Acute environmental changes cause stress during conventional deep-sea biological sampling without in situ fixation and affect gene expressions of samples collected. However, the degree of influence and underlying mechanisms are hardly investigated. Here, we conducted comparative transcriptomic analyses between in situ and onboard fixed gills and between in situ and onboard fixed mantles of deep-sea mussel Gigantidas haimaensis to assess the effects of incidental sampling stress. Results showed that transcription, translation, and energy metabolism were upregulated in onboard fixed gills and mantles, thereby mobilizing rapid gene expression to tackle the stress. Autophagy and phagocytosis that related to symbiotic interactions between the host and endosymbiont were downregulated in the onboard fixed gills. These findings demonstrated that symbiotic gill and nonsymbiotic mantle responded differently to sampling stress, and symbiosis in the gill was perturbed. Further comparative metatranscriptomic analysis between in situ and onboard fixed gills revealed that stress response genes, peptidoglycan biosynthesis, and methane fixation were upregulated in the onboard fixed endosymbiotic Gammaproteobacteria inside the gills, implying that energy metabolism of the endosymbiont was increased to cope with sampling stress. Furthermore, comparative analysis between the mussel G. haimaensis and the limpet Bathyacmaea lactea transcriptomes resultedidentified six transcription factor orthologs upregulated in both onboard fixed mussel mantles and limpets, including sharply increased early growth response protein 1 and Kruppel-like factor 5. They potentially play key roles in initiating the response of sampled deep-sea macrobenthos to sampling stress. Our results clearly show that in situ fixed biological samples are vital for studying deep-sea environmental adaptation.
| [1] |
Bolger AM, Lohse M, Usadel B (2014). Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120.
|
| [2] |
Chen H, Wang M, Li M et al. (2021). A glimpse of deep-sea adaptation in chemosynthetic holobionts: Depressurization causes DNA fragmentation and cell death of methanotrophic endosymbionts rather than their deep-sea Bathymodiolinae host. Molecular Ecology 30, 2298–2312.
|
| [3] |
Chiu JJ, Wung BS, Hsieh HJ, Lo LW, Wang DL (1999). Nitric oxide regulates shear stress-induced early growth response-1. Expression via the extracellular signal-regulated kinase pathway in endothelial cells. Circulation Research 85, 238–246.
|
| [4] |
Chouchani ET, Kazak L, Spiegelman BM (2019). New advances in adaptive thermogenesis: UCP1 and beyond. Cell Metabolism 29, 27–37.
|
| [5] |
Davy SK, Allemand D, Weis VM (2012). Cell biology of cnidarian-dinoflagellate symbiosis. Microbiology and Molecular Biology Reviews 76, 229–261.
|
| [6] |
Ellis RJ, van der Vies SM (1991). Molecular chaperones. Annual Review of Biochemistry 60, 321–347.
|
| [7] |
Emms DM, Kelly S (2019). OrthoFinder: Phylogenetic orthology inference for comparative genomics. Genome Biology 20, 238.
|
| [8] |
Gomez-Pastor R, Burchfiel ET, Thiele DJ (2018). Regulation of heat shock transcription factors and their roles in physiology and disease. Nature Reviews Molecular Cell Biology 19, 4–19.
|
| [9] |
Grabherr MG, Haas BJ, Yassour M et al. (2011). Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nature Biotechnology 29, 644–652.
|
| [10] |
Hartl FU, Bracher A, Hayer-Hartl M (2011). Molecular chaperones in protein folding and proteostasis. Nature 475, 324–332.
|
| [11] |
He X, Xu T, Chen C et al. (2023). Same (sea) bed different dreams: Biological community structure of the Haima seep reveals distinct biogeographic affinities. The Innovation Geoscience 1, 100019.
|
| [12] |
Healy S, Khan P, Davie JR (2013). Immediate early response genes and cell transformation. Pharmacology & Therapeutics 137, 64–77.
|
| [13] |
Huang J, Huang P, Lu J et al. (2022). Gene expression profiles provide insights into the survival strategies in deep-sea mussel (Bathymodiolus platifrons) of different developmental stages. BMC Genomics 23, 311.
|
| [14] |
Kopylova E, Noé L, Touzet H (2012). SortMeRNA: Fast and accurate filtering of ribosomal RNAs in metatranscriptomic data. Bioinformatics 28, 3211–3217.
|
| [15] |
Li M, Gu Y, Ma Y-C et al. (2015). Kruppel-like factor 5 promotes epithelial proliferation and DNA damage repair in the intestine of irradiated mice. International Journal of Biological Sciences 11, 1458–1468.
|
| [16] |
Lin Y-T, Xu T, Chi-Ho Ip J et al. (2023). Interactions among deep-sea mussels and their epibiotic and endosymbiotic chemoautotrophic bacteria: Insights from multi-omics analysis. Zoological Research 44, 106–125.
|
| [17] |
McConnell BB, Ghaleb AM, Nandan MO, Yang VW (2007). The diverse functions of Kruppel-like factors 4 and 5 in epithelial biology and pathobiology. BioEssays 29, 549–557.
