Bidirectional Transport Research Articles

Kinetic cooperativity resolves bidirectional clogging within the nuclear pore complex

As the main gatekeeper of the nucleocytoplasmic transport in eukaryotic cells, the nuclear pore complex (NPC) faces the daunting task of facilitating the bidirectional transport of a high volume of macromolecular cargoes while ensuring the selectivity, speed, and efficiency of this process. The competition between opposing nuclear import and export fluxes passing through the same channel is expected to pose a major challenge to transport efficiency. It has been suggested that phase separation-like radial segregation of import and export fluxes within the assembly of intrinsically disordered proteins that line the NPC pore could be a mechanism for ensuring efficient bidirectional transport. We examine the impact of radial segregation on the efficiency of bidirectional transport through the NPC using a coarse-grained computational model of the NPC. We find little evidence that radial segregation improves transport efficiency. By contrast, surprisingly, we find that NTR crowding may enhance rather than impair the efficiency of bidirectional transport although it decreases the available space in the pore. We identify mechanisms of this novel crowding-induced transport cooperativity through the self-regulation of cargo density and flux in the pore. These findings explain how the functional architecture of the NPC resolves the problem of efficient bidirectional transport, and provide inspiration for the alleviation of clogging in artificial selective nanopores.

Enhanced absorption of prenylated cinnamic acid derivatives from Brazilian green propolis by turmeric in humans and rats.

Prenylated cinnamic acid derivatives are the bioactive components of Brazilian green propolis (BGP). The effect of other botanical components on the pharmacokinetic profiles of these derivatives remains relatively unexplored. In the present study, we investigated the influence of several herbal extracts (turmeric, ginkgo leaf, coffee fruit, soybean, and gotu kola) on the plasma concentrations of cinnamic acid derivatives after BGP consumption. When the herbal extracts were co-administered with BGP in the clinical study, the area under the curve (AUC) values of artepillin C and drupanin, the major BGP components in plasma, were significantly increased by 1.7- and 1.5-fold, respectively, compared to those after BGP administration alone. Among the herbal extracts administered to rats, turmeric extract increased the AUC. Furthermore, a bidirectional transport assay suggested that artepillin C and drupanin are substrates of breast cancer resistance protein (BCRP), a drug elimination transporter. These results suggest that curcumin-containing turmeric extract may increase the plasma concentrations of artepillin C and drupanin via BCRP. Our findings enabled us to estimate the food-herb and herb-herb interactions invivo in foods and herbal medicines containing cinnamic acid derivatives and prenylated compounds.

Cooperation of Various Cytoskeletal Components Orchestrates Intercellular Spread of Mitochondria between B-Lymphoma Cells through Tunnelling Nanotubes.

Membrane nanotubes (NTs) are dynamic communication channels connecting spatially separated cells even over long distances and promoting the transport of different cellular cargos. NTs are also involved in the intercellular spread of different pathogens and the deterioration of some neurological disorders. Transport processes via NTs may be controlled by cytoskeletal elements. NTs are frequently observed membrane projections in numerous mammalian cell lines, including various immune cells, but their functional significance in the 'antibody factory' B cells is poorly elucidated. Here, we report that as active channels, NTs of B-lymphoma cells can mediate bidirectional mitochondrial transport, promoted by the cooperation of two different cytoskeletal motor proteins, kinesin along microtubules and myosin VI along actin, and bidirectional transport processes are also supported by the heterogeneous arrangement of the main cytoskeletal filament systems of the NTs. We revealed that despite NTs and axons being different cell extensions, the mitochondrial transport they mediate may exhibit significant similarities. Furthermore, we found that microtubules may improve the stability and lifespan of B-lymphoma-cell NTs, while F-actin strengthens NTs by providing a structural framework for them. Our results may contribute to a better understanding of the regulation of the major cells of humoral immune response to infections.

A Broadband Photodetector Based on Non-Layered MnS/WSe2 Type-I Heterojunctions with Ultrahigh Photoresponsivity and Fast Photoresponse.

