Retrograde Direction Research Articles

Quantitatively Dissecting Triple Roles of Dynactin in Dynein-Driven Transport of Influenza Virus by Quantum Dot-Based Single-Virus Tracking.

After entering host cells by endocytosis, influenza A virus (IAV) is transported along microfilaments and then transported by dynein along microtubules (MTs) to the perinuclear region for genome release. Understanding the mechanisms of dynein-driven transport is significant for a comprehensive understanding of IAV infection. In this work, the roles of dynactin in dynein-driven transport of IAV were quantitatively dissected in situ using quantum dot-based single-virus tracking. It was revealed that dynactin was essential for dynein to transport IAV toward the nucleus. After virus entry, virus-carrying vesicles bound to dynein and dynactin before being delivered to MTs. The attachment of dynein to the vesicles was dependent on dynactin and its subunits, p150Glued and Arp1. Once viruses reached MTs, dynactin-assisted dynein initiates retrograde transport of IAV. Importantly, the retrograde transport of viruses could be initiated at both plus ends (32%) and other regions on MTs (68%). Subsequently, dynactin accompanied and assisted dynein to persistently transport the virus along MTs in the retrograde direction. This study revealed the dynactin-dependent dynein-driven transport process of IAV, enhancing our understanding of IAV infection and providing important insights into the cell's endocytic transport mechanism.

Duodenal ileus caused by a cage magnet in a Fleckvieh cow.

The present report aims to describe the case of a duodenal obstruction ileus in a dairy cow that was caused by a cage magnet. The 4.7-year-old German Fleckvieh cow was hospitalized because of symptoms of intestinal obstruction such as anorexia, noticeable drop in milk yield, reduced defecation, dehydration and positive percussion and swinging auscultation in a circumscribed area cranial of the right flank over the last 2 ribs. Six months as well as 3 days prior to hospitalization the cow had already been treated for signs of hardware disease, which included administration of a cage magnet.After the initial clinical diagnostic procedure on hospital admission, a diagnostic laparotomy in the right paralumbar fossa was performed to identify the cause of the ileus. The cranial part of the duodenum was markedly dilated, and a solid foreign body was found obstructing the intestine immediately aboral to the duodenal sigmoid flexure. This was identified as a cage magnet, which was massaged in retrograde direction into the pyloric antrum and removed via abomasotomy. The cow recovered from surgical intervention and was discharged from the hospital 6 days later.The present report describes an unusual complication of cage magnet administration, which is a standard veterinary procedure and generally considered a safe treatment option in cows with clinical signs of acute traumatic reticuloperitonitis.

Axonal autophagic vesicle transport in the rat optic nerve in vivo under normal conditions and during acute axonal degeneration

Neurons pose a particular challenge to degradative processes like autophagy due to their long and thin processes. Autophagic vesicles (AVs) are formed at the tip of the axon and transported back to the soma. This transport is essential since the final degradation of the vesicular content occurs only close to or in the soma. Here, we established an in vivo live-imaging model in the rat optic nerve using viral vector mediated LC3-labeling and two-photon-microscopy to analyze axonal transport of AVs. Under basal conditions in vivo, 50% of the AVs are moving with a majority of 85% being transported in the retrograde direction. Transport velocity is higher in the retrograde than in the anterograde direction. A crush lesion of the optic nerve results in a rapid breakdown of retrograde axonal transport while the anterograde transport stays intact over several hours. Close to the lesion site, the formation of AVs is upregulated within the first 6 h after crush, but the clearance of AVs and the levels of lysosomal markers in the adjacent axon are reduced. Expression of p150Glued, an adaptor protein of dynein, is significantly reduced after crush lesion. In vitro, fusion and colocalization of the lysosomal marker cathepsin D with AVs are reduced after axotomy. Taken together, we present here the first in vivo analysis of axonal AV transport in the mammalian CNS using live-imaging. We find that axotomy leads to severe defects of retrograde motility and a decreased clearance of AVs via the lysosomal system.

