Elsevier

Fish & Shellfish Immunology

Volume 106, November 2020, Pages 975-981
Fish & Shellfish Immunology

Full length article
New insights into the immune regulation and tissue repair of Litopenaeus vannamei during temperature fluctuation using TMT-based proteomics

https://doi.org/10.1016/j.fsi.2020.09.014Get rights and content

Highlights

  • The first TMT-based proteomic profiling for shrimp during temperature fluctuation.

  • Shrimp could cope with cold stress by metabolic inhibition.

  • Shrimp up-regulated intestinal immune-related proteins to maintain homeostasis during temperature fluctuation.

Abstract

To investigate shrimp immunoregulation and tissue self-repair mechanism during temperature fluctuation stage, Litopenaeus vannamei (L. vannamei) was treated under conditions of gradual cooling from an acclimation temperature (28 °C, C group) to 13 °C (T group) in 2 days with a cooling rate of 7.5 °C/d and then rewarmed to 28 °C (R group) with the same rate. Tandem mass tags (TMT) -based proteomics technology was used to investigate the protein abundance changes of intestine in L. vannamei during temperature fluctuation. The results showed that a total of 5796 proteins with function annotation were identified. Of which, the abundances of 1978 proteins (34%) decreased after cooling and then increased after rewarming, 1498 proteins (26%) increased during the whole stage, 1263 proteins (22%) increased after cooling and then decreased after rewarming and 1057 proteins (18%) decreased during the whole stage. Differentially expressed proteins such as C-lectin, NFκBIA and Caspase may contributed to the regulation of immunity and tissue repair of shrimp intestine during the temperature fluctuation stage. These findings contribute to the better understanding of shrimp’ regulatory mechanism against adverse environment.

Introduction

Temperature is one of the considerable environmental stimuli in aquaculture. So far, some studies focused their interests on the influence of short-term extreme weather events, demonstrating that it has directly affected the physiological performance, growth, and survival of plant [1], animal [2] and human beings [3,4]. However, the impact of short extreme weather on aquatic animals still needs further study. Litopenaeus vannamei (L. vannamei) is one of the most essential global economic aquaculture shrimps. However, environmental factors such as changes in oxygen content [5], pH [6], salinity [7] and pollutants like sulfide [8] and ammonia [9] can directly affect its health. Besides, since it comes from the tropics, L. vannamei is vulnerable to cold stress [10,11]. Guangdong is one of the main areas of shrimp culture in China. But the annual cold wave of winter in Guangzhou has threatened the shrimp culture industry in China for a long time and caused huge economic losses. It has been reported that L. vannamei might appear the phenomena such as growth arrest, stop swimming and feeding with the water temperature decreased, and L. vannamei could even death after temperature fell below 13 °C [12,13].

Meanwhile, our previous study found that L. vannamei intestine has a self-repairing ability and it may mobilize the immune defense system to maintain homeostasis during temperature fluctuation with a changing rate of 7.5 °C/d [14,15]. However, the adaptation mechanism of the shrimp during temperature fluctuation still requires further study. Understanding how shrimp regulates the immunity and self-repairing ability may help us protecting shrimp from cold stress.

Proteins are considered as the ultimate executors to carry out biological functions. With the rapid development of the next generation sequencing in recent years, proteomics analysis is widely used to help us understanding the response of aquatic animals to various environmental stimuli [16,17]. For shrimp, the protein response of shrimp to some environmental stimuli such as ammonia [18] and salinity [19] stress has been studied. While more proteomics studies mainly focused on the response to some directly pathogens stimuli, such as white spot syndrome virus infection to kuruma shrimp [20] and Chinese shrimp [21], microsporidian infection to L. vannamei [22]. Isobaric tags for relative and absolute quantification (iTRAQ) and tandem mass tags (TMT) labeling plus liquid chromatography–mass spectrometry (LC-MS/MS) are two typical approach for protein detection and quantification. But TMT-based proteomics analysis can quantify more samples at the same time than iTRAQ. So far, the homeostasis mechanism of L. vannamei using TMT during temperature fluctuation stage has not been reported.

In the present study, TMT-based quantitative proteomics was used to investigate the protein response during temperature fluctuation stage. The findings could provide valuable reference for finding feasible ways to protect intestine health against adverse environment.

Section snippets

Experimental shrimp and culture conditions

The experimental shrimp were collected from a commercial farm in Panyu (Guangdong, China). Before experiments, the shrimp were acclimated in diluted seawater tanks with air-pumped circulating at least 7 days in lab. The salinity of the rearing water was 5‰ and the temperature was 28 ± 1 °C. Commercial shrimp feeds (Haida Feed, China) with a ratio of 5% of shrimp body weight was given two times per day. This condition was consistent with that of the commercial farm.

Treatment and sampling

A total of sixty-six healthy

Statistic of protein profiling

A total of 533,945 spectrums were obtained from LC-MS/MS data. After database searching by Proteome Discoverer Software, 5880 proteins were identified based on Pacific white shrimp genome reference (Fig. 1A). Among all the identified proteins, the protein molecular weight of 85.12% proteins was ranged from 1–101 kDa (Figs. 1B) and 75.15% of the protein's sequences coverage more than 5% (Fig. 1C), which indicated a good sequence coverage of these identified proteins.

