Subcellular Partitioning Research Articles

Zn Subcellular Distribution in Liver of Goldfish (Carassius Auratus) with Exposure to Zinc Oxide Nanoparticles and Mechanism of Hepatic Detoxification

Zinc Oxide Nanoparticles (ZnO NPs) have attracted increasing concerns because of their widespread use and toxic potential. In this study, Zn accumulations in different tissues (gills, liver, muscle, and gut) of goldfish (Carassius auratus) after exposure to ZnO NPs were studied in comparison with bulk ZnO and Zn2+. And the technique of subcellular partitioning was firstly used on the liver of goldfish to study the hepatic accumulation of ZnO NPs. The results showed that at sublethal Zn concentration (2 mg/L), bioaccumulation in goldfish was tissue-specific and dependent on the exposure materials. Compared with Zn2+, the particles of bulk ZnO and the ZnO NPs appeared to aggregate in the environmentally contacted tissues (gills and gut), rather than transport to the internal tissues (liver and muscle). The subcellular distributions of liver differed for the three exposure treatments. After ZnO NPs exposure, Zn percentage in metal-rich granule (MRG) increased significantly, and after Zn2+ exposure, it increased significantly in the organelles. Metallothionein-like proteins (MTLP) were the main target for Zn2+, while MRG played dominant role for ZnO NPs. The different results of subcellular distributions revealed that metal detoxification mechanisms of liver for ZnO NPs, bulk ZnO, and Zn2+ were different. Overall, subcellular partitioning provided an interesting start to better understanding of the toxicity of nano- and conventional materials.

Uptake and subcellular partitioning of trivalent metals in a green alga: comparison between Al and Sc

Despite 40+ years of research on aluminum (Al) toxicity in aquatic organisms, Al transport mechanisms through biological membranes, and the intracellular fate of Al once assimilated, remain poorly understood. The trivalent metal scandium shares chemical similarities with Al and, unlike Al, it has a convenient radioactive tracer (Sc-46) allowing for relatively simple measurements at environmentally relevant concentrations. Thus, we investigated the potential of Sc to substitute for Al in uptake and intracellular fate studies with the green alga Chlamydomonas reinhardtii. Short-term (<60 min) competitive uptake experiments indicated that Al does not inhibit Sc influx, implying that these metals do not share a common transport mechanism. Also, internalized Al concentrations were ~4 times higher than Sc concentrations after long-term (72 h) exposures under similar conditions (4.5 μM AlT or ScT, 380 μM FT, pH 7.0, 3.8 pM Al calc (3+) and 1.0 pM Sc calc (3+) ). However, interesting similarities were observed in their relative subcellular distributions, suggesting possible common toxicity/tolerance mechanisms. Both metals mostly distributed to the organelles fraction and almost no association was found with the cytosolic proteins. The greatest difference was observed in the cellular debris fraction (membranes and nucleus) where Al was much more concentrated than Sc. However, it is not clear whether or not this fraction contained extracellular metal associated with the algal surface. To summarize, Sc does not seem to be an adequate substitute of Al for transport/uptake studies, but could be for investigations of toxicity/tolerance mechanisms in C. reinhardtii. Further work is needed to verify this latter suggestion.

Cephalopods as vectors of harmful algal bloom toxins in marine food webs.

