Proteomics reveals the gender differences in humoral immunity and physiological characteristics associated with reproduction in the sea cucumber Apostichopus japonicus
Graphical abstract
Introduction
In many invertebrates, gender and associated reproductive processes were proven to exert significant effects on immunity and physiological characteristics. For example, in the clam, Ruditapes philippinarum, acid phosphatase activity in hemocyte lysates from males was significantly higher than that from females [1]. Stronger mtDNA mutation and higher metabolic flexibility were observed in male and female Drosophila melanogaster, respectively [2]. In addition, locomotory performance was significantly depressed and lipid metabolism was substantially changed in the female sea cucumber, Apostichopus japonicus, following reproduction [3]. Immunity and physiology exert a crucial influence on survival and growth, respectively. Therefore, in the case of economically important invertebrates, gender and associated reproductive processes are important factors that need to be taken into account in improving breeding efficiency.
A. japonicus is one of the most important aquaculture species in East Asian countries, in terms of providing nutritional seafood as well as raw material for health care products and medicines [4]. Due to significant improvements in the quality of life of people in recent years, the market demand for A. japonicus has increased rapidly. As the culture zone of A. japonicus is limited, meticulous breeding has become an important method by which the culture efficiency of A. japonicus can be increased [5]. A. japonicus is a gonochoristic species that always experiences 2 reproductive processes while being cultured from juvenile to saleable adult stage [6]. Hence, gender and associated reproductive processes may exert an important effect on the breeding efficiency of A. japonicus. Until now, minor studies have been performed to explore immune differences between A. japonicus females and males. The gene expression profiles of the coelomocytes of A. japonicus females and males before and after spawning were determined using the transcriptome method [7], while the activities of a few immune-related enzymes in cell-free coelomic fluid (CCF) and coelomocytes of viripotent A. japonicus males and females were determined via biochemical methods [8]. Preliminary results have indicated a significant difference between the immunity of females and males. However, global gender differences in humoral immunity and physiological characteristics associated with reproduction in A. japonicus remain unclear; thereby restricting the development of appropriate culture strategies and methods based on gender and associated reproduction in the meticulous breeding of A. japonicus.
Rapid development of high-resolution mass spectrometry has enabled the systematic identification of proteins in the body fluids of animals as well as comparison of their expression levels [9,10]. Proteomics technology based on high-resolution mass spectrometry has become an effective tool for studying the responses of organisms to different changes [[11], [12], [13]]. Currently, it has been successfully applied to study immune or physiological responses of A. japonicus under various conditions, including body color [14], skin ulceration [15], heat shock [16], and pathogen challenges [17], as well the mechanisms underlying such responses. The current study compared overall protein differences in male and female A. japonicus CCF before and after spawning, using label-free proteomics technology, in order to explore the global gender differences in humoral immunity and physiological characteristics associated with A. japonicas reproduction. Our study was intended to provide essential data that would enable the development of strategies that would enhance meticulous breeding in A. japonicus.
Section snippets
Experimental animals
Sixty viripotent, A. japonicus sea cucumbers (body weight of 257 ± 18.6 g) were collected from Dalian, China in June 2018. These were acclimated in the laboratory for one week before use. Seawater used for temporary culture was filtered through sand and 300 μm nylon sieves. Seawater aquaria were maintained under conditions of 16 °C, a salinity level of 31‰, a pH of 8.2, and continuous aeration.
The collection of CCF and determination of sex
Before spawning, coelomic fluid was collected in a sterilized culture dish following dissection of
Influences of gender and reproduction on the protein expression in CCF
A total of 6839 peptides were obtained from 12 samples using label-free LC-MS/MS proteomic analyses. Following data filtering, a set of 1466 proteins were identified in the CCF of male and female A. japonicus before and after spawning. PLS-DA was applied to investigate the comprehensive divergence of protein expression profiles in different samples, which revealed that samples with the same gender and reproductive state were clustered together and away from others (Fig. 1). This result
Conclusions
A proteomic analysis based on label-free technology was performed to study the protein expression profiles in the CCF of male and female A. japonicus before and after spawning. The analysis revealed that both gender and reproduction significantly affected the composition and expression level of proteins in the CCF of A. japonicus. Our study found that the reproductive process exerted different but complicated gender based effects on the humoral immunity of A. japonicus. However, the alternative
Declaration of Competing Interest
The authors declare that there is no conflict of interests regarding the publication of this article.
Acknowledgements
This work was supported by grants from National Key R&D Program of China (2018YFD0900105), National Natural Science Foundation of China (31672688), Liaoning Revitalization Talents Program (XLYC1802091), Innovation, and Entrepreneurship Program for High-level Talent of Dalian (2018RQ60, 2018RD10) and Ocean & Fisheries Project of Liaoning Province (201823).
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Jingwei Jiang and Zelong Zhao are co-first authors.