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The unique morphology of neurons consists of a long axon and a highly variable arbour of dendritic processes, which assort neuronal cells into the main classes. The dendritic tree serves as the main domain for receiving synaptic input. Therefore, to maintain the structure and to be able to plastically change according to the incoming stimuli, molecules and organelles need to be readily available. This is achieved mainly via bi-directional transport of cargo along the microtubule lattices. Analysis of dendritic transport is lagging behind the investigation of axonal transport. Moreover, addressing transport mechanisms in tissue environment is very challenging and, therefore, rare. We employed high-speed volumetric lattice light-sheet microscopy and single particle tracking of truncated KIF1A motor protein lacking the cargo-binding domain. We focused our analysis on dendritic processes of CA1 pyramidal neurons in cultured hippocampal tissue. Analysis of individual trajectories revealed detailed information about stalling and high variability in movement and speed, and biased directionality of KIF1A. Furthermore, we could also observe KIF1A shortly entering into dendritic spines. We provide a workflow to analyse variations in the speed and direction of motor protein movement in dendrites that are either intrinsic properties of the motor domain or depend on the structure and modification of the microtubule trails. [ABSTRACT FROM AUTHOR]

Background: Hereditary spastic paraplegia (HSP) is a heterogeneous group of inherited neurodegenerative disorders characterized by slowly progressive lower limbs spasticity and weakness. HSP type 30 (SPG30) is a HSP subtype caused by mutations in the kinesin family member 1A gene (KIF1A) and could be either autosomal dominantly or recessively inherited. The aim of this study was to investigate the clinical and genetic features of KIF1A mutations in a Taiwanese HSP cohort.Methods: Mutational analysis of KIF1A was performed in 242 unrelated Taiwanese patients of Han Chinese ethnicity with clinically suspected HSP using targeted resequencing panel covering the entire coding regions of KIF1A. Clinical, electrophysiological and neuroimaging features of the HSP patients carrying a KIF1A mutation were characterized.Results: Three different KIF1A mutations were identified in three patients with autosomal dominantly inherited HSP. Among them, KIF1A p.E19K was a novel mutation. The patient harboring KIF1A p.G321D presented with pure HSP, while the individuals carrying KIF1A p.E19K or p.R316Q manifested complex HSP with additional axonal sensorimotor polyneuropathy. The patients carrying KIF1A p.R316Q also had thoracic cord atrophy, thin corpus callosum and white matter hyperintensity.Conclusion: SPG30 accounts for 1.2% (3/242) of patients in the Taiwanese HSP cohort, suggesting that it is an uncommon HSP subtype in Taiwan. This study delineates the clinical and genetic features of SPG30 in Taiwan and provides useful information for the diagnosis and management of SPG30, especially in patients of Han Chinese descent. [ABSTRACT FROM AUTHOR]

Neuroblastoma (NB) comprises about 8–10% of pediatric cancers, and microRNA (miR)-338 downregulation has been implicated in NB. However, the underlying molecular mechanism remains largely unclear. The main goal of this study is to probe the regulatory role of miR-338 and the upstream and downstream biomolecules involved in NB. The differentially expressed miRNAs were screened by analyzing the NB gene expression microarray GSE121513 from the GEO database, and the differences in expression of the screened miRNAs were verified in clinically collected NB tissues versus dorsal root ganglions. Subsequently, the relationship between the miR-338 expression and NB cell growth was validated in vitro and in vivo , and the upstream and downstream regulatory mechanisms of miR-338 were further analyzed by bioinformatics. Functional rescue experiments were used to verify their effects on NB cell growth. miR-338 expressed poorly in NB tissues, and overexpression of miR-338 significantly inhibited NB cell growth in vitro and in vivo. The prediction results showed that miR-338 could target KIF1A, and miR-338 expression was negatively correlated with the expression of KIF1A. We further found that miR-338 was transcriptionally regulated by the transcription factor KLF5. Overexpression of KLF5 or KIF1A significantly attenuated the inhibitory effect of miR-338 mimic on NB cell growth. Finally, miR-338 blocked the Hedgehog signaling pathway by inhibiting the expression of KIF1A. Overexpression of KLF5 reduced expression of miR-338, which in turn increased the expression of KIF1A and activated the Hedgehog signaling pathway, leading to the progression of NB. [ABSTRACT FROM AUTHOR]

Summary Tight regulation of neuronal transport allows for cargo binding and release at specific cellular locations. The mechanisms by which motor proteins are loaded on vesicles and how cargoes are captured at appropriate sites remain unclear. To better understand how KIF1A-driven dense core vesicle (DCV) transport is regulated, we identified the KIF1A interactome and focused on three binding partners, the calcium binding protein calmodulin (CaM) and two synaptic scaffolding proteins: liprin-α and TANC2. We showed that calcium, acting via CaM, enhances KIF1A binding to DCVs and increases vesicle motility. In contrast, liprin-α and TANC2 are not part of the KIF1A-cargo complex but capture DCVs at dendritic spines. Furthermore, we found that specific TANC2 mutations—reported in patients with different neuropsychiatric disorders—abolish the interaction with KIF1A. We propose a model in which Ca 2+ /CaM regulates cargo binding and liprin-α and TANC2 recruit KIF1A-transported vesicles. [ABSTRACT FROM AUTHOR]