Here we summarize the current knowledge on the transfer and accumulation of harmful algal bloom (HAB)-related toxins in cephalopods (octopods, cuttlefishes and squids). These mollusks have been reported to accumulate several HAB-toxins, namely domoic acid (DA, and its isomers), saxitoxin (and its derivatives) and palytoxin (and palytoxin-like compounds) and, therefore, act as HAB-toxin vectors in marine food webs. Coastal octopods and cuttlefishes store considerably high levels of DA (amnesic shellfish toxin) in several tissues, but mainly in the digestive gland (DG)—the primary site of digestive absorption and intracellular digestion. Studies on the sub-cellular partitioning of DA in the soluble and insoluble fractions showed that nearly all DA (92.6%) is found in the cytosol. This favors the trophic transfer of the toxins since cytosolic substances can be absorbed by predators with greater efficiency. The available information on the accumulation and tissue distribution of DA in squids (e.g., in stranded Humboldt squids, Dosidicus gigas) is scarcer than in other cephalopod groups. Regarding paralytic shellfish toxins (PSTs), these organisms accumulate them at the greatest extent in DG >> kidneys > stomach > branchial hearts > posterior salivary glands > gills. Palytoxins are among the most toxic molecules identified and stranded octopods revealed high contamination levels, with ovatoxin (a palytoxin analogue) reaching 971 μg kg−1 and palytoxin reaching 115 μg kg−1 (the regulatory limit for PlTXs is 30 μg kg−1 in shellfish). Although the impacts of HAB-toxins in cephalopod physiology are not as well understood as in fish species, similar effects are expected since they possess a complex nervous system and highly developed brain comparable to that of the vertebrates. Compared to bivalves, cephalopods represent a lower risk of shellfish poisoning in humans, since they are usually consumed eviscerated, with exception of traditional dishes from the Mediterranean area.

Inter-site differences of zinc susceptibility of the oyster Crassostrea hongkongensis

Understanding the underlying mechanisms governing metal toxicity is crucial for predicting the risks and effects of metal pollutants. We hypothesized that metal toxicity is related to a threshold concentration of metabolically available metal but not to the total body metal concentration. Following a two-month laboratory Zn exposure, we characterized mortality and Zn bioaccumulation and subcellular partitioning in the oyster Crassostrea hongkongensis sampled from three sites with contrasting histories of Zn exposure and one multiple-metal contaminated site. Large differences in Zn sensitivity, lethal body concentration, and detoxification capability between sites were observed. Specifically, the oysters from the highly Zn-contaminated site were more tolerant to Zn exposure than those from the relatively clean ones, and the former accumulated and detoxified more Zn and had a significantly higher lethal body Zn concentration. The accumulation of Zn in the metabolically available pool (operationally defined as the metal-sensitive fraction) in the oysters from the multiple-metal contaminated site was relatively fast, and correspondingly they were highly sensitive to Zn exposure. The lethal threshold concentration of total body Zn varied significantly within the four sites, and thus total body Zn concentration could not serve as a suitable toxicity indicator. Importantly, Zn accumulation within the operationally defined metabolically available pool better explained variances in mortality than Zn accumulation in the whole body. Our results suggested that Zn toxicity is governed by its accumulation in the metabolically available pool, not the total accumulated Zn concentration.

Influence of a step-change in metal exposure (Cd, Cu, Zn) on metal accumulation and subcellular partitioning in a freshwater bivalve, Pyganodon grandis: A long-term transplantation experiment between lakes with contrasting ambient metal levels

The objective of the present field experiment was to identify detoxification responses in the gills and digestive gland of a freshwater unionid bivalve, Pyganodon grandis, subjected to a step-change in metal exposure. Adult bivalves were transferred from a reference site (Lake Opasatica) and a metal-contaminated lake (Lake Héva) to a second contaminated lake (Lake Vaudray) in northwestern Quebec, Canada. Changes in organ metal concentrations, in the subcellular distribution of metals and in metallothionein concentrations were followed over time (t=0, 132, (400) and 860 days). At each collection time and for each bivalve, the gills and digestive gland were excised and gently homogenized; six sub-cellular fractions were separated by differential centrifugation and analyzed for their Cd, Cu and Zn content, and metallothionein was quantified independently.Metal detoxification strategies were shown to differ between target organs: in the gills, incoming metals were sequestered largely in the granules, whereas in the digestive gland the same metals primarily accumulated in the cytosol, in the metallothionein-like protein fraction. These metal-handling strategies, as employed by the metal-naïve bivalves originating in the reference lake, closely resemble those identified in free-living P. grandis chronically exposed in the metal-contaminated lake, suggesting that the ability to handle incoming metals (Cd in particular) is inherent in P. grandis and is not a trait acquired after long-term adaptation of the bivalve to metal-contaminated environments.The bivalves transplanted from both Lakes Opasatica and Héva were able to tolerate their new surroundings during the first 400 days of the transplant experiment, as indicated by the absence of mortality and the presence of gravid animals. Over the final 460 days, mortality remained low for the bivalves transplanted from the reference lake (20%) but reached 100% in the transplanted group from the contaminated lake. It would seem that the Lake Héva bivalves were compromised by their initial exposure to metals in their home lake and that the added stress of being transplanted to and caged in a lake with comparable or slightly higher concentrations of metals was sufficient to cause mortality.

