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1.
Soft Matter ; 18(10): 2060-2071, 2022 Mar 09.
Article En | MEDLINE | ID: mdl-35199113

Plants, animals, and humans use camouflage to blend in with their surroundings. The camouflage is achieved with different combinations of colors, patterns, and morphologies. In stealth applications, the simplest camouflage uses textiles colored similarly to the environment to create an illusion. However, often, visible light range camouflage is not enough since the multispectral detection technologies of today are readily utilized for identification. Foams can be created with a straightforward fabricating process, and lightweight material exhibits good thermal insulation properties, providing stealth in the infrared light region. Herein, we produce cellulosic wet foams from surfactant and bleached pulp or cellulose nanofibrils. The visible light camouflage is created with green microalgae, Chlorella vulgaris, and brown kraft lignin, which also stabilized the foams. The thermal and spectral camouflage performance of foams was influenced by the cellulose content as well as the stability and water content of foams. Overall, these results give insight into how stability impacts the thermal and spectral properties of wet foams and provide a solid base for further material development to improve camouflage performance. While there is plenty of data on dry foams, the functional behavior of wet foams is currently not well known. Our method, using plant-based components can be exploited in a variety of other applications where simplicity and scalability are important.


Chlorella vulgaris , Microalgae , Cellulose , Lignin
2.
Sci Rep ; 11(1): 12646, 2021 06 16.
Article En | MEDLINE | ID: mdl-34135443

A polydimethylsiloxane (PDMS)/Cu superhydrophobic composite material is fabricated by wet etching, electroless plating, and polymer casting. The surface topography of the material emerges from hierarchical micro/nanoscale structures of etched aluminum, which are rigorously copied by plated copper. The resulting material is superhydrophobic (contact angle > 170°, sliding angle < 7° with 7 µL droplets), electrically conductive, elastic and wear resistant. The mechanical durability of both the superhydrophobicity and the metallic conductivity are the key advantages of this material. The material is robust against mechanical abrasion (1000 cycles): the contact angles were only marginally lowered, the sliding angles remained below 10°, and the material retained its superhydrophobicity. The resistivity varied from 0.7 × 10-5 Ωm (virgin) to 5 × 10-5 Ωm (1000 abrasion cycles) and 30 × 10-5 Ωm (3000 abrasion cycles). The material also underwent 10,000 cycles of stretching and bending, which led to only minor changes in superhydrophobicity and the resistivity remained below 90 × 10-5 Ωm.

3.
Micromachines (Basel) ; 11(11)2020 Oct 30.
Article En | MEDLINE | ID: mdl-33143339

A hypoxic (low oxygen level) microenvironment and nitric oxide paracrine signaling play important roles in the control of both biological and pathological cell responses. In this study, we present a microfluidic chip architecture for nitric oxide delivery under a hypoxic microenvironment in human embryonic kidney cells (HEK-293). The chip utilizes two separate, but interdigitated microfluidic channels. The hypoxic microenvironment was created by sodium sulfite as the oxygen scavenger in one of the channels. The nitric oxide microenvironment was created by sodium nitroprusside as the light-activated nitric oxide donor in the other channel. The solutions are separated from the cell culture by a 30 µm thick gas-permeable, but liquid-impermeable polydimethylsiloxane membrane. We show that the architecture is preliminarily feasible to define the gaseous microenvironment of a cell culture in the 100 µm and 1 mm length scales.

4.
ACS Appl Mater Interfaces ; 12(44): 49957-49962, 2020 Nov 04.
Article En | MEDLINE | ID: mdl-33084313

This work presents a superhydrophobic antireflective (AR) coating on glass. The coating consists of a grass-like alumina layer capped with plasma-deposited fluoropolymer. The grass-like alumina is formed by hot water treatment of atomic layer-deposited alumina on glass, and the fluoropolymer is plasma-deposited from CHF3. Excellent broadband AR performance is observed in the visible spectrum with an average transmission of 94.9% for single-sided coated glass, which is close to the maximum 95.3% possible for this glass. Extremely desirable contact angles are obtained with 5-7 min-long fluoropolymer treatments on grass-like alumina with 173° advancing and 160° receding contact angles. This type of multifunctional coating can be beneficial in a multitude of applications like self-cleaning AR coating for solar panels, windows in high-rise buildings, sensors, and aerospace applications as well as just utilizing the excellent water repellent behavior in applications where only superhydrophobicity is required.