The separation of photogenerated electron-hole pairs is crucial for the construction of high-performance and wide-band responsive photodetectors. The type-I heterojunction as a photodetector is seldomly studied due to its limited separation of the carriers and narrow optical response. In this work, we demonstrated that the high performance of type-I heterojunction as a broadband photodetector can be obtained by rational design of the band alignment and proper modulation from external electric field. The heterojunction device is fabricated by vertical stacking of non-layered MnS and WSe2 flakes. Its type-I band structure is confirmed by the first-principles calculations. The MnS/WSe2 heterojunction presents a wide optical detecting range spanning from 365 nm to 1550 nm. It exhibits the characteristics of bidirectional transportation, a current on/off ratio over 103, and an excellent photoresponsivity of 108 A W-1 in the visible range. Furthermore, the response time of the device is 19 ms (rise time) and 10 ms (fall time), which is much faster than that of its constituents MnS and WSe2. The facilitation of carrier accumulation caused by the interfacial band bending is thought to be critical to the photoresponse performance of the heterojunction. In addition, the device can operate in self-powered mode, indicating a photovoltaic effect.

Bidirectional ATP-driven transport of cobalamin by the mycobacterial ABC transporter BacA

BacA is a mycobacterial ATP-binding cassette (ABC) transporter involved in the translocation of water-soluble compounds across the lipid bilayer. Whole-cell-based assays have shown that BacA imports cobalamin as well as unrelated hydrophilic compounds such as the antibiotic bleomycin and the antimicrobial peptide Bac7 into the cytoplasm. Surprisingly, there are indications that BacA also mediates the export of different antibacterial compounds, which is difficult to reconcile with the notion that ABC transporters generally operate in a strictly unidirectional manner. Here we resolve this conundrum by developing a fluorescence-based transport assay to monitor the transport of cobalamin across liposomal membranes. We find that BacA transports cobalamin in both the import and export direction. This highly unusual bidirectionality suggests that BacA is mechanistically distinct from other ABC transporters and facilitates ATP-driven diffusion, a function that may be important for the evolvability of specific transporters, and may bring competitive advantages to microbial communities.

Insertases scramble lipids: Molecular simulations of MTCH2

Scramblases play a pivotal role in facilitating bidirectional lipid transport across cell membranes, thereby influencing lipid metabolism, membrane homeostasis, and cellular signaling. MTCH2, a mitochondrial outer membrane protein insertase, has a membrane-spanning hydrophilic groove resembling those that form the lipid transit pathway in known scramblases. Employing both coarse-grained and atomistic molecular dynamics simulations, we show that MTCH2 significantly reduces the free energy barrier for lipid movement along the groove and therefore can indeed function as a scramblase. Notably, the scrambling rate of MTCH2 in silico is similar to that of voltage-dependent anion channel (VDAC), a recently discovered scramblase of the outer mitochondrial membrane, suggesting a potential complementary physiological role for these mitochondrial proteins. Finally, our findings suggest that other insertases which possess a hydrophilic path across the membrane like MTCH2, can also function as scramblases.

Genome-Wide Identification and Expression Analysis of SWEET Gene Family in Strawberry

The Sugars Will Eventually be Exported Transporter (SWEET) is a class of bidirectional sugar transporter that is involved in critical physiological processes such as plant growth and development, and its response to biotic and abiotic stresses. Currently, there are few reports on the SWEET gene family in strawberry. In this study, we mined the SWEET gene family members in Fragaria × ananassa ‘Camarosa’ and carefully analyzed their molecular features and expression patterns. The results showed that 77 FanSWEET genes existed in the F. × ananassa ‘Camarosa’ genome, and the phylogenetic analysis classified them into four sub-groups. Analysis of gene structure, conserved structural domains, and conserved motifs showed that FanSWEETs were highly conserved during the evolutionary process. Expression profiling of the 11 FanSWEET genes revealed that three members were highly expressed in strawberry fruits, which were presumed to be involved in sugar transport during strawberry fruit ripening. In addition, based on the exogenous sugar-spraying treatment and quantitative real-time PCR analysis, we found that different members responded to different sugar treatments in different response patterns, and their functions in sugar transport need to be further explored. The present study provides a reference for further analysis of the functions of the SWEET gene in strawberry.