• We identified a inframe KIF1A mutation in a family with autosomal dominant HSP. • This mutation is located in the non-motor domain of the KIF1A gene. • The proband shows a milder phenotype than other SPG30 phenotypes. SPG30 is a newly categorized type of HSP caused by variants in the kinesin family member 1A gene (KIF1A). Advances in next-generation sequencing have resulted in a limited number of studies describing the clinical, electrophysiological, and radiological features of HSP, with variable manifestations. Most known pathogenic KIF1A variants affect the motor domain, although some rare pathogenic variants have been identified that affect the non-motor domain. Here, we report a Korean family with a rare homozygous autosomal-recessive form of SPG30. A 59-year-old man and his father presented with an uncomplicated, mild SPG30 phenotype, characterized by a progressive, spastic gait. Familial co-segregation analysis revealed a pathogenic c.2751_2753delGGA KIF1A variant that affects the non-motor domain. Our case broadens the genetic and clinical variability of SPG30, warranting similar studies to consolidate the pathogenicity of SPG30. [ABSTRACT FROM AUTHOR]

Abstract: Dense-core vesicles (DCVs) are responsible for transporting, processing, and secreting neuropeptide cargos that mediate a wide range of biological processes, including neuronal development, survival, and learning and memory. DCVs are synthesized in the cell body and are transported by kinesin motor proteins along microtubules to pre- and postsynaptic release sites. Due to the dependence on kinesin-based transport, we sought to determine if the kinesin-3 family member, KIF1A, transports DCVs in primary cultured hippocampal neurons, as has been described for invertebrate neurons. Two-color, live-cell imaging showed that the DCV markers, chromogranin A-RFP and BDNF-RFP, move together with KIF1A-GFP in both the anterograde and retrograde directions. To demonstrate a functional role for KIF1A in DCV transport, motor protein expression in neurons was reduced using RNA interference (shRNA). Fluorescently tagged DCV markers showed a significant reduction in organelle flux in cells expressing shRNA against KIF1A. The transport of cargo driven by motors other than KIF1A, including mitochondria and the transferrin receptor, was unaffected in KIF1A shRNA expressing cells. Taken together, these data support a primary role for KIF1A in the anterograde transport of DCVs in mammalian neurons, and also provide evidence that KIF1A remains associated with DCVs during retrograde DCV transport. [Copyright &y& Elsevier]

• Dynamic microtubules enter dendritic spines in an activity-dependent manner, where they contribute to structural plasticity. • The microtubule cytoskeleton acts as a scaffold at inhibitory postsynaptic sites, where it controls the "influx" and "efflux" of receptors during synaptic plasticity. • Presynaptic microtubules play multiple roles in bouton organization, local synaptic vesicle trafficking and mitochondrial arrangement in the terminal. • Synaptic microtubule dysfunction may underlie neurological disease. In neurons, microtubules (MTs) provide routes for transport throughout the cell and structural support for dendrites and axons. Both stable and dynamic MTs are necessary for normal neuronal functions. Research in the last two decades has demonstrated that MTs play additional roles in synaptic structure and function in both pre- and postsynaptic elements. Here, we review current knowledge of the functions that MTs perform in excitatory and inhibitory synapses, as well as in the neuromuscular junction and other specialized synapses, and discuss the implications that this knowledge may have in neurological disease [ABSTRACT FROM AUTHOR]

Tissue homeostasis depends on precise yet plastic regulation of stem cell daughter fates. During growth, Drosophila intestinal stem cells (ISCs) adjust fates by switching from asymmetric to symmetric lineages to scale the size of the ISC population. Using a combination of long-term live imaging, lineage tracing, and genetic perturbations, we demonstrate that this switch is executed through the control of mitotic spindle orientation by Jun-N-terminal kinase (JNK) signaling. JNK interacts with the WD40-repeat protein Wdr62 at the spindle and transcriptionally represses the kinesin Kif1a to promote planar spindle orientation. In stress conditions, this function becomes deleterious, resulting in overabundance of symmetric fates and contributing to the loss of tissue homeostasis in the aging animal. Restoring normal ISC spindle orientation by perturbing the JNK/Wdr62/Kif1a axis is sufficient to improve intestinal physiology and extend lifespan. Our findings reveal a critical role for the dynamic control of SC spindle orientation in epithelial maintenance. • Spindles reorient from oblique to planar during growth, driving symmetric outcomes • Stress- and aging-mediated activation of JNK promotes planar spindles • JNK interacts with Wdr62 and represses Kif1a expression to promote planar spindles • Restoring oblique spindles in old flies extends lifespan Hu and Jasper demonstrate that spindle orientation regulates stem cell fate in Drosophila intestines. JNK activity promotes reorientation from oblique to planar spindles, increasing symmetric outcomes. This switch is required for growth but becomes chronic during stress and age. Restoring oblique spindles in old animals improves tissue physiology, extending lifespan. [ABSTRACT FROM AUTHOR]