Plasma dynamics of individual HiPIMS pulses: imaging study using high-frame-rate camera

A high-frame-rate camera with microsecond-level time resolution was used to make systematic investigations of plasma self-organization and spoke dynamics during individual HiPIMS pulses. The plasma was imaged for a range of argon pressures (0.25–2 Pa) and peak discharge currents (10–400 A) using an Al target. The experiments revealed that plasma evolves through three characteristic stages as the discharge current increases. In stage I, which is present from the current onset and up to ∼25 A, spokes are azimuthally long and rotate in the −Ez × B direction. The spoke behavior is similar to the one observed in DCMS discharges. The number of spokes depends on pressure and the current growth rate. At the lowest pressure (0.25 Pa) a single spoke is present in discharge, while at higher pressures (1–2 Pa) two spokes are most often observed. The spoke velocity depends on the number of spokes, current growth rate and pressure. A single spoke rotates with velocities in the 4–15 km s−1 range, while two spokes rotate in the 1–9 km s−1 range depending on the pressure and growth rate. Following stage I, the plasma undergoes a complex reorganization that is characterized by aperiodic spoke patterns and irregular dynamics. In stage II spokes are less localized, they merge, split and propagate either in the retrograde or prograde direction. After chaotic plasma reorganization, more ordered spoke patterns begin to form. Spokes in stage III are azimuthally shorter, typically exhibit a triangular shape and rotate in the Ez × B direction. In general, the spoke dynamics is less complicated and is only influenced by the pressure. Spokes rotate faster at higher pressures than at lower ones; velocities range from 9 km s−1 at 0.25 Pa to 6 km s−1 at 2 Pa. The spoke velocity in stage III is largely unaffected by the discharge current or number of spokes. Stage III can be further divided into sub-stages, which are characterized by different current growth rates, spoke sizes and shapes. In general, the spoke evolution is highly reproducible for pulses with similar discharge current waveforms.

Efficient axonal transport of endolysosomes relies on the balanced ratio of microtubule tyrosination and detyrosination.

In neurons, the microtubule (MT) cytoskeleton forms the basis for long-distance protein transport from the cell body into and out of dendrites and axons. To maintain neuronal polarity, the axon initial segment (AIS) serves as a physical barrier, separating the axon from the somatodendritic compartment and acting as a filter for axonal cargo. Selective trafficking is further instructed by axonal enrichment of MT post-translational modifications, which affect MT dynamics and the activity of motor proteins. Here, we compared two knockout mouse lines lacking the respective enzymes for MT tyrosination and detyrosination, and found that both knockouts led to a shortening of the AIS. Neurons from both lines also showed an increased immobile fraction of endolysosomes present in the axon, whereas mobile organelles displayed shortened run distances in the retrograde direction. Overall, our results highlight the importance of maintaining the balance of tyrosinated and detyrosinated MTs for proper AIS length and axonal transport processes.

A Linear Model for Inertial Modes in a Differentially Rotating Sun

Inertial wave modes in the Sun are of interest owing to their potential to reveal new insight into the solar interior. These predominantly retrograde-propagating modes in the solar subsurface appear to deviate from the thin-shell Rossby–Haurwitz model at high azimuthal orders. We present new measurements of sectoral inertial modes at m > 15 where the modes appear to become progressively less retrograde compared to the canonical Rossby–Haurwitz dispersion relation in a corotating frame. We use a spectral eigenvalue solver to compute the spectrum of solar inertial modes in the presence of differential rotation. Focussing specifically on equatorial Rossby modes, we find that the numerically obtained mode frequencies lie along distinct ridges, one of which lies strikingly close to the observed mode frequencies in the Sun. We also find that the n = 0 ridge is deflected strongly in the retrograde direction. This suggests that the solar measurements may not correspond to the fundamental n = 0 Rossby–Haurwitz solutions as was initially suspected, but to those for a higher n. The numerically obtained eigenfunctions also appear to sit deep within the convection zone—unlike those for the n = 0 modes—which differs substantially from solar measurements and complicates inference.

