Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy

Background: Cytoplasmic mislocalization and aggregation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of the amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD) disease spectrum, causing both nuclear loss-of-function and cytoplasmic toxic gain-of-function phenotypes. While TDP-43 proteinopathy has been associated with defects in nucleocytoplasmic transport, this process is still poorly understood. Here we study the role of karyopherin-β1 (KPNB1) and other nuclear import receptors in regulating TDP-43 pathology. Methods: We used immunostaining, immunoprecipitation, biochemical and toxicity assays in cell lines, primary neuron and organotypic mouse brain slice cultures, to determine the impact of KPNB1 on the solubility, localization, and toxicity of pathological TDP-43 constructs. Postmortem patient brain and spinal cord tissue was stained to assess KPNB1 colocalization with TDP-43 inclusions. Turbidity assays were employed to study the dissolution and prevention of aggregation of recombinant TDP-43 fibrils in vitro. Fly models of TDP-43 proteinopathy were used to determine the effect of KPNB1 on their neurodegenerative phenotype in vivo. Results: We discovered that several members of the nuclear import receptor protein family can reduce the formation of pathological TDP-43 aggregates. Using KPNB1 as a model, we found that its activity depends on the prion-like C-terminal region of TDP-43, which mediates the co-aggregation with phenylalanine and glycine-rich nucleoporins (FG-Nups) such as Nup62. KPNB1 is recruited into these co-aggregates where it acts as a molecular chaperone that reverses aberrant phase transition of Nup62 and TDP-43. These findings are supported by the discovery that Nup62 and KPNB1 are also sequestered into pathological TDP-43 aggregates in ALS/FTD postmortem CNS tissue, and by the identification of the fly ortholog of KPNB1 as a strong protective modifier in Drosophila models of TDP-43 proteinopathy. Our results show that KPNB1 can rescue all hallmarks of TDP-43 pathology, by restoring its solubility and nuclear localization, and reducing neurodegeneration in cellular and animal models of ALS/FTD. Conclusion: Our findings suggest a novel NLS-independent mechanism where, analogous to its canonical role in dissolving the diffusion barrier formed by FG-Nups in the nuclear pore, KPNB1 is recruited into TDP-43/FG-Nup co-aggregates present in TDP-43 proteinopathies and therapeutically reverses their deleterious phase transition and mislocalization, mitigating neurodegeneration. Graphical Abstract: [Figure not available: see fulltext.]

The JCMT BISTRO Survey: An 850/450 μ m Polarization Study of NGC 2071IR in Orion B

Abstract: We present the results of simultaneous 450 μm and 850 μm polarization observations toward the massive star-forming region NGC 2071IR, a target of the BISTRO (B-fields in STar-forming Region Observations) Survey, using the POL-2 polarimeter and SCUBA-2 camera mounted on the James Clerk Maxwell Telescope. We find a pinched magnetic field morphology in the central dense core region, which could be due to a rotating toroidal disklike structure and a bipolar outflow originating from the central young stellar object IRS 3. Using the modified Davis–Chandrasekhar–Fermi method, we obtain a plane-of-sky magnetic field strength of 563 ± 421 μG in the central ∼0.12 pc region from 850 μm polarization data. The corresponding magnetic energy density of 2.04 × 10−8 erg cm−3 is comparable to the turbulent and gravitational energy densities in the region. We find that the magnetic field direction is very well aligned with the whole of the IRS 3 bipolar outflow structure. We find that the median value of polarization fractions is 3.0% at 450 μm in the central 3′ region, which is larger than the median value of 1.2% at 850 μm. The trend could be due to the better alignment of warmer dust in the strong radiation environment. We also find that polarization fractions decrease with intensity at both wavelengths, with slopes, determined by fitting a Rician noise model of 0.59 ± 0.03 at 450 μm and 0.36 ± 0.04 at 850 μm, respectively. We think that the shallow slope at 850 μm is due to grain alignment at the center being assisted by strong radiation from the central young stellar objects

Progressive Changes in MOR1 in the Striatum of the Heterozygous Q175 Knock-in Mouse Model of Huntington's Disease