Chitosan and Anionic Solubility Enhancer Sulfobutylether-β-Cyclodextrin-Based Nanoparticles as Dexamethasone Ophthalmic Delivery System for Anti-Inflammatory Therapy.

Cataract surgery interventions are constantly increasing, particularly among adult and elderly patients. This type of surgery can lead to inflammatory states of the ocular anterior segment (AS), usually healed via postoperative treatment with dexamethasone (DEX)-containing eye drops. The application of eye drops is challenging due to the high number of daily administrations. In this study, mucoadhesive nanoparticles (NPs) were formulated to improve the residence time of DEX on the corneal mucosa, enhancing the drug's solubility and bioavailability. The NPs were generated using an ionotropic gelation technique, exploiting the interaction between the cationic group of chitosan (CS) and the anionic group of sulfobutylether-β-cyclodextrin (SBE-β-CD). The formation of the inclusion complex and its stoichiometry were studied through phase solubility studies, Job's plot method, and Bi-directional transport studies on MDCKII-MDR1. The obtained NPs showed good chemical and physical characteristics suitable for drug loading and subsequent testing on animal mucosa. The DEX-loaded CS/SBE-β-CD NPs exhibited a prolonged residence time on animal mucosa and demonstrated enhanced drug permeability through the corneal membrane, showing a sustained release profile. The developed NPs posed no irritation or toxicity concerns upon local administration, making them an optimal and innovative drug delivery system for inflammatory AS diseases treatment.

Plastic does not simply flow into the sea: River transport dynamics affected by tides and floating plants

Plastic pollution is ubiquitous in aquatic environments worldwide. Rivers connect terrestrial and marine ecosystems, playing a key role in the transport of land-based plastic waste towards the sea. Emerging research suggests that in estuaries and tidal rivers, tidal dynamics play a significant role in plastic transport and retention dynamics. To date, observations in these systems have been limited, and plastic transport dynamics during single tidal cycles remain poorly understood. Here, we investigated plastic transport, trapping, and re-mobilization of macroplastics (> 0.5 cm) in the Saigon River, focusing on short-term dynamics of individual tidal cycles. We used GPS trackers, released at different stages of the tidal cycle (ebb, flood, neap, spring). Plastic items demonstrated dynamic and intermittent transport behavior. Items spent almost half of the time (49%) temporarily stopped, mainly due to their entrapment in vegetation, infrastructure, or deposition on riverbanks. Items were almost always re-mobilized within 10 h (85%), leading to successive phases of stopping and transport. Tidal dynamics also resulted in bidirectional transport of plastic items, with median daily total transport distance within the 40 km study reach (8.9 km day−1) over four times larger than the median daily net distance (2.0 km day−1). The median retention time of plastic items within the reach was 21 days (mean = 202 days). In total, 81% of the retrieved items were trapped within water hyacinths, emphasizing the important role of floating vegetation on river plastic transport dynamics. With this paper, we aim to provide data-driven insights into macroplastic transport and retention dynamics in a tropical tidal river. These are crucial in the design of effective intervention and monitoring strategies, and estimating net plastic emission from rivers into the sea.

A DNA tensiometer for studying detachment kinetics of microtubule motors involved in bidirectional transport

A DNA tensiometer for studying detachment kinetics of microtubule motors involved in bidirectional transport

Robust behavior of kinesin and dynein motors in reconstituted bidirectional transport

Robust behavior of kinesin and dynein motors in reconstituted bidirectional transport

To be a transit link: Similarity in the structure of colonial system of integration and communication pores in autozooids and avicularia of Terminoflustra membranaceotruncata (Bryozoa: Cheilostomata).