Swedish Alzheimer’s disease variant perturbs activity of retrograde molecular motors and causes widespread derangement of axonal transport pathways

Experimental studies in flies, mice, and humans suggest a significant role of impaired axonal transport in the pathogenesis of Alzheimer's disease (AD). The mechanisms underlying these impairments in axonal transport, however, remain poorly understood. Here we report that the Swedish familial AD mutation causes a standstill of the amyloid precursor protein (APP) in the axons at the expense of its reduced anterograde transport. The standstill reflects the perturbed directionality of the axonal transport of APP, which spends significantly more time traveling in the retrograde direction. This ineffective movement is accompanied by an enhanced association of dynactin-1 with APP, which suggests that reduced anterograde transport of APP is the result of enhanced activation of the retrograde molecular motor dynein by dynactin-1. The impact of the Swedish mutation on axonal transport is not limited to the APP vesicles since it also reverses the directionality of a subset of early endosomes, which become enlarged and aberrantly accumulate in distal locations. In addition, it also reduces the trafficking of lysosomes due to their less effective retrograde movement. Altogether, our experiments suggest a pivotal involvement of retrograde molecular motors and transport in the mechanisms underlying impaired axonal transport in AD and reveal significantly more widespread derangement of axonal transport pathways in the pathogenesis of AD.

Multispacecraft Observations of a Widespread Solar Energetic Particle Event on 2022 February 15–16

On 2022 February 15–16, multiple spacecraft measured one of the most intense solar energetic particle (SEP) events observed so far in Solar Cycle 25. This study provides an overview of interesting observations made by multiple spacecraft during this event. Parker Solar Probe (PSP) and BepiColombo were close to each other at 0.34–0.37 au (a radial separation of ∼0.03 au) as they were impacted by the flank of the associated coronal mass ejection (CME). At about 100° in the retrograde direction and 1.5 au away from the Sun, the radiation detector on board the Curiosity surface rover observed the largest ground-level enhancement on Mars since surface measurements began. At intermediate distances (0.7–1.0 au), the presence of stream interaction regions (SIRs) during the SEP arrival time provides additional complexities regarding the analysis of the distinct contributions of CME-driven versus SIR-driven events in observations by spacecraft such as Solar Orbiter and STEREO-A, and by near-Earth spacecraft like ACE, SOHO, and WIND. The proximity of PSP and BepiColombo also enables us to directly compare their measurements and perform cross-calibration for the energetic particle instruments on board the two spacecraft. Our analysis indicates that energetic proton measurements from BepiColombo and PSP are in reasonable agreement with each other to within a factor of ∼1.35. Finally, this study introduces the various ongoing efforts that will collectively improve our understanding of this impactful, widespread SEP event.

Dynein and dynactin move long-range but are delivered separately to the axon tip.

Axonal transport is essential for neuronal survival. This is driven by microtubule motors including dynein, which transports cargo from the axon tip back to the cell body. This function requires its cofactor dynactin and regulators LIS1 and NDEL1. Due to difficulties imaging dynein at a single-molecule level, it is unclear how this motor and its regulators coordinate transport along the length of the axon. Here, we use a neuron-inducible human stem cell line (NGN2-OPTi-OX) to endogenously tag dynein components and visualize them at a near-single molecule regime. In the retrograde direction, we find that dynein and dynactin can move the entire length of the axon (>500 µm). Furthermore, LIS1 and NDEL1 also undergo long-distance movement, despite being mainly implicated with the initiation of dynein transport. Intriguingly, in the anterograde direction, dynein/LIS1 moves faster than dynactin/NDEL1, consistent with transport on different cargos. Therefore, neurons ensure efficient transport by holding dynein/dynactin on cargos over long distances but keeping them separate until required.

Numerical study of non-toroidal inertial modes withl=m+ 1 radial vorticity in the Sun’s convection zone