Subcellular fractionation and chemical speciation of uranium to elucidate its fate in gills and hepatopancreas of crayfish Procambarus clarkii

Knowledge of the organ and subcellular distribution of metals in organisms is fundamental for the understanding of their uptake, storage, elimination and toxicity. Detoxification via MTLP and MRG formation and chelation by some proteins are necessary to better assess the metal toxic fraction in aquatic organisms. This work focused on uranium, natural element mainly used in nuclear industry, and its subcellular fractionation and chemical speciation to elucidate its accumulation pattern in gills and hepatopancreas of crayfish Procambarus clarkii, key organs of uptake and detoxification, respectively. Crayfish waterborne exposure was performed during 4 and 10d at 0, 30, 600 and 4000μgUL−1. After tissue dissection, uranium subcellular fractionation was performed by successive ultracentrifugations. SEC-ICP MS was used to study uranium speciation in cytosolic fraction. The uranium subcellular partitioning patterns varied according to the target organ studied and its biological function in the organism. The cytosolic fraction accounted for 13–30% of the total uranium amount in gills and 35–75% in hepatopancreas. The uranium fraction coeluting with MTLPs in gills and hepatopancreas cytosols showed that roughly 55% of uranium remained non-detoxified and thus potentially toxic in the cytosol. Furthermore, the sum of uranium amount in organelle fractions and in the non-detoxified part of cytosol, possibly equivalent to available fraction, accounted for 20% (gills) and 57% (hepatopancreas) of the total uranium. Finally, the SEC-ICP MS analysis provided information on potential competition of U for biomolecules similar than the ones involved in endogenous essential metal (Fe, Cu) chelation.

Examining strategies to facilitate vitamin B1 biofortification of plants by genetic engineering

Thiamin (vitamin B1) is made by plants and microorganisms but is an essential micronutrient in the human diet. All organisms require it as a cofactor in its form as thiamin pyrophosphate (TPP) for the activity of key enzymes of central metabolism. In humans, deficiency is widespread particularly in populations where polished rice is a major component of the diet. Considerable progress has been made on the elucidation of the biosynthesis pathway within the last few years enabling concrete strategies for biofortification purposes to be devised, with a particular focus here on genetic engineering. Furthermore, the vitamin has been shown to play a role in both abiotic and biotic stress responses. The precursors for de novo biosynthesis of thiamin differ between microorganisms and plants. Bacteria use intermediates derived from purine and isoprenoid biosynthesis, whereas the pathway in yeast involves the use of compounds from the vitamin B3 and B6 groups. Plants on the other hand use a combination of the bacterial and yeast pathways and there is subcellular partitioning of the biosynthesis steps. Specifically, thiamin biosynthesis occurs in the chloroplast of plants through the separate formation of the pyrimidine and thiazole moieties, which are then coupled to form thiamin monophosphate (TMP). Phosphorylation of thiamin to form TPP occurs in the cytosol. Therefore, thiamin (or TMP) must be exported from the chloroplast to the cytosol for the latter step to be executed. The regulation of biosynthesis is mediated through riboswitches, where binding of the product TPP to the pre-mRNA of a biosynthetic gene modulates expression. Here we examine and hypothesize on genetic engineering approaches attempting to increase the thiamin content employing knowledge gained with the model plant Arabidopsis thaliana. We will discuss the regulatory steps that need to be taken into consideration and can be used a prerequisite for devising such strategies in crop plants.