5.
Sci Rep ; 10(1): 17661, 2020 10 19.
Article En | MEDLINE | ID: mdl-33077786

Different types of carbon materials are biocompatible with neural cells and can promote maturation. The mechanism of this effect is not clear. Here we have tested the capacity of a carbon material composed of amorphous sp3 carbon backbone, embedded with a percolating network of sp2 carbon domains to sustain neuronal cultures. We found that cortical neurons survive and develop faster on this novel carbon material. After 3 days in culture, there is a precocious increase in the frequency of neuronal activity and in the expression of maturation marker KCC2 on carbon films as compared to a commonly used glass surface. Accelerated development is accompanied by a dramatic increase in neuronal dendrite arborization. The mechanism for the precocious maturation involves the activation of intracellular calcium oscillations by the carbon material already after 1 day in culture. Carbon-induced oscillations are independent of network activity and reflect intrinsic spontaneous activation of developing neurons. Thus, these results reveal a novel mechanism for carbon material-induced neuronal survival and maturation.


Calcium/metabolism , Carbon , Cell Differentiation , Neurons/physiology , Dendrites/physiology , Humans , Nerve Net , Neurons/metabolism
6.
Macromol Rapid Commun ; 41(2): e1900522, 2020 Jan.
Article En | MEDLINE | ID: mdl-31778287

Tunable and responsive surfaces offer routes to multiple functionalities ranging from superhydrophobic surfaces to controlled adhesion. Inspired by cilia structure in the respiratory pathway, magnetically responsive periodic arrays of flexible and magnetic thiol-ene micropillars are fabricated. Omnidirectional collective bending of the pillar array in magnetic field is shown. Local non-contact actuation of a single pillar is achieved using an electromagnetic needle to probe the responsiveness and the elastic properties of the pillars by comparing the effect of thiol-ene crosslinking density to pillar bending. The suitable thiol-ene components for flexible and stiff magnetic micropillars and the workable range of thiol-to-allyl ratio are identified. The wettability of the magnetic pillars can be tailored by chemical and topography modification of the pillar surface. Low-surface-energy self-assembled monolayers are grafted by UV-assisted surface activation, which is also used for surface topography modification by covalent bonding of micro- and nanoparticles to the pillar surface. The modified thiol-ene micopillars are resistant to capillarity-driven collapse and they exhibit low contact angle hysteresis, allowing water droplet motion driven by repeated bending and recovery of the magnetic pillars in an external magnetic field. Transport of polyethylene microspheres is also demonstrated.


Allyl Compounds/chemistry , Click Chemistry , Magnetite Nanoparticles/chemistry , Sulfhydryl Compounds/chemistry , Elasticity , Electromagnetic Fields , Hydrophobic and Hydrophilic Interactions , Particle Size , Propylene Glycols/chemistry , Sulfides/chemical synthesis , Sulfides/chemistry , Surface Properties , Triazines/chemistry , Water/chemistry , Wettability
7.
Glob Chall ; 3(2): 1800079, 2019 Feb.
Article En | MEDLINE | ID: mdl-31565363

Point-of-care devices that are inexpensive, disposable, and environmentally friendly are becoming increasingly predominant in the field of biosensing and biodiagnostics. Here, microfluidics is a suitable option to endow portability and minimal reagent and material consumption. Nanocellulose is introduced to manufacture microfluidic channels and as a storage and immobilization compartment of glucose oxidase. Improved enzymatic activity retention is demonstrated in a simple and disposable point-of-care diagnostic unit that is able to detect glucose from fluid matrices at 0.1 × 10-3 m concentration and in less than 10 min. It is concluded that the patterning and fluidic technologies that are possible with nanocellulose enable easily scalable multianalyte designs.