Various types of inertial modes have been observed and identified on the Sun, including the equatorial Rossby modes, critical-latitude modes, and high-latitude modes. Recent observations have further reported the detection of equatorially antisymmetric radial vorticity modes that propagate in a retrograde direction about three times faster than those of the equatorial Rossby modes, when seen in the corotating frame with the Sun. Here, we study the properties of these equatorially antisymmetric vorticity modes using a realistic linear model of the Sun’s convection zone. We find that they are essentially non-toroidal, involving a substantial radial flow at the equator. Thus, the background density stratification plays a critical role in determining their dispersion relation. The solar differential rotation is also found to have a significant impact by introducing the viscous critical layers and confining the modes near the base of the convection zone. Furthermore, we find that their propagation frequencies are strikingly sensitive to the background superadiabaticity,δ, because the buoyancy force acts as an additional restoring force for these non-toroidal modes. The observed frequencies are compatible with the linear model only when the bulk of the convection zone is weakly subadiabatic (−5 × 10−7 ≲ δ ≲ −2.5 × 10−7). Our result is consistent with but tighter than the constraint independently derived in a previous study (δ < 2 × 10−7), employing the high-latitude inertial mode. It is implied that, below the strongly superadiabatic near-surface layer, the bulk of the Sun’s convection zone might be much closer to adiabatic than typically assumed or it may even be weakly subadiabatic.

CD8α-CI-M6PR Particle Motility Assay to Study the Retrograde Motion of CI-M6PR Receptors in Cultured Living Cells.

The cation-independent mannose 6-phosphate receptors (CI-M6PR) bind newly synthesized mannose 6-phosphate (Man-6-P)-tagged enzymes in the Golgi and transport them to late endosomes/lysosomes, providing them with degradative functions. Following the cargo delivery, empty receptors are recycled via early/recycling endosomes back to the trans-Golgi network (TGN) retrogradely in a dynein-dependent motion. One of the most widely used methods for studying the retrograde trafficking of CI-M6PR involves employing the CD8α-CI-M6PR chimera. This chimera, comprising a CD8 ectodomain fused with the cytoplasmic tail of the CI-M6PR receptor, allows for labeling at the plasma membrane, followed by trafficking only in a retrograde direction. Previous studies utilizing the CD8α-CI-M6PR chimera have focused mainly on colocalization studies with various endocytic markers under steady-state conditions. This protocol extends the application of the CD8α-CI-M6PR chimera to live cell imaging, followed by a quantitative analysis of its motion towards the Golgi. Additionally, we present an approach to quantify parameters such as speed and track lengths associated with the motility of CD8α-CI-M6PR endosomes using the Fiji plugin TrackMate. Key features • This assay is adapted from the methodology by Prof. Matthew Seaman for studying the retrograde trafficking of CI-M6PR by expressing CD8α-CI-M6PR chimera in HeLa cells. • The experiments include live-cell imaging of surface-labeled CD8α-CI-M6PR molecules, followed by a chase in cells. • Allows the monitoring of real-time motion of CD8α-CI-M6PR endosomes and facilitates calculation of kinetic parameters associated with endosome trajectories, e.g., speed and distance (run lengths).

Dyneins

Dyneins are a family of motor proteins that carry out motility and force generation functions towards the minus end of microtubule filaments. Cytoplasmic dynein (dynein-1) is responsible for transporting intracellular cargos in the retrograde direction in the cytoplasm, anchoring several organelles to the microtubule network, driving nuclear migration in developing neurons, and orienting the mitotic spindle in dividing cells. All other dyneins are localized to cilia. Similar to dynein-1, dynein-2 walks along microtubules and drives intraflagellar transport in the retrograde direction. Other ciliary dyneins are positioned between adjacent microtubule doublets of the axoneme and power ciliary beating by sliding microtubules relative to each other. In this primer, we first highlight the structure, mechanism, and regulation of dynein-1, which is the best-characterized member of the dynein motor family, and then describe the unique features and cellular roles of other dyneins. We also discuss accessory proteins that regulate the activation and motility of dynein motors in different cellular contexts.

Spotlight on Trans-Synaptic Degeneration in the Visual Pathway in Multiple Sclerosis.

A putative mechanism of neurodegeneration in multiple sclerosis (MS) is trans-synaptic degeneration (TSD), whereby injury to a neuron leads to degeneration of synaptically connected neurons. The visual system is commonly involved in MS and provides an ideal model to study TSD given its well-defined structure. TSD may occur in an anterograde direction (optic neuropathy causing degeneration in the posterior visual pathway including the optic radiations and occipital gray matter) and/or retrograde direction (posterior visual pathway lesions causing retinal degeneration). In the current review, we discuss evidence supporting the presence of anterograde and retrograde TSD in the visual system in MS.

