Full length articleNew insights into the immune regulation and tissue repair of Litopenaeus vannamei during temperature fluctuation using TMT-based proteomics
Graphical abstract
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)
- et al.
Mortality risk attributable to high and low ambient temperature: a multicountry observational study
Lancet
(2015) - et al.
Cell cycle regulation and apoptosis mediated by p53 in response to hypoxia in hepatopancreas of the white shrimp Litopenaeus vannamei
Chemosphere
(2018) - et al.
Changes in the intestine barrier function of Litopenaeus vannamei in response to pH stress
Fish Shellfish Immunol.
(2019) - et al.
Response of gut microbiota to salinity change in two euryhaline aquatic animals with reverse salinity preference
Aquaculture
(2016) - et al.
Transcriptomic and microbiota response on Litopenaeus vannamei intestine subjected to acute sulfide exposure
Fish Shellfish Immunol.
(2019) - et al.
Effects of ammonia stress on the hemocytes of the Pacific white shrimp Litopenaeus vannamei
Chemosphere
(2020) - et al.
Comparative proteomic identification of the hepatopancreas response to cold stress in white shrimp
Litopenaeus vannamei, Aquaculture
(2016) - et al.
The immune defense response of Pacific white shrimp (Litopenaeus vannamei) to temperature fluctuation
Fish Shellfish Immunol.
(2020) - et al.
A new insight into the intestine of Pacific white shrimp: regulation of intestinal homeostasis and regeneration in Litopenaeus vannamei during temperature fluctuation
Comp. Biochem. Physiol. Genom. Proteonomics
(2020) - et al.
Detection of biomarkers to differentiate endocrine disruption from hepatotoxicity in zebrafish (Danio rerio) using proteomics
Chemosphere
(2020)