8.
Micromachines (Basel) ; 10(8)2019 Jul 31.
Article En | MEDLINE | ID: mdl-31370267

Pattern formation of pyrolyzed carbon (PyC) and tetrahedral amorphous carbon (ta-C) thin films were investigated at micro- and nanoscale. Micro- and nanopillars were fabricated from both materials, and their biocompatibility was studied with cell viability tests. Carbon materials are known to be very challenging to pattern. Here we demonstrate two approaches to create biocompatible carbon features. The microtopographies were 2 µ m or 20 µ m pillars (1:1 aspect ratio) with three different pillar layouts (square-grid, hexa-grid, or random-grid orientation). The nanoscale topography consisted of random nanopillars fabricated by maskless anisotropic etching. The PyC structures were fabricated with photolithography and embossing techniques in SU-8 photopolymer which was pyrolyzed in an inert atmosphere. The ta-C is a thin film coating, and the structures for it were fabricated on silicon substrates. Despite different fabrication methods, both materials were formed into comparable micro- and nanostructures. Mouse neural stem cells were cultured on the samples (without any coatings) and their viability was evaluated with colorimetric viability assay. All samples expressed good biocompatibility, but the topography has only a minor effect on viability. Two µ m pillars in ta-C shows increased cell count and aggregation compared to planar ta-C reference sample. The presented materials and fabrication techniques are well suited for applications that require carbon chemistry and benefit from large surface area and topography, such as electrophysiological and -chemical sensors for in vivo and in vitro measurements.

9.
Sci Rep ; 9(1): 5189, 2019 Mar 26.
Article En | MEDLINE | ID: mdl-30914658

Studies in optics and acoustics have employed metamaterial lenses to achieve sub-wavelength localization, e.g. a recently introduced concept called 'acoustojet' which in simulations localizes acoustic energy to a spot smaller than λ/2. However previous experimental results on the acoustojet have barely reached λ/2-wide localization. Here we show, by simulations and experiments, that a sub-λ/2 wide localization can be achieved by translating the concept of a photonic jet into the acoustic realm. We performed nano- to macroscale molecular dynamics (MD) and finite element method (FEM) simulations as well as macroscale experiments. We demonstrated that by choosing a suitable size cylindrical lens, and by selecting the speed-of-sound ratio between the lens material(s) and the surrounding medium, an acoustic jet ('acoustic sheet') is formed with a full width at half maximum (FWHM) less than λ/2. The results show, that the acoustojet approach can be experimentally realized with easy-to-manufacture acoustic lenses at the macroscale. MD simulations demonstrate that the concept can be extended to coherent phonons at nanoscale. Finally, our FEM simulations identify some micrometer size structures that could be realized in practice. Our results may contribute to starting a new era of super resolution acoustic imaging: We foresee that jet generating constructs can be readily manufactured, since suitable material combinations can be found from nanoscale to macroscale. Tight focusing of mechanical energy is highly desirable in e.g. electronics, materials science, medicine, biosciences, and energy harvesting.

10.
Micromachines (Basel) ; 9(12)2018 Dec 19.
Article En | MEDLINE | ID: mdl-30572576

The low fabrication cost of SU-8-based devices has opened the fields of point-of-care devices (POC), µTAS and Lab-on-Chip technologies, which call for cheap and disposable devices. Often this translates to free-standing, suspended devices and a reusable carrier wafer. This necessitates a sacrificial layer to release the devices from the substrates. Both inorganic (metals and oxides) and organic materials (polymers) have been used as sacrificial materials, but they fall short for fabrication and releasing multilayer SU-8 devices. We propose photoresist AZ 15nXT (MicroChemicals GmbH, Ulm, Germany) to be used as a sacrificial layer. AZ 15nXT is stable during SU-8 processing, making it suitable for fabricating free-standing multilayer devices. We show two methods for cross-linking AZ 15nXT for stable sacrificial layers and three routes for sacrificial release of the multilayer SU-8 devices. We demonstrate the capability of our release processes by fabrication of a three-layer free-standing microfluidic electrospray ionization (ESI) chip and a free-standing multilayer device with electrodes in a microchannel.

11.
ACS Sustain Chem Eng ; 6(11): 14631-14640, 2018 Nov 05.
Article En | MEDLINE | ID: mdl-30416892

In the current study, platinum-present as a negligible component (below 1 ppb, the detection limit of the HR-ICP-MS at the dilutions used) in real industrial hydrometallurgical process solutions-was recovered by an electrodeposition-redox replacement (EDRR) method on pyrolyzed carbon (PyC) electrode, a method not earlier applied to metal recovery. The recovery parameters of the EDRR process were initially investigated using a synthetic nickel electrolyte solution ([Ni] = 60 g/L, [Ag] = 10 ppm, [Pt] = 20 ppm, [H2SO4] = 10 g/L), and the results demonstrated an extraordinary increase of 3 × 105 in the [Pt]/[Ni] on the electrode surface cf. synthetic solution. EDRR recovery of platinum on PyC was also tested with two real industrial process solutions that contained a complex multimetal solution matrix: Ni as the major component (>140 g/L) and very low contents of Pt, Pd, and Ag (i.e., <1 ppb, 117 and 4 ppb, respectively). The selectivity of Pt recovery by EDRR on the PyC electrode was found to be significant-nanoparticles deposited on the electrode surface comprised on average of 90 wt % platinum and a [Pt]/[Ni] enrichment ratio of 1011 compared to the industrial hydrometallurgical solution. Furthermore, other precious metallic elements like Pd and Ag could also be enriched on the PyC electrode surface using the same methodology. This paper demonstrates a remarkable advancement in the recovery of trace amounts of platinum from real industrial solutions that are not currently considered as a source of Pt metal.