Transformation, Localization, and Biomolecular Binding of Hg Species at Subcellular Level in Methylating and Nonmethylating Sulfate-Reducing Bacteria

Microbial activity is recognized to play an important role on Hg methylation in aquatic ecosystems. However, the mechanism at the cellular level is still poorly understood. In this work subcellular partitioning and transformation of Hg species in two strains: Desulfovibrio sp. BerOc1 and Desulfovibrio desulfuricans G200 (which exhibit different Hg methylation potential) are studied as an approach to the elucidation of Hg methylation/demethylation processes. The incubation with isotopically labeled Hg species ((199)Hgi and Me(201)Hg) not only allows the determination of methylation and demethylation rates simultaneously, but also the comparison of the localization of the originally added and resulting species of such metabolic processes. A dissimilar Hg species distribution was observed. In general terms, monomethylmercury (MeHg) is preferentially localized in the extracellular fraction; meanwhile inorganic mercury (Hgi) is associated to the cells. The investigation of Hg binding biomolecules on the cytoplasmatic and extracellular fractions (size exclusion chromatography coupled to ICP-MS) revealed noticeable differences in the pattern corresponding to the Hg methylating and nonmethylating strains.

Down-regulation of the TaGW2 gene by RNA interference results in decreased grain size and weight in wheat

For important food crops such as wheat and rice, grain yield depends on grain number and size. In rice (Oryza sativa), GW2 was isolated from a major quantitative trait locus for yield and encodes an E3 RING ligase that negatively regulates grain size. Wheat (Triticum aestivum) has TaGW2 homologues in the A, B, and D genomes, and polymorphisms in TaGW2-A were associated with grain width. Here, to investigate TaGW2 function, RNA interference (RNAi) was used to down-regulate TaGW2 transcript levels. Transgenic wheat lines showed significantly decreased grain size-related dimensions compared with controls. Furthermore, TaGW2 knockdown also caused a significant reduction in endosperm cell number. These results indicate that TaGW2 regulates grain size in wheat, possibly by controlling endosperm cell number. Wheat and rice GW2 genes thus seem to have divergent functions, with rice GW2 negatively regulating grain size and TaGW2 positively regulating grain size. Analysis of transcription of TaGW2 homoeologues in developing grains suggested that TaGW2-A and -D act in both the division and late grain-filling phases. Furthermore, biochemical and molecular analyses revealed that TaGW2-A is a functional E3 RING ubiquitin ligase with nucleocytoplasmic subcellular partitioning. A functional nuclear export sequence responsible for TaGW2-A export from the nucleus to the cytosol and retention in the nucleolus was identified. Therefore, these results show that TaGW2 acts in the regulation of grain size and may provide an important tool for enhancement of grain yield.

Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate

Bioluminescence methodologies have been extraordinarily useful due to their high sensitivity, broad dynamic range, and operational simplicity. These capabilities have been realized largely through incremental adaptations of native enzymes and substrates, originating from luminous organisms of diverse evolutionary lineages. We engineered both an enzyme and substrate in combination to create a novel bioluminescence system capable of more efficient light emission with superior biochemical and physical characteristics. Using a small luciferase subunit (19 kDa) from the deep sea shrimp Oplophorus gracilirostris, we have improved luminescence expression in mammalian cells ∼2.5 million-fold by merging optimization of protein structure with development of a novel imidazopyrazinone substrate (furimazine). The new luciferase, NanoLuc, produces glow-type luminescence (signal half-life >2 h) with a specific activity ∼150-fold greater than that of either firefly (Photinus pyralis) or Renilla luciferases similarly configured for glow-type assays. In mammalian cells, NanoLuc shows no evidence of post-translational modifications or subcellular partitioning. The enzyme exhibits high physical stability, retaining activity with incubation up to 55 °C or in culture medium for >15 h at 37 °C. As a genetic reporter, NanoLuc may be configured for high sensitivity or for response dynamics by appending a degradation sequence to reduce intracellular accumulation. Appending a signal sequence allows NanoLuc to be exported to the culture medium, where reporter expression can be measured without cell lysis. Fusion onto other proteins allows luminescent assays of their metabolism or localization within cells. Reporter quantitation is achievable even at very low expression levels to facilitate more reliable coupling with endogenous cellular processes.

