAATGACCTGAGTGGCACCTGAC	RACE
MnPXN-3'-R₃	TGCCGTGAAGAATGACATAGAAT	RACE
MnPXN-3'-F₄	GAGGAAGTGCGGACGGAGCAGAGGT	RACE
MnPXN-3'-R₄	GATCTCGATGTGCCGTGAAGAAT	RACE
MnPXN-3'-F₅	GTTATGTCATTGGCTCAAGAGGA	RACE
MnPXN-3'-R₅	GTGTATTTATACTGTATGCCTAACC	RACE
MnPXN-3'-F₆	GCTACAAGAGGCTCCCAACTACAT	RACE
MnPXN-3'-R₆	CAGGAGGTAAAGACTACTGATGT	RACE
MnPXN-3'-F₇	AAAAGGTTTGGTAAATGTCCAGGTT	RACE
MnPXN-3'-R_UPM	CTAATACGACTCACTATAGGGC	RACE
MnPXN-5'-F₁	TTGTTCATCCAGGAGGTAAAGAC	RACE
MnPXN-5'-R₁	GCCAATAAGGCATCAATGGTATCCTG	RACE
MnPXN-5'-F₂	GAAGTTTGGCAGTATCTTATGGA	RACE
MnPXN-5'-R₂	TTGTGAATGAGGGTGGCGTTTGG	RACE
MnPXN-5'-F₃	CAATAAGGCATCAATGGTATCCTGCA	RACE
MnPXN-5'-R₃	AATTTCAAAAATCTAATT	RACE
MnPXN-5'-F_UPM	CTAATACGACTCACTATAGGGCAA GCAGTGGTATCAACGCAGAGT	RACE
MnPXN-5'-R₄	GGGCATCATTCTTTGTCTTTCTT	RACE
MnPXN-RT-F	CCCCATCTTGGATAAATGCGTGAC	qRT-PCR
MnPXN-RT-R	AAGCCGTCCTCGCCGAAGGTGTTGC	qRT-PCR
β-actin-F	GTGCCCATCTACGAGGGTTA	qRT-PCR
β-actin-R	CGTCAGGGAGCTCGTAAGAC	qRT-PCR

Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 20

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	TANG Jinyu, QIN Baoli, YE Jianyong, DAI Yangxin. Effects of stocking modes on growth traits and muscle nutritional composition of Macrobrachium nipponense [J]. Acta Agriculturae Zhejiangensis, 2024, 36(2): 254-263.
[2]	LIU Xiaolin, SUN Tingting, YANG Jie, HE Hengbin. Cloning and expression analysis of FLS gene of flavonol synthetase in Lilium auratum and L.speciosum var. gloriosoides [J]. Acta Agriculturae Zhejiangensis, 2024, 36(2): 344-357.
[3]	LIANG Feishuang, LIANG Huafang, Huang Jiayu, WANG Panmei, WEN Chongqing. Effect of RNA interference with PhCatC1/2 gene on expression of related immune genes in Panulirus homarus [J]. Acta Agriculturae Zhejiangensis, 2023, 35(5): 1037-1047.
[4]	WU Jiao, GONG Chengyu, CHEN Chaoqun, CHEN Hongxu, LIU Junhong, TANG Wenjing, CHU Yuanqi, YANG Wenlong, ZHANG Yao, GONG Ronggao. Ecological response of organic acid metabolism of Huangguogan fruits at different altitudes [J]. Acta Agriculturae Zhejiangensis, 2023, 35(4): 853-861.
[5]	LI Hongyi, ZHOU Runsheng, LIANG Xiaoling, ZHANG Chuyue, LYU Qixin, YANG Changhua, ZHANG Mao. Effect of dietary calcium and phosphorus level on Magang geese growth performance and liver gene expression [J]. Acta Agriculturae Zhejiangensis, 2023, 35(11): 2533-2542.
[6]	KONG Fanwang, ZHANG Zhigang, LI Wei, CHEN Yufeng, WANG Changjiang, ZHENG Yaqin, XU Meng. Identification of 4CL gene family in peach and its expression analysis in fruit coloration during development stage and chilling injury during post-harvest low temperature storage [J]. Acta Agriculturae Zhejiangensis, 2023, 35(11): 2600-2610.
[7]	LIN Xianyu, LI Ziqian, BAI Song, LUO Jun, QU Yan. Changes of antioxidant enzyme activity and differential expression of key genes in Camellia reticulata during drought-rehydration process [J]. Acta Agriculturae Zhejiangensis, 2023, 35(11): 2611-2620.
[8]	FAN Liying, FAN Tingting, TONG Zongjun, LIANG Liyun, ZHAO Zhiyong, CHEN Hui, ZHOU Changyan, ZHAO Xiaoyan. Effects on accumulation of cadmium and antioxidant system of different Morchella spp. under cadmium stress [J]. Acta Agriculturae Zhejiangensis, 2023, 35(10): 2321-2331.
[9]	XIONG Xingwei, WANG Yiqin, TIAN Huaizhi, ZHANG Suqin, GENG Guangdong. Molecular mechanisms of chlorophyll-reduced cotyledon based on transcriptome sequencing in pumpkin [J]. Acta Agriculturae Zhejiangensis, 2023, 35(1): 90-102.
[10]	YAN Mei, YAO Yandong, MOU Kaiping, DAN Yuanyuan, LI Weitai, LIAO Weibiao. Involvement of abscisic acid in hydrogen gas-enhanced drought resistance by improving antioxidant enzyme activity and gene expression in tomato seedlings [J]. Acta Agriculturae Zhejiangensis, 2022, 34(9): 1901-1910.
[11]	JIANG Haoliang, HUANG Yun, LIANG Shaofang, XIE Mengchen, XU Tiancheng, SONG Zhiting, XIANG Wenwen, CHEN Qingchun, WAN Xiaorong, SUN Wei. Influences of cadmium stress on seedling growth of different sweet corn inbred lines and screening of associated molecular markers via simple sequence repeats [J]. Acta Agriculturae Zhejiangensis, 2022, 34(8): 1582-1590.
[12]	DONG Yuanyuan, XU Heng, ZHANG Hua, ZHANG Heng, WANG Fulin, GU Nana, ZHU Ying. Dynamic profile of genes related to seed dormancy under high humidity condition during late stage of rice grain filling [J]. Acta Agriculturae Zhejiangensis, 2022, 34(6): 1103-1113.
[13]	DU Hong, LI Yupeng, CHENG Wen, XIAO Rongying, HU Peng. Effects of arbuscular mycorrhizal fungi on plant roots and soil microenvironment under cadmium stress [J]. Acta Agriculturae Zhejiangensis, 2022, 34(5): 1039-1048.
[14]	YU Yanling, LUO Honglin, LUO Hui, FENG Pengfei, PAN Chuanyan, SONG Manling, XIAO Rui, ZHANG Yongde. Genome-wide identification and expression of MRF gene family in embryonic development of Trachinotus ovatus [J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 695-705.
[15]	LIU Tongjin, XU Mingjie, WANG Jinglei, LIU Liangfeng, CUI Qunxiang, BAO Chonglai, WANG Changyi. Genome-wide identification and expression analysis of ALMT gene family in radish [J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 746-755.