Huntington’s disease (HD) is a progressive autosomal dominant neurodegenerative disease that produces pronounced neurodegeneration of neostriatal neurons as well as cortical neurons in HD patients. Despite the fact that HD is caused by a dominant mutation in a single gene, the HTT gene, HD patients display great phenotypic variability related to both motor and mood symptoms. In the early stages of HD, there is a massive preferential loss of medium spiny neurons and asymmetry in the degeneration of the striatal compartments, particularly in the striosomal system (Thu et al., 2010). However, the role of striosomes in relation to HD mouse models still has yet to be elucidated. This study investigates whether the striosome system in the striatum is disrupted in a mouse model of Huntington’s disease (HD), the Q175 HD mouse model. In the human brain, the most striking neuropathology is the significant loss of the medium spiny neurons (MSNs) in the striatum and cortical pyramidal neurons, relative to the neuronal loss in other regions. Post-mortem tissue analysis in both human and animal models of HD, show that the striosome and matrix striatal compartments degenerate in an asymmetrical manner. Could the striosomes be a direct player to the dysfunctions that occur in HD? Furthermore, the striosome neurons are rich in mu-opioid receptors, which are involved in regulating dopamine functions in the brain. Could the disruption of the striosome system explain the changes occurring in the dopamine system in HD? I explored these questions by examining the immunohistochemical and immunofluorescence staining patterns and distributions of mu-opioid receptor 1 (MOR1) in transverse brain sections of the Q175 HD model mice. The initial results suggest an augmentation of the MOR1, a possible upregulation of the receptor, indicating a dysregulation of the striosome-enriched receptor protein. Post-mortem analysis of HD brains has shown that individuals with preferential degeneration of striosomes were more likely to have experienced predominant mood symptoms. These imbalances in the striosome system could contribute to the range of emotional and cognitive problems experienced by HD patients

In Vivo Detection of Extracellular Adenosine Triphosphate in a Mouse Model of Traumatic Brain Injury

none9Traumatic brain injury (TBI) is traditionally characterized by primary and secondary injury phases, both contributing to pathological and morphological changes. The mechanisms of damage and chronic consequences of TBI remain to be fully elucidated, but synaptic homeostasis disturbances and impaired energy metabolism are proposed to be a major contributor. It has been proposed that an increase of extracellular (eATP) adenosine triphosphate (ATP) in the area immediately surrounding impact may play a pivotal role in this sequence of events. After tissue injury, rupture of cell membranes allows release of intracellular ATP into the extracellular space, triggering a cascade of toxic events and inflammation. ATP is a ubiquitous messenger; however, simple and reliable techniques to measure its concentration have proven elusive. Here, we integrate a sensitive bioluminescent eATP sensor known as pmeLUC, with a controlled cortical impact mouse model to monitor eATP changes in a living animal after injury. Using the pmeLUC probe, a rapid increase of eATP is observed proximal to the point of impact within minutes of the injury. This event is significantly attenuated when animals are pretreated with an ATP hydrolyzing agent (apyrase) before surgery, confirming the contribution of eATP. This new eATP reporter could be useful for understanding the role of eATP in the pathogenesis in TBI and may identify a window of opportunity for therapeutic intervention.noneFaroqi, Ayman H; Lim, Melina J; Kee, Emma C; Lee, Jannifer H; Burgess, Jeremy D; Chen, Ridong; Di Virgilio, Francesco; Delenclos, Marion; McLean, Pamela JFaroqi, Ayman H; Lim, Melina J; Kee, Emma C; Lee, Jannifer H; Burgess, Jeremy D; Chen, Ridong; Di Virgilio, Francesco; Delenclos, Marion; Mclean, Pamela

Radial Glial Lineage Progression and Differential Intermediate Progenitor Amplification Underlie Striatal Compartments and Circuit Organization

Summary The circuitry of the striatum is characterized by two organizational plans: the division into striosome and matrix compartments, thought to mediate evaluation and action, and the direct and indirect pathways, thought to promote or suppress behavior. The developmental origins of these organizations and their developmental relationships are unknown, leaving a conceptual gap in understanding the cortico-basal ganglia system. Through genetic fate mapping, we demonstrate that striosome-matrix compartmentalization arises from a lineage program embedded in lateral ganglionic eminence radial glial progenitors mediating neurogenesis through two distinct types of intermediate progenitors (IPs). The early phase of this program produces striosomal spiny projection neurons (SPNs) through fate-restricted apical IPs (aIPSs) with limited capacity; the late phase produces matrix SPNs through fate-restricted basal IPs (bIPMs) with expanded capacity. Notably, direct and indirect pathway SPNs arise within both aIPS and bIPM pools, suggesting that striosome-matrix architecture is the fundamental organizational plan of basal ganglia circuitry