Prediction of metal toxicity in aquatic organisms

Metal pollution has been a major environmental problem in China with the increasing industrialization. The prediction of metal toxicity is extremely challenging due to the complex metal handling and sequestration strategies of different aquatic organisms. In this review, the recent progress made in this area is discussed. In particular, the subcellular partitioning model which has gained recognition in recent years is highlighted. The subcellular partitioning model appears to be dependable for predicting the toxicity in unicellular phytoplankton. It is important to understand the differential ways that metals bind to different subcellular pools and their ecotoxicological significance in aquatic organisms under different exposure regimes. It is also critical to appreciate that every metal is unique to each aquatic species. Despite the huge progress made over the past 30 years, much remains to be done to fully understand metal toxicity in aquatic organisms.

Selective loss of RPGRIP1-dependent ciliary targeting of NPHP4, RPGR and SDCCAG8 underlies the degeneration of photoreceptor neurons

The retinitis pigmentosa GTPase regulator (RPGR) and nephrocystin-4 (NPHP4) comprise two key partners of the assembly complex of the RPGR-interacting protein 1 (RPGRIP1). Mutations in RPGR and NPHP4 are linked to severe multisystemic diseases with strong retinal involvement of photoreceptor neurons, whereas those in RPGRIP1 cause the fulminant photoreceptor dystrophy, Leber congenital amaurosis (LCA). Further, mutations in Rpgrip1 and Nphp4 suppress the elaboration of the outer segment compartment of photoreceptor neurons by elusive mechanisms, the understanding of which has critical implications in uncovering the pathogenesis of syndromic retinal dystrophies. Here we show RPGRIP1 localizes to the photoreceptor connecting cilium (CC) distally to the centriole/basal body marker, centrin-2 and the ciliary marker, acetylated-α-tubulin. NPHP4 abuts proximally RPGRIP1, RPGR and the serologically defined colon cancer antigen-8 (SDCCAG8), a protein thought to partake in the RPGRIP1 interactome and implicated also in retinal–renal ciliopathies. Ultrastructurally, RPGRIP1 localizes exclusively throughout the photoreceptor CC and Rpgrip1nmf247 photoreceptors present shorter cilia with a ruffled membrane. Strikingly, Rpgrip1nmf247 mice without RPGRIP1 expression lack NPHP4 and RPGR in photoreceptor cilia, whereas the SDCCAG8 and acetylated-α-tubulin ciliary localizations are strongly decreased, even though the NPHP4 and SDCCAG8 expression levels are unaffected and those of acetylated-α-tubulin and γ-tubulin are upregulated. Further, RPGRIP1 loss in photoreceptors shifts the subcellular partitioning of SDCCAG8 and NPHP4 to the membrane fraction associated to the endoplasmic reticulum. Conversely, the ciliary localization of these proteins is unaffected in glomeruli or tubular kidney cells of Rpgrip1nmf247, but NPHP4 is downregulated developmentally and selectively in kidney cortex. Hence, RPGRIP1 presents cell type-dependent pathological effects crucial to the ciliary targeting and subcellular partitioning of NPHP4, RPGR and SDCCAG8, and acetylation of ciliary α-tubulin or its ciliary targeting, selectively in photoreceptors, but not kidney cells, and these pathological effects underlie photoreceptor degeneration and LCA.

