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Dendritic arborization is highly regulated and requires tight control of dendritic growth, branching, cytoskeletal dynamics, and ion channel expression to ensure proper function. Abnormal dendritic development can result in altered network connectivity, which has been linked to neurodevelopmental disorders, including autism spectrum disorders (ASDs). How neuronal growth control programs tune dendritic arborization to ensure function is still not fully understood. Using Drosophila dendritic arborization (da) neurons as a model, we identified the conserved Ste20-like kinase Tao as a negative regulator of dendritic arborization. We show that Tao kinase activity regulates cytoskeletal dynamics and sensory channel localization required for proper sensory function in both male and female flies. We further provide evidence for functional conservation of Tao kinase, showing that its ASD-linked human ortholog, Tao kinase 2 (Taok2), could replace Drosophila Tao and rescue dendritic branching, dynamic microtubule alterations, and behavioral defects. However, several ASD-linked Taok2 variants displayed impaired rescue activity, suggesting that Tao/Taok2 mutations can disrupt sensory neuron development and function. Consistently, we show that Tao kinase activity is required in developing and as well as adult stages for maintaining normal dendritic arborization and sensory function to regulate escape and social behavior. Our data suggest an important role for Tao kinase signaling in cytoskeletal organization to maintain proper dendritic arborization and sensory function, providing a strong link between developmental sensory aberrations and behavioral abnormalities relevant for Taok2-dependent ASDs.SIGNIFICANCE STATEMENT Autism spectrum disorders (ASDs) are linked to abnormal dendritic arbors. However, the mechanisms of how dendritic arbors develop to promote functional and proper behavior are unclear. We identified Drosophila Tao kinase, the ortholog of the ASD risk gene Taok2, a

The spine apparatus (SA) is an endoplasmic reticulum-related organelle that is present in a subset of dendritic spines in cortical and pyramidal neurons, and plays an important role in Ca2+ homeostasis and dendritic spine plasticity. The protein synaptopodin is essential for the formation of the SA and is widely used as a maker for this organelle. However, it is still unclear which factors contribute to its localization at selected synapses, and how it triggers local SA formation. In this study, we characterized development, localization and mobility of synaptopodin clusters in hippocampal primary neurons, as well as the molecular dynamics within these clusters. Interestingly, synaptopodin at the shaftassociated clusters is less dynamic than at spinous clusters. We identify the actin-based motor proteins myosin V (herein referring to both the myosin Va and Vb forms) and VI as novel interaction partners of synaptopodin, and demonstrate that myosin V is important for the formation and/or maintenance of the SA. We found no evidence of active microtubule-based transport of synaptopodin. Instead, new clusters emerge inside spines, which we interpret as the SA being assembled on-site.

Microtubules are hollow biopolymers of 25-nm diameter and are key constituents of the cytoskeleton. In neurons, microtubules are organized differently between axons and dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techniques can detect specific structures at an increased resolution, but the narrow spacing between neuronal microtubules poses challenges because most existing labelling strategies increase the effective microtubule diameter by 20-40 nm and will thereby blend neighbouring microtubules into one structure. Here we develop single-chain antibody fragments (nanobodies) against tubulin to achieve super-resolution imaging of microtubules with a decreased apparent diameter. To test the resolving power of these novel probes, we generate microtubule bundles with a known spacing of 50-70 nm and successfully resolve individual microtubules. Individual bundled microtubules can also be resolved in different mammalian cells, including hippocampal neurons, allowing novel insights into fundamental mechanisms of microtubule organization in cell- and neurobiology.

A high-performance magnetic resonance imaging T-2 contrast agent has been prepared via phase transfer of hydrophobic superparamagnetic iron oxide nanoparticles (SPIONs) to an aqueous solution by using Pluronic F127 copolymers. As seen in the figure, a hierarchical structure of the surface coating is formed and proven to be a crucial characteristic to enhance not only water dispersibility, but also the efficacy as a T-2 contrast agent., QC 20100721

The effect of the surface coating on the magnetic properties of superparamagnetic iron oxide nanoparticles (SPION) with 8 nm in size has been studied. Four different biocompatible coating layers are considered: poly L,L-lactic acid (PLLA), poly e-caprolactone (PCL), bovine serum albumin (BSA) and gold. The presence of coating layer on the surface of SPION is confirmed by FT-IR spectroscopy. Mössbauer spectroscopy and magnetic susceptibility measurements show that for uncoated SPION and Au@SPION the superparamagnetic fraction is retained. The formation of clusters in the case of BSA@SPION and chain-like structure for PCL@SPION and PLLA@SPION increase the inter-particle interactions resulting in hyperfine magnetic structure observed in the Mössbauer spectra at ambient temperature.