Retrograde discs around one component of a binary are unstable to tilting

ABSTRACT With hydrodynamic simulations, we show that a coplanar disc around one component of a binary can be unstable to global tilting when the disc orbits in a retrograde direction relative to the binary. The disc experiences the largest inclination growth relative to the binary orbit in the outermost radii of the disc, closest to the companion. This tilt instability also occurs for test particles. A retrograde disc is much larger than a prograde disc since it is not tidally truncated and instead spreads outwards to the orbit of the companion. The coplanar retrograde disc remains circular while a coplanar prograde disc can become eccentric. We suggest that the inclination instability is due to a disc resonance caused by the interaction of the tilt with the tidal field of the binary. This model is applicable to Be/X-ray binaries in which the Be star disc may be retrograde relative to the binary orbit if there was a sufficiently strong kick from the supernova that formed the neutron star companion. The accretion on to the neutron star and the resulting X-ray outbursts are weaker in the retrograde case compared to the prograde case.

Scalarized Kerr-Newman black holes

In this paper, we construct scalarized rotating black holes within the framework of Einstein-Maxwell-scalar models. These models incorporate non-minimal couplings that can induce tachyonic instabilities, leading to the spontaneous scalarization of Kerr-Newman (KN) black holes. By exploring the domain of existence, we observe that the presence of scalarized KN black holes is suppressed by the black hole spin, with a maximum spin threshold beyond which scalarized solutions cease to exist. Intriguingly, we find that in specific parameter regimes, scalarized KN black holes can exhibit the presence of two unstable and one stable light rings on the equatorial plane, manifesting in both prograde and retrograde directions.

Dextran Amine-Conjugated Neural Tracing in Mosquitoes.

To understand the circuitry of the brain, it is often advantageous to visualize the processes of a single neuron or population of neurons. Identifying sites where a neuron, or neurons, originates and where it projects can allow a researcher to begin to map the circuitry underlying various processes, including sensory-guided behaviors. Furthermore, neural tracing allows one to map locations where processes terminate onto regions of the brain that may have known functions and sometimes to identify candidate upstream or downstream connections, based on proximity. Many methods of neural tracing are available; here, we focus on loading fluorescent dyes into a neuron (fluorescent dye filling). Different options for dyes exist to label neurites. Among the most versatile and easy to use are dextran amine-conjugated dyes. They fill neurons bidirectionally, not discriminating between anterograde or retrograde loading direction. Dye filling must be done in unfixed tissue, as the dye needs to move through the neurons; however, dextran amine conjugates are aldehyde-fixable and once cells have been fully loaded with dye the tissue can be fixed and subjected to immunostaining. Coupling neural tracing with immunofluorescence is a useful way to determine specific brain or ventral nerve cord (VNC) regions where a neuron projects. This protocol describes methods for loading dextran amine conjugated dyes into a sensory tissue in the mosquito to visualize sites of sensory neuron innervation in the central nervous system, as well as efferent projections to these structures. This protocol is described for Aedes aegypti, for which it was optimized, but it also works across a variety of insects.

Experimental and numerical study of flow structure in a model of distal anastomosis of femoral artery

Physical and numerical simulation of the steady blood flow was carried out focusing on the distal end-to-side anastomosis of a graft to the femoral artery. The angle between the graft and the artery was 60°. Two Reynolds numbers were considered that corresponded to the physiological range when estimated using the average (Re = 240) and maximum (Re = 1640) blood flow through the human femoral artery over a single cardiac cycle. The experiment included flow visualization and measurements of instantaneous vector fields of velocity using an optical SIV (Smoke Image Velocimetry) technique. Direct numerical simulation (DNS) was employed for numerical study. The distribution of blood flows through the branches of the main artery was 80 % in the antegrade and 20 % in the retrograde direction. Main patterns in the distribution of velocity and its root-mean-square fluctuations in both branches of the main artery were revealed. Flow separation regions were observed in the branching area. Additionally, secondary flows (Prandtl’s vortices of the first kind) were revealed. The flow in the branching area was still laminar Re = 240, while the signs of transition to turbulence were documented within separation regions at Re = 1640. Distributions of streamwise and circumferential components of skin friction vector were estimated. In some regions of flow, these components were shown to be comparable in size. The location of regions was revealed in which the friction magnitude in the considered flow regimes was lower than the one in an unimpaired artery (developed laminar flow in a circular pipe) at appropriate Reynolds numbers.