Endothelial Caveolar Hub Regulation of Adenosine Triphosphate–Induced Endothelial Nitric Oxide Synthase Subcellular Partitioning and Domain-Specific Phosphorylation

ATP leads to endothelial NO synthase (eNOS)/NO-mediated vasodilation, a process hypothesized to depend on the endothelial caveolar eNOS partitioning and subcellular domain-specific multisite phosphorylation state. We demonstrate herein that, in both the absence and presence of ATP, the uterine artery endothelial caveolae contain specific protein machinery related to subcellular partitioning and act as specific focal "hubs" for NO- and ATP-related proteins. ATP-induced eNOS regulation showed a complex set of multisite posttranslational phosphorylation events that were closely associated with the enzyme's partitioning between caveolar and noncaveolar endothelial subcellular domains. The comprehensive model that we present demonstrates that ATP repartitioned eNOS between the caveolar and noncaveolar subcellular domains; specifically, the stimulatory (PSer635)eNOS was substantially higher in the caveolar pool with subcellular domain-independent increased levels on ATP treatment. The stimulatory (PSer1179)eNOS was not altered by ATP treatment. However, the inhibitory (PThr495)eNOS was regulated predominantly in the caveolar domain with decreased levels on ATP action. In contrast, the agonist-specific (PSer114)eNOS was localized in the noncaveolar pool with increased levels on ATP stimulation. Thus, the endothelial caveolar membrane system plays a pivotal role(s) in ATP-associated subcellular partitioning and possesses the relevant protein machinery for ATP-induced NO regulation. Furthermore, these subcellular domain-specific phosphorylation/dephosphorylation events provide evidence relating to eNOS spatio-temporal dynamics.

Subcellular partitioning of elements and availability for trophic transfer: Comparison between the Bivalve Cerastoderma edule and the Polychaete Diopatra neapolitana

Metal transference through food-chain can constitute a serious problem, particularly in coastal systems, where macrobenthic organisms are often exposed to metal contamination. Previous studies have shown that the elements accumulated in macrobenthic organisms can either be precipitated or soluble in the cells. Whilst differences in the subcellular distribution between fractions are described for Cd, Cu and Zn, little is known for other elements such as As, Pb or Hg, despite their toxicity. The present study compared the subcellular partitioning of several elements in two different macrobenthic species, the bivalve Cerastoderma edule and the polychaete Diopatra neapolitana, and the potential that each element has to be trophically available was determined. There were differences in element accumulation between the two species: D. neapolitana presented a higher concentration of Zn and As while, C. edule accumulated more Al, Ni and Pb. The results obtained for the soluble fraction showed that the elements in higher concentration were, for both species, Al and Zn. In addition to these two elements, Cu, Cd, Cr and As concentrations in this fraction were higher in D. neapolitana than in C. edule, although the bivalve presented higher Ni levels. These results give a good estimate of the amount of elements more readily available to be trophically transferred. The relative amount of metal present in the soluble fraction may indicate more clearly the ability of each species to cope with each element and thus their propensity to allocate subcellular elements, regardless of the environment that it inhabits. The percentage of elements in the soluble fraction showed that C. edule had a high predisposition to compartmentalise As and Cd in the trophically available fraction, given that the other elements were preferentially in the insoluble fraction. In contrast, D. neapolitana distributes metals and As more evenly throughout both fractions. As our study simultaneously evaluated the different elements, this gave a more precise idea of the real contamination that passes to higher trophic levels, especially in the elements that have high toxicity and which had not yet been evaluated (Hg, Pb, and As). This shows that contamination of the food webs of an ecosystem is not only dependent on its chemical contamination, but also on the organisms at each trophic level.

Subcellular partitioning links BLM‐based toxicokinetics for assessing cadmium toxicity to rainbow trout

