Transcriptome Analysis of the Central Nervous System of Sea Slug (Onchidium reevesii) Exposed to Low-Frequency Noise

Low-frequency noise has become a marine pollutant that cannot be ignored, but most studies have focused on the behavioral and physiological effects on marine vertebrates, with few studies in marine mollusks. Therefore, sea slug was used in this study to investigate the effect of low-frequency noise on its physiological aspects. This experiment was designed with different low-frequency noise (0, 100, 300, and 500 Hz) and different stimulation times (0, 6, and 12 h) to measure superoxide dismutase (SOD), malondialdehyde (MDA), and catalase (CAT) activities in hemolymph and transcriptomics in the control (C) and 6 and 12 h groups (L1 and L2) with 500 Hz noise. The results showed a positive correlation between antioxidant enzyme activity and low-frequency noise frequency (P < 0.05) and no correlation with time (P > 0.05). In central nervous system (CNS) transcriptomics, 2,460 and 3,268 genes had upregulated expression and 2,765 and 2,783 genes had downregulated expression in the L1 and L2 groups, respectively, compared to the C group. According to Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, low-frequency noise mainly affects signaling pathways such as cytokine-cytokine receptor interaction, the FoxO signaling pathway, natural killer cell-mediated cytotoxicity, apoptosis immune-related pathways, and energy metabolic pathways such as glycolysis, the TCA cycle, and glycerophospholipid metabolism, as well as neurological pathways such as GABAergic synapses, the synaptic vesicle cycle, amyotrophic lateral sclerosis (ALS) and other neurological pathways. This study would provide valuable reference information on the potential response of mollusks to low-frequency noise stress.

Ultraviolet screening by slug tissue and tight packing of plastids protect photosynthetic sea slugs from photoinhibition

One of the main mysteries regarding photosynthetic sea slugs is how the slug plastids handle photoinhibition, the constant light-induced damage to Photosystem II of photosynthesis. Recovery from photoinhibition involves proteins encoded by both the nuclear and plastid genomes, and slugs with plastids isolated from the algal nucleus are therefore expected to be incapable of constantly repairing the damage as the plastids inside the slugs grow old. We studied photoinhibition-related properties of the sea slug Elysia timida that ingests its plastids from the green alga Acetabularia acetabulum. Spectral analysis of both the slugs and the algae revealed that there are two ways the slugs use to avoid major photoinhibition of their plastids. Firstly, highly photoinhibitory UV radiation is screened by the slug tissue or mucus before it reaches the plastids. Secondly, the slugs pack the plastids tightly in their thick bodies, and therefore plastids in the outer layers protect the inner ones from photoinhibition. Both properties are expected to greatly improve the longevity of the plastids inside the slugs, as the plastids do not need to repair excessive amounts of damage.

Pigment and Fatty Acid Heterogeneity in the Sea Slug Elysia crispata Is Not Shaped by Habitat Depth

Long-term retention of functional chloroplasts in animal cells occurs only in sacoglossan sea slugs. Analysis of molecules related to the maintenance of these organelles can provide valuable information on this trait (kleptoplasty). The goal of our research was to characterize the pigment and fatty acid (FA) composition of the sea slug Elysia crispata and their associated chloroplasts that are kept functional for a long time, and to quantify total lipid, glycolipid and phospholipid contents, identifying differences between habitats: shallow (0–4 m) and deeper (8–12 m) waters. Specimens were sampled and analyzed after a month of food deprivation, through HPLC, GC-MS and colorimetric methods, to ensure an assessment of long-term kleptoplasty in relation to depth. Pigment signatures indicate that individuals retain chloroplasts from different macroalgal sources. FA classes, phospholipid and glycolipid contents displayed dissimilarities between depths. However, heterogeneities in pigment and FA profiles, as well as total lipid, glycolipid and phospholipid amounts in E. crispata were not related to habitat depth. The high content of chloroplast origin molecules, such as Chl a and glycolipids after a month of starvation, confirms that E. crispata retains chloroplasts in good biochemical condition. This characterization fills a knowledge gap of an animal model commonly employed to study kleptoplasty.

Photosynthesis from stolen chloroplasts can support sea slug reproductive fitness

Some sea slugs are able to steal functional chloroplasts (kleptoplasts) from their algal food sources, but the role and relevance of photosynthesis to the animal host remain controversial. While some researchers claim that kleptoplasts are slowly digestible ‘snacks’, others advocate that they enhance the overall fitness of sea slugs much more profoundly. Our analysis shows light-dependent incorporation of 13 C and 15 N in the albumen gland and gonadal follicles of the sea slug Elysia timida , representing translocation of photosynthates to kleptoplast-free reproductive organs. Long-chain polyunsaturated fatty acids with reported roles in reproduction were produced in the sea slug cells using labelled precursors translocated from the kleptoplasts. Finally, we report reduced fecundity of E. timida by limiting kleptoplast photosynthesis. The present study indicates that photosynthesis enhances the reproductive fitness of kleptoplast-bearing sea slugs, confirming the biological relevance of this remarkable association between a metazoan and an algal-derived organelle.

Ultraviolet screening by slug tissue and tight packing of plastids protect photosynthetic sea slugs from photoinhibition

One of the main unsolved questions regarding photosynthetic sea slugs is how the slug plastids handle photoinhibition of Photosystem II. Photoinhibition has not been studied in detail in these animals although resilience against photoinhibition might obviously explain the longevity of plastids inside animal cytosol. Light response and action spectrum of photoinhibition were measured from the slug Elysia timida and its prey alga Acetabularia acetabulum. Plastid packing in the slugs and algae was compared with spectroscopic and microscopic methods. The importance of plastid concentration was also estimated by measuring photoinhibition from starved slugs. Compared to A. acetabulum, E. timida is highly resistant against photoinhibition. The resilience of the slugs is even more pronounced in the UV-region, as the slug tissue screens UV radiation. The plastids in the slug tissue are tightly packed, and the outer plastids protect the inner ones from photoinhibition. The sea slug E. timida protects its plastids from photoinhibition by screening UV radiation and packing the plastids tightly in its tissues. Both mechanisms enhance the longevity of the plastids in slug cytosol and ameliorate the need for repair of photoinhibited Photosystem II.