Bryozoan colonies consist of zooids, which can differ in structure and function. Most heteromorphic zooids are unable to feed and autozooids supply them with nutrients. The structure of the tissues providing nutrient transfer is poorly investigated. Here, I present a detailed description of the colonial system of integration (CSI) and communication pores in autozooids and avicularia of the cheilosome bryozoan Terminoflustra membranaceotruncata. The CSI is the nutrient transport and distribution system in the colony. In both autozooids and avicularia it consists of a single cell type, that is, elongated cells, and has a variable branching pattern, except for the presence of a peripheral cord. The general similarity in the CSI structure in avicularia and autozooids is probably due to the interzooidal type of the avicularium. Interzooidal avicularia are likely to consume only a part of the nutrients delivered to them by the CSI, and they transit the rest of the nutrients further. The variability and irregularity of branching pattern of the CSI may be explained by the presence of single communication pores and their varying number. The structure of communication pores is similar regardless of their location (in the transverse or lateral wall) and the type of zooid in contact. Rosette complexes include a cincture cell, a few special cells, and a few limiting cells. Along each zooidal wall, there are communication pores with both unidirectional and bidirectional polarity of special cells. However, the total number of nucleus-containing lobes of special cells is approximately the same on each side of any zooidal wall. Supposing the polarity of special cells reflects the direction of nutrient transport, the pattern of special cells polarity is probably related to the need for bidirectional transport through each zooidal wall. The possibility for such transport is important in large perennial colonies with wide zones of autozooids undergoing polypide degeneration.

Tropomyosin 1-I/C coordinates kinesin-1 and dynein motors during oskar mRNA transport.

Dynein and kinesin motors mediate long-range intracellular transport, translocating towards microtubule minus and plus ends, respectively. Cargoes often undergo bidirectional transport by binding to both motors simultaneously. However, it is not known how motor activities are coordinated in such circumstances. In the Drosophila female germline, sequential activities of the dynein-dynactin-BicD-Egalitarian (DDBE) complex and of kinesin-1 deliver oskar messenger RNA from nurse cells to the oocyte, and within the oocyte to the posterior pole. We show through in vitro reconstitution that Tm1-I/C, a tropomyosin-1 isoform, links kinesin-1 in a strongly inhibited state to DDBE-associated oskar mRNA. Nuclear magnetic resonance spectroscopy, small-angle X-ray scattering and structural modeling indicate that Tm1-I/C suppresses kinesin-1 activity by stabilizing its autoinhibited conformation, thus preventing competition with dynein until kinesin-1 is activated in the oocyte. Our work reveals a new strategy for ensuring sequential activity of microtubule motors.

Quantitative comparison of nuclear transport inhibition by SARS coronavirus ORF6 reveals the importance of oligomerization

Open Reading Frame 6 (ORF6) proteins, which are unique to severe acute respiratory syndrome-related (SARS) coronavirus, inhibit the classical nuclear import pathway to antagonize host antiviral responses. Several alternative models were proposed to explain the inhibitory function of ORF6 [H. Xia et al., Cell Rep. 33, 108234 (2020); L. Miorin et al., Proc. Natl. Acad. Sci. U.S.A. 117, 28344-28354 (2020); and M. Frieman et al., J. Virol. 81, 9812-9824 (2007)]. To distinguish these models and build quantitative understanding of ORF6 function, we developed a method for scoring both ORF6 concentration and functional effect in single living cells. We combined quantification of untagged ORF6 expression level in single cells with optogenetics-based measurement of nuclear transport kinetics, using methods that could be adapted to measure concentration-dependent effects of any untagged protein. We found that SARS-CoV-2 ORF6 is ~15 times more potent than SARS-CoV-1 ORF6 in inhibiting nuclear import and export, due to differences in the C-terminal region that is required for the NUP98-RAE1 binding. The N-terminal region was required for transport inhibition. This region binds membranes but could be replaced by synthetic constructs which forced oligomerization in solution, suggesting its primary function is oligomerization. We propose that the hydrophobic N-terminal region drives oligomerization of ORF6 to multivalently cross-link the NUP98-RAE1 complexes at the nuclear pore complex, and this multivalent binding inhibits bidirectional transport.