12.
Commun Biol ; 1: 97, 2018.
Article En | MEDLINE | ID: mdl-30271977

The engineering of microbial systems increasingly strives to achieve a co-existence and co-functioning of different populations. By creating interactions, one can utilize combinations of cells where each population has a specialized function, such as regulation or sharing of metabolic burden. Here we describe a microfluidic system that enables long-term and independent growth of fixed and distinctly separate microbial populations, while allowing communication through a thin nano-cellulose filter. Using quorum-sensing signaling, we can couple the populations and show that this leads to a rapid and stable connection over long periods of time. We continue to show that this control over communication can be utilized to drive nonlinear responses. The coupling of separate populations, standardized interaction, and context-independent function lay the foundation for the construction of increasingly complex community-wide dynamic genetic regulatory mechanisms.

13.
Acta Biomater ; 73: 167-179, 2018 06.
Article En | MEDLINE | ID: mdl-29649636

Physiological oxygen levels within the tissue microenvironment are usually lower than 14%, in stem cell niches these levels can be as low as 0-1%. In cell cultures, such low oxygen levels are usually mimicked by altering the global culture environment either by O2 removal (vacuum or oxygen absorption) or by N2 supplementation for O2 replacement. To generate a targeted cellular hypoxic microenvironment under ambient atmospheric conditions, we characterised the ability of the dissolved oxygen-depleting sodium sulfite to generate an in-liquid oxygen sink. We utilised a microfluidic design to place the cultured cells in the vertical oxygen gradient and to physically separate the cells from the liquid. We demonstrate generation of a chemical in-liquid oxygen sink that modifies the surrounding O2 concentrations. O2 level control in the sink-generated hypoxia gradient is achievable by varying the thickness of the polydimethylsiloxane membrane. We show that intracellular hypoxia and hypoxia response element-dependent signalling is instigated in cells exposed to the microfluidic in-liquid O2 sink-generated hypoxia gradient. Moreover, we show that microfluidic flow controls site-specific microenvironmental kinetics of the chemical O2 sink reaction, which enables generation of intermittent hypoxia/re-oxygenation cycles. The microfluidic O2 sink chip targets hypoxia to the cell culture microenvironment exposed to the microfluidic channel architecture solely by depleting O2 while other sites in the same culture well remain unaffected. Thus, responses of both hypoxic and bystander cells can be characterised. Moreover, control of microfluidic flow enables generation of intermittent hypoxia or hypoxia/re-oxygenation cycles. STATEMENT OF SIGNIFICANCE: Specific manipulation of oxygen concentrations in cultured cells' microenvironment is important when mimicking low-oxygen tissue conditions and pathologies such as tissue infarction or cancer. We utilised a sodium sulfite-based in-liquid chemical reaction to consume dissolved oxygen. When this liquid was pumped into a microfluidic channel, lowered oxygen levels could be measured outside the channel through a polydimethylsiloxane PDMS membrane allowing only for gaseous exchange. We then utilised this setup to deplete oxygen from the microenvironment of cultured cells, and showed that cells responded to hypoxia on molecular level. Our setup can be used for specifically removing oxygen from the cell culture microenvironment for experimental purposes and for generating a low oxygen environment that better mimics the cells' original tissue environments.