Hippocampal damage causes retrograde amnesia for objects' visual, but not odour, properties in male rats.

Damage to the hippocampus produces profound retrograde amnesia, but odour and object discrimination memories can be spared in the retrograde direction. Prior lesion studies testing retrograde amnesia for object/odour discriminations are problematic due to sparing of large parts of the hippocampus, which may support memory recall, and/or the presence of uncontrolled, distinctive odours that may support object discrimination. To address these issues, we used a simple object discrimination test to assess memory in male rats. Two visually distinct objects, paired with distinct odour cues, were presented. One object was associated with a reward. Following training, neurotoxic hippocampal lesions were made using N-methyl-D-aspartate (NMDA). The rats were then tested on the preoperatively learned object discrimination problem, with and without the availability of odour or visual cues during testing. The rats were also postoperatively trained on a new object discrimination problem. Lesion sizes ranged from 67% to 97% of the hippocampus (average of 87%). On the preoperatively learned discrimination problem, the rats with hippocampal lesions showed preserved object discrimination memory when tested in the dark (i.e., without visual cues) but not when the explicit odour cues were removed from the objects. Hippocampal lesions increased the number of trials required to reach criterion but did not prevent rats from solving the postoperatively learned discrimination problem. Our results support the idea that long-term memories for odours, unlike recall of visual properties of objects, do not depend on the hippocampus in rats, consistent with previous observations that hippocampal damage does not cause retrograde amnesia for odour memories.

WDR31 displays functional redundancy with GTPase-activating proteins (GAPs) ELMOD and RP2 in regulating IFT complex and recruiting the BBSome to cilium.

The correct intraflagellar transport (IFT) assembly at the ciliary base and the IFT turnaround at the ciliary tip are key for the IFT to perform its function, but we still have poor understanding about how these processes are regulated. Here, we identify WDR31 as a new ciliary protein, and analysis from zebrafish and Caenorhabditis elegans reveals the role of WDR31 in regulating the cilia morphology. We find that loss of WDR-31 together with RP-2 and ELMD-1 (the sole ortholog ELMOD1-3) results in ciliary accumulations of IFT Complex B components and KIF17 kinesin, with fewer IFT/BBSome particles traveling along cilia in both anterograde and retrograde directions, suggesting that the IFT/BBSome entry into the cilia and exit from the cilia are impacted. Furthermore, anterograde IFT in the middle segment travels at increased speed in wdr-31;rpi-2;elmd-1 Remarkably, a non-ciliary protein leaks into the cilia of wdr-31;rpi-2;elmd-1, possibly because of IFT defects. This work reveals WDR31-RP-2-ELMD-1 as IFT and BBSome trafficking regulators.

Brain-wide N2cG compensation permits glycoprotein-deleted rabies virus to trace neural circuits across multiple synapses

Rabies-viruses-based retrograde tracers can spread across multiple synapses in a retrograde direction in the nervous system of rodents and primates, making them powerful tools for determining the structure and function of the complicated neural circuits of the brain. However, they have some limitations, such as posing high risks to human health and the inability to retrograde trans-synaptic label inputs from genetically-defined starter neurons. Here, we established a new retrograde trans-multi-synaptic tracing method through brain-wide rabies virus glycoprotein (RVG) compensation, followed by glycoprotein-deleted rabies virus (RV-[Formula: see text]G) infection in specific brain regions. Furthermore, in combination with the avian tumor virus receptor A (TVA) controlled by a cell-type-specific promoter, we found that EnvA-pseudotyped RV-[Formula: see text]G can mediate efficient retrograde trans-multi-synaptic transduction from cell-type-specific starter neurons. This study provides new alternative methods for neuroscience researchers to analyze the input neural networks of rodents and nonhuman primates.