|
| [18] |
Motoki K, Watsuji T-O, Takaki Y, Takai K, Iwasaki W (2020). Metatranscriptomics by in situ RNA stabilization directly and comprehensively revealed episymbiotic microbial communities of deep-sea squat lobsters. mSystems 5, e00551-20.
|
| [19] |
Patro R, Duggal G, Love MI, Irizarry RA, Kingsford C (2017). Salmon provides fast and bias-aware quantification of transcript expression. Nature Methods 14, 417–419.
|
| [20] |
Perez M, Aroh O, Sun Y et al. (2023). Third-generation sequencing reveals the adaptive role of the epigenome in three deep-sea polychaetes. Molecular Biology and Evolution 40, msad172.
|
| [21] |
Ramakrishnan V (2002). Ribosome structure and the mechanism of translation. Cell 108, 557–572.
|
| [22] |
Robinson MD, Mccarthy DJ, Smyth GK (2010). edgeR: A bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139–140.
|
| [23] |
Semenza GL (2012). Hypoxia-inducible factors in physiology and medicine. Cell 148, 399–408.
|
| [24] |
Shen W-K, Chen S-Y, Gan Z-Q et al. (2023). AnimalTFDB 4.0: A comprehensive animal transcription factor database updated with variation and expression annotations. Nucleic Acids Research 51, D39–D45.
|
| [25] |
Sogin EM, Kleiner M, Borowski C, Gruber-Vodicka HR, Dubilier N (2021). Life in the dark: Phylogenetic and physiological diversity of chemosynthetic symbioses. Annual Review of Microbiology 75, 695–718.
|
| [26] |
Suess E (2020). Marine cold seeps: Background and recent advances. In: Wilkes H, ed. Hydrocarbons, Oils and Lipids: Diversity, Origin, Chemistry and Fate. Springer, Cham, pp. 747–767.
|
| [27] |
Sun X, Huang H, Pan X et al. (2019). EGR1 promotes the cartilage degeneration and hypertrophy by activating the Kruppel-like factor 5 and beta-catenin signaling. Biochimica et Biophysica Acta. Molecular Basis of Disease 1865, 2490–2503.
|
| [28] |
Sun J, Zhang Y, Xu T et al. (2017). Adaptation to deep-sea chemosynthetic environments as revealed by mussel genomes. Nature Ecology & Evolution 1, https://doi.org/10.1038/s41559-017-0121
|
| [29] |
Suzuki T, Sawaki D, Aizawa K et al. (2009). Kruppel-like Factor 5 shows proliferation-specific roles in vascular remodeling, direct stimulation of cell growth, and inhibition of apoptosis. Journal of Biological Chemistry 284, 9549–9557.
|
| [30] |
Tame A, Maruyama T, Ikuta T et al. (2023). mTORC1 regulates phagosome digestion of symbiotic bacteria for intracellular nutritional symbiosis in a deep-sea mussel. Science Advances 9, eadg8364.
|
| [31] |
Tietjen M (2020). Physiology and ecology of deep-sea Bathymodiolus symbioses (Dissertation). Universität Bremen, Bremen.
|
| [32] |
Voth W, Jakob U (2017). Stress-activated chaperones: A first line of defense. Trends Biochemical Science 42, 899–913.
|
| [33] |
Wahl MC, Will CL, Lührmann R (2009). The spliceosome: Design principles of a dynamic RNP machine. Cell 136, 701–718.
|
| [34] |
Wang H, Zhang H, Wang M, Chen H, Lian C, Li C (2019). Comparative transcriptomic analysis illuminates the host-symbiont interactions in the deep-sea mussel Bathymodiolus platifrons. Deep Sea Research Part I: Oceanographic Research Papers 151, 103082.
|
| [35] |
Xie Y, Li Y, Chen J, Ding H, Zhang X (2023). Early growth response-1: Key mediators of cell death and novel targets for cardiovascular disease therapy. Frontiers in Cardiovascular Medicine 10, 1162662.
|
| [36] |
Xu T, Feng D, Tao J, Qiu J-W (2019). A new species of deep-sea mussel (Bivalvia: Mytilidae: Gigantidas) from the South China Sea: Morphology, phylogenetic position, and gill-associated microbes. Deep-Sea Research Part I-Oceanographic Research Papers 146, 79–90.
|
| [37] |
Yan G, Lan Y, Sun J, Xu T, Wei T, Qian P-Y (2022). Comparative transcriptomic analysis of in situ and onboard fixed deep-sea limpets reveals sample preparation-related differences. Iscience 25, 104092.
|
| [38] |
Yan S-F, Lu J, Zou YS et al. (1999). Hypoxia-associated induction of early growth response-1 gene expression. Journal of Biological Chemistry 274, 15030–15040.
|
| [39] |
Yoo H, Bard JAM, Pilipenko EV, Drummond DA (2022). Chaperones directly and efficiently disperse stress-triggered biomolecular condensates. Molecular Cell 82, 741–755.e11.
|
| [40] |
Zheng P, Wang M, Li C et al. (2017). Insights into deep-sea adaptations and host–symbiont interactions: A comparative transcriptome study on Bathymodiolus mussels and their coastal relatives. Molecular Ecology 26, 5133–5148. Portico.
|
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2024 The Author(s). Integrative Zoology published by International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.