Magnetically driven confined colloids: From enhanced diffusion to bidirectional transport

Inspired by previous experimental results, we use numerical simulations to investigate the collective dynamics of paramagnetic colloidal particles confined between two plates closer than twice the particle diameter and driven by an external precessing magnetic field. We show that, when the field is spatially isotropic there is no net particle current and the colloids display enhanced diffusive dynamics with an effective diffusion coefficient which raises up to 60 times that of the undriven case. In contrast, when the field is spatially anisotropic due to a small tilt angle δ, the particles organize into a robust bidirectional current, flowing along two parallel planes by periodically exchanging their positions. In this regime, we also analyze how the presence of small impurities which can be described as “magnetic holes” affect the particle current breaking the bidirectional flow. Our system provides a general method to transport magnetic colloids in a viscous fluid, without using any field gradient, but based on the fine balance between confinement and magnetic dipolar interactions.

Unidirectional and bidirectional photon transport blockade in driven atomic lattices of parity-time antisymmetry

We propose a scheme for realizing exact parity-time ( PT ) antisymmetry of complex susceptibility in a wide range of probe detuning by considering a one-dimensional lattice of cold atoms driven into the simplest three-level Λ configuration. This is attained by modulating the intensity of a standing-wave coupling field with a proper phase shift to counteract the product of a single-photon detuning and a two-photon detuning. Such a dynamically controlled PT -antisymmetric lattice supports the integration of a few nontrivial scattering behaviors including unidirectional light reflectionless, asymmetric perfect absorption, and directional signal quenching. These behaviors, depending in particular on atomic densities and lattice lengths, facilitate the on-demand realization of unidirectional or bidirectional photon transport blockade.

Using Live-Cell Imaging to Measure the Effects of Pathological Proteins on Axonal Transport in Primary Hippocampal Neurons.

Bidirectional transport of cargos along the axon is critical for maintaining functional synapses, neural connectivity, and healthy neurons. Axonal transport is disrupted in multiple neurodegenerative diseases, and projection neurons are particularly vulnerable because of the need to transport cellular materials over long distances and sustain substantial axonal mass. Pathological modifications of several disease-related proteins negatively affect transport, including tau, amyloid-β, α-synuclein, superoxide dismutase, and huntingtin, providing a potential common mechanism by which pathological proteins exert toxicity in disease. Methods to study these toxic mechanisms are necessary to understand neurodegenerative disorders and identify potential therapeutic interventions. Here, cultured primary rodent hippocampal neurons are co-transfected with multiple plasmids to study the effects of pathological proteins on fast axonal transport using live-cell confocal imaging of fluorescently-tagged cargo proteins. We begin with the harvest, dissociation, and culturing of primary hippocampal neurons from rodents. Then, we co-transfect the neurons with plasmid DNA constructs to express fluorescent-tagged cargo protein and wild-type or mutant tau (used as an exemplar of pathological proteins). Axons are identified in live cells using an antibody that binds an extracellulardomain of neurofascin, an axon initial segment protein, and an axonal region of interest is imaged to measure fluorescent cargo transport. Using KymoAnalyzer, a freely available ImageJ macro, we extensively characterize the velocity, pause frequency, and directional cargo density of axonal transport, all of which may be affected by the presence of pathological proteins. Through this method, we identify a phenotype of increased cargo pause frequency associated with the expression of pathological tau protein. Additionally, gene-silencing shRNA constructs can be added to the transfection mix to test the role of other proteins in mediating transport disruption. This protocol is easily adaptable for use with other neurodegenerative disease-related proteins and is a reproducible method to study the mechanisms of how those proteins disrupt axonal transport.

Kapβ2 Inhibits Perioperative Neurocognitive Disorders in Rats with Mild Cognitive Impairment by Reversing the Nuclear-Cytoplasmic Mislocalization of hnRNPA2/B1.