The purpose of this article was to develop an integrated-scale toxicological model to investigate the impact of cadmium (Cd) toxicity on rainbow trout (Oncorhynchus mykiss) based on recent published experimental data. This model was generated from three different types of functional relationship: biotic ligand model (BLM), damage assessment model (DAM), and subcellular partitioning model (SPM), both of key toxicological determinants involved and of functional connections between them. Toxicokinetic parameters of uptake rate constant (k(1)) and elimination rate constant (k(2)) in gill, liver, and subcellular fractions were derived. A negative correlation between gill binding fraction of Cd and bioaccumulation factor was found. Detoxifying ability (% detoxified in liver metabolically detoxified pool (MDP)) and k(2) were negatively correlated, indicating that increasing % detoxified in MDP can compensate for lower k(2). This finding suggests a potential tradeoff between the abilities of elimination and detoxification for Cd. Yet, compensation between the ability to eliminate Cd and the ability to recover Cd-induced damage was not found. However, changes in k(2) and recovery rate constant (k(r)) can shift the dynamics of Cd susceptibility probability. This analysis implicates that once k(2) is determined experimentally, the values of k(r) and % detoxified in MDP can be predicted by the proposed k(2)-k(r) and k(2)-% detoxified relationships. This study suggests that the mechanistic linking of BLM-based DAM and SPM can incorporate the organ- and cell-scale exposure experimental data to investigate the mechanisms of ecophysiological response for aquatic organisms exposed to metal stressors.

Parasites modify sub-cellular partitioning of metals in the gut of fish

Infestation of fish by parasites may influence metal accumulation patterns in the host. However, the subcellular mechanisms of these processes have rarely been studied. Therefore, this study determined how a cyprinid fish ( Rastrineobola argentea) partitioned four metals (Cd, Cr, Zn and Cu) in the subcellular fractions of the gut in presence of an endoparasite ( Ligula intestinalis). The fish were sampled along four sites in Lake Victoria, Kenya differing in metal contamination. Accumulation of Cd, Cr and Zn was higher in the whole body and in the gut of parasitized fish compared to non-parasitized fish, while Cu was depleted in parasitized fish. Generally, for both non-parasitized and parasitized fish, Cd, Cr and Zn partitioned in the cytosolic fractions and Cu in the particulate fraction. Metal concentrations in organelles within the particulate fractions of the non-parasitized fish were statistically similar except for Cd in the lysosome, while in the parasitized fish, Cd, Cr and Zn were accumulated more by the lysosome and microsomes. In the cytosolic fractions, the non-parasitized fish accumulated Cd, Cr and Zn in the heat stable proteins (HSP), while in the parasitized fish the metals were accumulated in the heat denatured proteins (HDP). On the contrary, Cu accumulated in the HSP in parasitized fish. The present study revealed specific binding of metals to potentially sensitive sub-cellular fractions in fish in the presence of parasites, suggesting interference with metal detoxification, and potentially affecting the health status of fish hosts in Lake Victoria.

Trade-offs between elimination and detoxification in rainbow trout and common bivalve molluscs exposed to metal stressors

The purpose of this paper was to examine trade-offs between elimination and detoxification in rainbow trout and three common bivalve molluscs (clam, oyster, and scallop) exposed to cadmium (Cd), copper (Cu), and zinc (Zn) based on recent reported experimental data. We incorporated metal influx threshold with subcellular partitioning to estimate rate constants of detoxification ( k d) and elimination ( k 2). We found that the relationships between k 2 and k d were negative for rainbow trout and positive for bivalve molluscs. However, the relationships between k d and % metal in metabolically detoxified pool were found positive for rainbow trout and negative for bivalve molluscs. Our results also indicated that rainbow trout had higher accumulation (∼60–90%) in metabolically active pool when exposed to essential metals of Cu and Zn and had only 10–50% accumulation in response to non-essential metal of Cd. Based on a cluster analysis, this study indicated that similarity of physiological regulations among study species was found between Cd and Zn. Our study suggested that detoxification can be predicted by an elimination–detoxification scheme with the known elimination rate constant. We concluded that quantification of trade-offs between subcellular partitioning and detoxification provides valuable insights into the ecotoxicology of aquatic organisms and enhances our understanding of the subcellular biology of trace metals.