Nanoparticles, as building blocks, are important for the development of advanced, functional composite materials. Recent developments have shown that self-assembly of nanoparticles is a promising technique for the fabrication of complicate nanostructured materials. Self assembly of the nanoparticles into ordered structures on a substrate can be achieved through chemical treatment of the particle and/or substrate surface. The assembled nanoparticles can have a dramatic effect on the physical properties of the composite. A micronCP technique has been employed to form a SAM of bifunctional silane (APTMS) in the region of contact. The stamps for the micronCP are prepared by polymerization of polydimethylsiloxane (PDMS) on a flat surface. Glass substrates have been used for optical absorption measurements. Oxide or metallic particles have been assembled on the patterned surface after a surface treatment. The self-assembled layer was subsequently treated with bifunctional molecules and multilayers of the same material or composites have been thus obtained., This article is from the Mat. Res. Soc. Symposium Processing, v740 p217-222, 2003. This article is from ADA417952 Materials Research Society Symposium Proceedings Volume 740 Held in Boston, Massachusetts on December 2-6, 2002. Nanomaterials for Structural Applications

Superparamagnetic iron oxide nanoparticles (SPION) with suitable bio-compatible coatings have been used in biomedicine, particularly in magnetic resonance imaging (MRI), tissue engineering, and drug delivery applications. In this study, we describe the synthesis of SPION and its use for experimental in-vivo applications in MRI. SPION with a mean size of 6 nm have been prepared under inert atmosphere, in a polymeric starch matrix, by controlled chemical coprecipitation of magnetite phase from aqueous solutions containing suitable salts of Fe2+ and Fe3+. X-ray powder diffraction was used to confirm a pure magnetite phase for the SPION. The influence of oxidizing agents on the cleavage of the starch chains was investigated by changing the concentration of H2O2. An aqueous solution of H2O2/NaOH cleaves the glycosidic bonds and reduces the polymer chains to a critical average molecular weight. From the dynamic light scattering (DLS) size distribution, the bulk agglomeration size was decreased by approximately 50% of the bulk size when treated by H2O2. Freshly synthesized starch-coated SPION in buffered artificial cerebro-spinal fluid were injected into the brain parenchyma of anaesthetized rats for in-vivo monitoring. Analysis of T-2*-weighted images and T-2*-maps revealed formation of a. concentration gradient for the SPION at the injection site, indicating SPION dispersion in the living brain parenchyma from the center of the injection site toward the periphery. The starch-coated SPION show a biocompatibility and possibility of being transported in the extracellular space as well as being internalized in nerve cells., QC 20100525

Magnetic nanoparticles are becoming increasingly important for several biomedical applications. For example, superparamagnetic magnetite nanoparticles with suitable biocompatible coatings are useful in magnetic resonance imaging, tissue engineering, and drug delivery, etc. In this study we report the synthesis of magnetite nanoparticles and the further coating of these particles by several types of protective layers. Thermodynamic modeling of the chemical system has been adopted as a rational approach to establish routes to better synthesis conditions for pure phase magnetite. Quantitative analysis of different reaction equilibria involved in the precipitation of magnetite from aqueous solutions has been used to determine optimum synthesis conditions. Superparamagnetic magnetite nanoparticles (SPION) with diameters of 6 and 12 nm have been prepared by controlled chemical coprecipitation of magnetite phase from aqueous solutions containing suitable salts of Fe2+ and Fe3+ under inert atmosphere. Pure magnetite phase SPION could be observed from X-ray diffraction. Magnetic colloid suspensions containing particles with three different types of coatings (sodium oleate (NaOl), starch, and methoxypoly(ethylene glycol) (MPEG)) have been prepared by using different stabilization methods. SPION coatings were studied by determining the change of the surface charge by electrokinetic sonic amplitude (ESA) measurements, as a function of varying NaOl in the solution, where the amount of NaOl needed to form a stable suspension was determined. For stable suspension, the optimum concentration of sodium oleate (NaOl) chemisorbed at 2.5 g of SPION surface is 5.2 x 10(-7) M NaOl which shows maximum ESA value of 0.034 mPa(.)M/V. SPION coating by starch results in the formation of agglomerate. The agglomeration size of starch-coated SPION can be decreased by introducing H2O2 as an oxidizing agent; the resulting particle size is 42 nm as determined by dynamic light scattering (DLS). For th, QC 20100525