Cell Culture Techniques/methods , Microfluidic Analytical Techniques/methods , Stem Cell Niche , Stem Cells/metabolism , Animals , Cattle , Cell Hypoxia , Stem Cells/cytology
14.
Adv Mater ; 30(24): e1705104, 2018 Jun.
Article En | MEDLINE | ID: mdl-29465772

Superhydrophobic surfaces repel water and, in some cases, other liquids as well. The repellency is caused by topographical features at the nano-/microscale and low surface energy. Blood is a challenging liquid to repel due to its high propensity for activation of intrinsic hemostatic mechanisms, induction of coagulation, and platelet activation upon contact with foreign surfaces. Imbalanced activation of coagulation drives thrombogenesis or formation of blood clots that can occlude the blood flow either on-site or further downstream as emboli, exposing tissues to ischemia and infarction. Blood-repellent superhydrophobic surfaces aim toward reducing the thrombogenicity of surfaces of blood-contacting devices and implants. Several mechanisms that lead to blood repellency are proposed, focusing mainly on platelet antiadhesion. Structured surfaces can: (i) reduce the effective area exposed to platelets, (ii) reduce the adhesion area available to individual platelets, (iii) cause hydrodynamic effects that reduce platelet adhesion, and (iv) reduce or alter protein adsorption in a way that is not conducive to thrombus formation. These mechanisms benefit from the superhydrophobic Cassie state, in which a thin layer of air is trapped between the solid surface and the liquid. The connections between water- and blood repellency are discussed and several recent examples of blood-repellent superhydrophobic surfaces are highlighted.


Hydrophobic and Hydrophilic Interactions , Surface Properties , Water
15.
Sci Rep ; 7(1): 16019, 2017 11 22.
Article En | MEDLINE | ID: mdl-29167540

A top-down scalable method to produce flexible water and blood repellent tubes is introduced. The method is based on replication of overhanging nanostructures from an aluminum tube template to polydimethylsiloxane (PDMS) via atomic layer deposition (ALD) assisted sacrificial etching. The nanostructured PDMS/titania tubes are superhydrophobic with water contact angles 163 ± 1° (advancing) and 157 ± 1° (receding) without any further coating. Droplets are able to slide through a 4 mm (inner diameter) tube with low sliding angles of less than 10° for a 35 µL droplet. The superhydrophobic tube shows up to 5,000 times increase in acceleration of a sliding droplet compared to a control tube depending on the inclination angle. Compared to a free falling droplet, the superhydrophobic tube reduced the acceleration by only 38.55%, as compared to a 99.99% reduction for a control tube. The superhydrophobic tubes are blood repellent. Blood droplets (35 µL) roll through the tubes at 15° sliding angles without leaving a bloodstain. The tube surface is resistant to adhesion of activated platelets unlike planar control titania and smooth PDMS surfaces.


Blood , Nanostructures/chemistry , Water/chemistry , Animals , Dimethylpolysiloxanes/chemistry , Humans , Surface Properties
16.
Chemistry ; 23(66): 16835-16842, 2017 Nov 27.
Article En | MEDLINE | ID: mdl-28902425

Microreactors attract a significant interest for chemical synthesis due to the benefits of small scales such as high surface to volume ratio, rapid thermal ramping, and well-understood laminar flows. The suitability of atomic layer deposition for application of both the nanoparticle catalyst and the support material on the surfaces of channels of microfabricated silicon microreactors is demonstrated in this research. Continuous-flow hydrogenation of propene into propane at low temperatures with TiO2 -supported catalytic Pt nanoparticles was used as a model reaction. Reaction yield and mass transport were monitored by high-sensitivity microcoil NMR spectroscopy as well as time-of-flight remote detection NMR imaging. The microreactors were shown to be very efficient in propene conversion into propane. The yield of 100 % was achieved at 50 °C with a reactor decorated with Pt nanoparticles of average size of roughly 1 nm and surface coverage of 3.2 % in 20 mm long reaction channels with a residence time of 1100 ms. The activity of the Pt catalyst surfaces was on the order of several to tens of mmol s-1 m-2 .

17.
Small ; 13(38)2017 10.
Article En | MEDLINE | ID: mdl-28815888

Reliable characterization of wetting properties is essential for the development and optimization of superhydrophobic surfaces. Here, the dynamics of superhydrophobicity is studied including droplet friction and wetting transitions by using droplet oscillations on micropillared surfaces. Analyzing droplet oscillations by high-speed camera makes it possible to obtain energy dissipation parameters such as contact angle hysteresis force and viscous damping coefficients, which indicate pinning and viscous losses, respectively. It is shown that the dissipative forces increase with increasing solid fraction and magnetic force. For 10 µm diameter pillars, the solid fraction range within which droplet oscillations are possible is between 0.97% and 2.18%. Beyond the upper limit, the oscillations become heavily damped due to high friction force. Below the lower limit, the droplet is no longer supported by the pillar tops and undergoes a Cassie-Wenzel transition. This transition is found to occur at lower pressure for a moving droplet than for a static droplet. The findings can help to optimize micropillared surfaces for low-friction droplet transport.