Cluster and functional annotation analysis

During temperature

Response to cope with temperature fluctuation

Temperature is a major environmental factor that affects growth and survival of aquatic animal. Understanding how shrimp cope with the stress of temperature fluctuation is important for shrimp breeding industry. Previous study has found that red claw crayfish (Cherax quadricarinatus) can down-regulated metabolism under low temperature [37]. In this study, the proportion of “energy metabolism” ranks first and “oxidative phosphorylation” was the most significant pathway in T13L group. Animals

Conclusions

This study provides the first quantitative analysis in L. vannamei protein changes during temperature fluctuation stage by TMT proteomics. We deduce that shrimp down-regulated energy metabolism to cope with cold-stress, while it might resist cold though digestion and absorption of protein. In addition, shrimp might up-regulated intestinal immune-related proteins to maintain homeostasis during temperature fluctuation. DEPs such as NFκBIA and Caspase contributed to the regulation of immunity and

CRediT authorship contribution statement

Zhenlu Wang: Conceptualization, Investigation, Writing - original draft. Lei Wang: Software, Data curation. Jiang Zhou: Validation. Jixing Zou: Resources, Supervision. Lanfen Fan: Supervision.

Acknowledgments

This work was supported by the National Natural Science Foundation of China (31600322).

References (65)

  • X. Dong et al.

    Proteomic profile and toxicity pathway analysis in zebrafish embryos exposed to bisphenol A and di-n-butyl phthalate at environmentally relevant levels

    Chemosphere

    (2018)
  • X. Lu et al.

    iTRAQ-based comparative proteome analysis for molecular mechanism of defense against acute ammonia toxicity in Pacific White shrimp Litopenaeus vannamei

    Fish Shellfish Immunol.

    (2018)
  • C. Xu et al.

    Comparative proteome analysis of the hepatopancreas from the Pacific white shrimp Litopenaeus vannamei under long-term low salinity stress

    J. Proteomics

    (2017)
  • X. Ren et al.

    Comparative proteomic investigation of Marsupenaeus japonicus hepatopancreas challenged with Vibrio parahaemolyticus and white spot syndrome virus

    Fish Shellfish Immunol.

    (2019)
  • L. Zhu et al.

    Differential proteome of haemocyte subpopulations responded to white spot syndrome virus infection in Chinese shrimp Fenneropenaeus chinensis

    Dev. Comp. Immunol.

    (2018)
  • M. Ning et al.

    Proteomic and metabolomic responses in hepatopancreas of whiteleg shrimp Litopenaeus vannamei infected by microsporidian Enterocytozoon hepatopenaei

    Fish Shellfish Immunol.

    (2019)
  • Z. Wang et al.

    Integrative microRNA and mRNA analysis reveals regulation of ER stress in the Pacific white shrimp Litopenaeus vannamei under acute cold stress

    Comp. Biochem. Physiol. Genom. Proteonomics

    (2020)
  • Y. Ma et al.

    Focal adhesion kinase regulates intestinal epithelial barrier function via redistribution of tight junction

    Biochim. Biophys. Acta (BBA) - Mol. Basis Dis.

    (2013)
  • O. Burgy et al.

    The WNT signaling pathways in wound healing and fibrosis

    Matrix Biol.

    (2018)
  • K. Hu et al.

    The roles of vascular endothelial growth factor in bone repair and regeneration

    Bone

    (2016)
  • D. Wu et al.

    Effects and transcriptional responses in the hepatopancreas of red claw crayfish Cherax quadricarinatus under cold stress

    J. Therm. Biol.

    (2019)
  • M.B. New

    A review of dietary studies with shrimp and prawns

    Aquaculture

    (1976)
  • P. Zhang et al.

    Swimming ability and physiological response to swimming fatigue in whiteleg shrimp, Litopenaeus vannamei

    Comp. Biochem. Physiol., A

    (2006)
  • Z. Wang et al.

    Physiological responses of pacific white shrimp Litopenaeus vannamei to temperature fluctuation in low-salinity water

    Front. Physiol.

    (2019)
  • J. Li et al.

    Schisandrin B prevents ulcerative colitis and colitis-associated-cancer by activating focal adhesion kinase and influence on gut microbiota in an in vivo and in vitro model

    Eur. J. Pharmacol.

    (2019)
  • M.R.I. Khan et al.

    Activation of focal adhesion kinase via M1 muscarinic acetylcholine receptor is required in restitution of intestinal barrier function after epithelial injury

    Biochim. Biophys. Acta (BBA) - Mol. Basis Dis.

    (2014)
  • H. Yang et al.

    Comparative transcriptome analysis of red swamp crayfish (Procambarus clarkia) hepatopancreas in response to WSSV and Aeromonas hydrophila infection

    Fish Shellfish Immunol.

    (2018)
  • M.H. Noureldein et al.

    Gut microbiota and mTOR signaling: insight on a new pathophysiological interaction

    Microb. Pathog.

    (2018)
  • P.H. Wang et al.

    Molecular cloning, characterization and expression analysis of two novel Tolls (LvToll2 and LvToll3) and three putative Spätzle-like Toll ligands (LvSpz1–3) from Litopenaeus vannamei

    Dev. Comp. Immunol.

    (2012)
  • P.H. Wang et al.

    An immune deficiency homolog from the white shrimp, Litopenaeus vannamei , activates antimicrobial peptide genes

    Mol. Immunol.

    (2009)
  • D. Tang et al.

    Comparative transcriptome analysis of the gills of Procambarus clarkii provides novel insights into the immune-related mechanism of copper stress tolerance

    Fish Shellfish Immunol.

    (2020)
  • J.A. Pérez-Romero et al.

    Impact of short-term extreme temperature events on physiological performance of Salicornia ramosissima J. Woods under optimal and sub-optimal saline conditions

    Sci Rep-Uk.

    (2019)
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