Hierarchical regulation of mRNA partitioning between the cytoplasm and the endoplasmic reticulum of mammalian cells

The mRNA transcriptome is currently thought to be partitioned between the cytosol and endoplasmic reticulum (ER) compartments by binary selection; mRNAs encoding cytosolic/nucleoplasmic proteins are translated on free ribosomes, and mRNAs encoding topogenic signal-bearing proteins are translated on ER-bound ribosomes, with ER localization being conferred by the signal-recognition particle pathway. In subgenomic and genomic analyses of subcellular mRNA partitioning, we report an overlapping subcellular distribution of cytosolic/nucleoplasmic and topogenic signal-encoding mRNAs, with mRNAs of both cohorts displaying noncanonical subcellular partitioning patterns. Unexpectedly, the topogenic signal-encoding mRNA transcriptome was observed to partition in a hierarchical, cohort-specific manner. mRNAs encoding resident proteins of the endomembrane system were clustered at high ER-enrichment values, whereas mRNAs encoding secretory pathway cargo were broadly represented on free and ER-bound ribosomes. Two distinct modes of mRNA association with the ER were identified. mRNAs encoding endomembrane-resident proteins were bound via direct, ribosome-independent interactions, whereas mRNAs encoding secretory cargo displayed predominantly ribosome-dependent modes of ER association. These data indicate that mRNAs are partitioned between the cytosol and ER compartments via a hierarchical system of intrinsic and encoded topogenic signals and identify mRNA cohort-restricted modes of mRNA association with the ER.

Combining bioaccumulation and coping mechanism to enhance long-term site-specific risk assessment for zinc susceptibility of bivalves

The purpose of this study is to conduct a long-term site-specific risk assessment for zinc (Zn) susceptibility of bivalves, green mussel Perna viridis and hard clam Ruditapes philippinarum, based on published experimental data by linking the biologically-based damage assessment model with the subcellular partitioning concept. A comprehensive risk modeling framework was developed to predict susceptibility probability of two bivalve species exposed to waterborne Zn. The results indicated that P. viridis accumulates more Zn toxicity, whereas both toxic potency and the recovery rate of Zn are higher for R. philippinarum. We found that negative linear correlations exist in elimination–recovery and elimination–detoxification relationships, whereas a positive linear correlation was observed in recovery–detoxification relationships for bivalves exposed to waterborne Zn. Simulation results showed that the spatial differences of susceptibility primarily resulted from the variation of waterborne Zn concentration under field conditions. We found that R. philippinarum is more susceptible of Zn than P. viridis under the same exposure condition. Results also suggested that Zn posed no significant susceptibility risk to two bivalve species in Taiwan. We suggested that these two species can be used to biomonitor the water quality on Taiwan coastal areas.

Assessing abalone growth inhibition risk to cadmium and silver by linking toxicokinetics/toxicodynamics and subcellular partitioning

The purpose of this study was to link toxicokinetics/toxicodynamics and subcellular partitioning for assessing the susceptibility and the growth inhibition risks of abalone Haliotis diversicolor supertexta exposed to waterborne and foodborne cadmium (Cd) and silver (Ag). We reanalyzed published data on growth inhibition and subcellular partitioning associated with the present mechanistic model to explore the correlations among elimination (k (e)), detoxification (k (d)), and recovery (k (r)) rate constants and to assess the growth inhibition risk. We found a positive correlation among k (e), k (d), and k (r) in abalone exposed to Ag. We also employed a life-stage based probabilistic assessment model to estimate the growth inhibition risk of abalone to environmentally relevant Cd (5-995μgl(-1)) and Ag (0.05-9.95μgl(-1)) concentrations in Taiwan. The results showed that abalone had a minimum 20% probability of the growth inhibition risk exposed to Cd, whereas Ag exposure was not likely to pose the risk. The maximum biomasses were estimated to be 0.0039 and 0.0038, 61.61 and 43.87, and 98.88 and 62.97g for larvae, juveniles, and adults of abalone exposed to the same levels of Cd and Ag, respectively. Our study provides a useful tool to detect potential growth biomass of abalone populations subjected to Cd and Ag stresses and mechanistic implications for a long-term ecotoxicological risk assessment in realistic situations.