18.
J Mater Chem B ; 5(45): 9033-9044, 2017 Dec 07.
Article En | MEDLINE | ID: mdl-32264131

Here we investigated the electrochemical properties and dopamine (DA) detection capability of SU-8 photoresist based pyrolytic carbon (PyC) as well as its biocompatibility with neural cells. This approach is compatible with microfabrication techniques which is crucial for device development. X-ray photoelectron spectroscopy shows that PyC consists 98.5% of carbon, while oxygen plasma treatment (PyC-O2) increases the amount of oxygen up to 27.1%. PyC showed nearly reversible (ΔEp 63 mV) electron transfer kinetics towards outer sphere redox probe (Ru(NH3)6 2+/3+), while the reaction on PyC-O2 was quasi-reversible (ΔEp > 75 mV). DA showed both diffusion and adsorption-defined reaction kinetics with fast electron transfer with the ΔEp values of 50 mV and 30 mV, for PyC and PyC-O2, respectively. The strong interaction between the hydroxyl groups on the surface and DA, as confirmed by simulations, facilitates the redox reactions of DA. DA showed a linear response in the measured physiologically relevant range (50 nM-1 µM) and sensitivities were 1.2 A M-1 cm-2 for PyC and 2.7 A M-1 cm-2 for PyC-O2. Plasma oxidation (PyC-O2) improved cell adhesion even more than poly-l-lysine (PLL) coating on PyC, but best adhesion was achieved on PLL coated PyC-O2. Glial cells, neuroblastoma cells and neural stem cells all showed similar behavior.

19.
ACS Appl Mater Interfaces ; 8(24): 15607-14, 2016 Jun 22.
Article En | MEDLINE | ID: mdl-27232271

Self-standing films (45 µm thick) of native cellulose nanofibrils (CNFs) were synthesized and characterized for their piezoelectric response. The surface and the microstructure of the films were evaluated with image-based analysis and scanning electron microscopy (SEM). The measured dielectric properties of the films at 1 kHz and 9.97 GHz indicated a relative permittivity of 3.47 and 3.38 and loss tangent tan Î´ of 0.011 and 0.071, respectively. The films were used as functional sensing layers in piezoelectric sensors with corresponding sensitivities of 4.7-6.4 pC/N in ambient conditions. This piezoelectric response is expected to increase remarkably upon film polarization resulting from the alignment of the cellulose crystalline regions in the film. The CNF sensor characteristics were compared with those of polyvinylidene fluoride (PVDF) as reference piezoelectric polymer. Overall, the results suggest that CNF is a suitable precursor material for disposable piezoelectric sensors, actuators, or energy generators with potential applications in the fields of electronics, sensors, and biomedical diagnostics.


Cellulose/chemistry , Chemistry Techniques, Analytical/instrumentation , Chemistry Techniques, Analytical/methods , Electromagnetic Phenomena , Nanofibers/chemistry , Polymers/chemistry , Surface Properties
20.
Front Cell Neurosci ; 10: 3, 2016.
Article En | MEDLINE | ID: mdl-26834558

Kainate type of glutamate receptors (KARs) are highly expressed during early brain development and may influence refinement of the circuitry, via modulating synaptic transmission and plasticity. KARs are also localized to axons, however, their exact roles in regulating presynaptic processes remain controversial. Here, we have used a microfluidic chamber system allowing specific manipulation of KARs in presynaptic neurons to study their functions in synaptic development and function in vitro. Silencing expression of endogenous KARs resulted in lower density of synaptophysin immunopositive puncta in microfluidically isolated axons. Various recombinant KAR subunits and pharmacological compounds were used to dissect the mechanisms behind this effect. The calcium permeable (Q) variants of the low-affinity (GluK1-3) subunits robustly increased synaptophysin puncta in axons in a manner that was dependent on receptor activity and PKA and PKC dependent signaling. Further, an associated increase in the mean active zone length was observed in electron micrographs. Selective presynaptic expression of these subunits resulted in higher success rate of evoked EPSCs consistent with higher probability of glutamate release. In contrast, the calcium-impermeable (R) variant of GluK1 or the high-affinity subunits (GluK4,5) had no effect on synaptic density or transmission efficacy. These data suggest that calcium permeable axonal KARs promote efferent connectivity by increasing the density of functional presynaptic release sites.

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