Harmful stimuli trigger mutations lead to uncontrolled accumulation of hnRNPA2/B1 in the cytoplasm, exacerbating neuronal damage. Kapβ2 mediates the bidirectional transport of most substances between the cytoplasm and the nucleus. Kapβ2 guides hnRNPA2/B1 back into the nucleus and restores its function, alleviating related protein toxicity. Here, we aim to explore the involvement of Kapβ2 in neurodegeneration in rats with MCI following sevoflurane anesthesia and surgery. Firstly, novel object recognition test and Barnes maze were conducted to assess behavioral performances, and we found Kapβ2 positively regulated the recovery of memory and cognitive function. In vivo electrophysiological experiments revealed that the hippocampal theta rhythm energy distribution was disrupted, coherence was reduced, and long-term potentiation was attenuated in MCI rats. LTP was greatly improved with positive modulation of Kapβ2. Next, functional MRI and BOLD imaging will be employed to examine the AFLL and FC values of dynamic connectivity between the cortex and hippocampus of the brain. The findings show that regulating Kapβ2 in the hippocampus region enhances functional activity and connections between brain regions in MCI rats. WB results showed that increasing Kapβ2 expression improved the expression and recovery of cognitive-related proteins in the hippocampus of MCI rats. Finally, WB and immunofluorescence were used to examine the changes in hnRNPA2/B1 expression in the nucleus and cytoplasm after overexpression of Kapβ2, and it was found that nucleocytoplasmic mis location was alleviated. Overall, these data show that Kapβ2 reverses the nucleoplasmic misalignment of hnRNPA2/B1, which slows neurodegeneration towards dementia in MCI after sevoflurane anesthesia and surgery. Our findings may lead to new approaches for perioperative neuroprotection of MCI patients.

Bidirectional tick transport by migratory birds of the African-Western Palearctic flyway over Turkish Thrace: observation of the current situation and future projection.

This study was carried out at a vital stopover site of migrating birds in the Turkish Thrace, European part of Turkey, on the Mediterranean/Black Sea Flyway. Ticks were collected from the birds captured in the four migration periods, i.e., autumn 2020, spring 2021, autumn 2021, and spring 2022, and identified morphologically. Throughout the study, 10,651 birds from 77 species were examined, and 671 belonging to 34 species were found infested. The infestation prevalence in total birds and the mean number of ticks per infested bird were 6.3% and 3.8 (range: 1-142), respectively. A total of 2573 ticks were collected with the following species distribution and numbers: Ixodes spp. 70 larvae, I. frontalis 1829 larvae, 337 nymphs, and 30 adults, I. acuminatus 16 nymphs and 42 adults, I. ricinus 39 larvae, 141 nymphs, and one adult, Hyalomma spp. seven larvae and 60 nymphs, and Haemaphysalis sp. one larva. Prevalence, intensity, and species distribution of the ticks in birds varied depending on the month, season, year, and species-specific migration phenology of the birds. The results show that precise determination of the tick-borne risk associated with migratory birds for a particular region necessarily requires long-term and comprehensive studies and indicates that anthropogenic climate change and habitat degradation can significantly differentiate the risk by influencing the migration phenology in birds and by making new regions suitable for the establishment of different ticks.

Transplacental transport of per- and polyfluoroalkyl substances (PFAS): Mechanism exploration via BeWo cell monolayer model

Per- and polyfluoroalkyl substances (PFAS) have received global concern on adverse effects on pregnancy outcomes. Although human studies have reported fetal exposure to PFAS, the underlying mechanisms driving transplacental transfer of PFAS have not been sufficiently understood. The present study aimed to investigate chemical-specific transplacental transfer of PFAS and potential mechanisms based on a BeWo cell monolayer model. The findings of concentration- and time-dependent transport, asymmetry in bidirectional transport, molecular docking and transporter inhibition experiments indicate that passive diffusion and membrane transporter-involved active transport could collectively determine transplacental transport of PFAS. Membrane transporters could play important roles in chemical-specific transport. The inhibition of OAT transporter resulted in promotion of trans-monolayer transport for most PFAS, while an opposite trend was observed when P-gp, BCRP and MRP transporters were prohibited. By contrast, inhibition of OCT resulted in inhibitory effects on the transport of some PFAS (i.e., PFHxA, PFHpA, PFOA, and PFNA), and promotive effects on the other substances (i.e., PFUdA, PFHpS, PFOS, 6:2 Cl-PFESA and PFOSA). Therefore, simultaneous involvement of diverse membrane transporters in utero could result in complicated influence on transplacental transport. Our work constitutes a solid ground for further exploration of the effects of gestational PFAS exposure on birth outcomes.