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1.
Phys Rev Lett ; 132(14): 146902, 2024 Apr 05.
Article in English | MEDLINE | ID: mdl-38640398

ABSTRACT

Spin defects in silicon carbide are promising candidates for quantum sensing applications as they exhibit long coherence times even at room temperature. However, spin readout methods that rely on fluorescence detection can be challenging due to poor photon collection efficiency. Here, we demonstrate coherent spin control and all-electrical readout of a small ensemble of spins in a SiC junction diode using pulsed electrically detected magnetic resonance. A lock-in detection scheme based on a three stage modulation cycle is implemented, significantly enhancing the signal-to-noise ratio. This technique enabled observation of coherent spin dynamics, specifically Rabi spin nutation, spin dephasing, and spin decoherence. The use of these protocols for magnetometry applications is evaluated.

2.
Phys Rev Lett ; 132(18): 186402, 2024 May 03.
Article in English | MEDLINE | ID: mdl-38759200

ABSTRACT

A gallium interstitial defect is thought to be responsible for the spectacular spin-dependent recombination in GaAs_{1-x}N_{x} dilute nitrides. Current understanding associates this defect with at least two in-gap levels corresponding to the (+/0) and (++/+) charge-state transitions. Using a spin-sensitive photoinduced current transient spectroscopy, the in-gap electronic structure of a x=0.021 alloy is revealed. The (+/0) state lies ≈0.27 eV below the conduction band edge, and an anomalous, negative activation energy reveals the presence of not one but two other in-gap states. The observations are consistent with a (++/+) state ≈0.19 eV above the valence band edge, and a (+++/++) state ≈25 meV above the valence band edge.

3.
Nano Lett ; 19(7): 4543-4550, 2019 Jul 10.
Article in English | MEDLINE | ID: mdl-31150580

ABSTRACT

The precise measurement of mechanical stress at the nanoscale is of fundamental and technological importance. In principle, all six independent variables of the stress tensor, which describe the direction and magnitude of compression/tension and shear stress in a solid, can be exploited to tune or enhance the properties of materials and devices. However, existing techniques to probe the local stress are generally incapable of measuring the entire stress tensor. Here, we make use of an ensemble of atomic-sized in situ strain sensors in diamond (nitrogen-vacancy defects) to achieve spatial mapping of the full stress tensor, with a submicrometer spatial resolution and a sensitivity of the order of 1 MPa (10 MPa) for the shear (axial) stress components. To illustrate the effectiveness and versatility of the technique, we apply it to a broad range of experimental situations, including mapping the stress induced by localized implantation damage, nanoindents, and scratches. In addition, we observe surprisingly large stress contributions from functional electronic devices fabricated on the diamond and also demonstrate sensitivity to deformations of materials in contact with the diamond. Our technique could enable in situ measurements of the mechanical response of diamond nanostructures under various stimuli, with potential applications in strain engineering for diamond-based quantum technologies and in nanomechanical sensing for on-chip mass spectroscopy.

4.
BMC Plant Biol ; 19(1): 172, 2019 Apr 30.
Article in English | MEDLINE | ID: mdl-31039740

ABSTRACT

BACKGROUND: Angiosperm sex chromosomes, where present, are generally recently evolved. The key step in initiating the development of sex chromosomes from autosomes is the establishment of a sex-determining locus within a region of non-recombination. To better understand early sex chromosome evolution, it is important to determine the process by which recombination is suppressed around the sex determining genes. We have used the dioecious angiosperm kiwifruit Actinidia chinensis var. chinensis, which has an active-Y sex chromosome system, to study recombination rates around the sex locus, to better understand key events in the development of sex chromosomes. RESULTS: We have confirmed the sex-determining region (SDR) in A. chinensis var. chinensis, using a combination of high density genetic mapping and fluorescent in situ hybridisation (FISH) of Bacterial Artificial Chromosomes (BACs) linked to the sex markers onto pachytene chromosomes. The SDR is a subtelomeric non-recombining region adjacent to the nucleolar organiser region (NOR). A region of restricted recombination of around 6 Mbp in size in both male and female maps spans the SDR and covers around a third of chromosome 25. CONCLUSIONS: As recombination is suppressed over a similar region between X chromosomes and between and X and Y chromosomes, we propose that recombination is suppressed in this region because of the proximity of the NOR and the centromere, with both the NOR and centromere suppressing recombination, and this predates suppressed recombination due to differences between X and Y chromosomes. Such regions of suppressed recombination in the genome provide an opportunity for the evolution of sex chromosomes, if a sex-determining locus develops there or translocates into this region.


Subject(s)
Actinidia/genetics , Chromosomes, Plant , Recombination, Genetic , Sex Chromosomes , Actinidia/cytology , Chromosome Mapping , Chromosomes, Artificial, Bacterial , Genetic Variation , In Situ Hybridization, Fluorescence , Microsatellite Repeats
5.
Phys Rev Lett ; 122(10): 105701, 2019 Mar 15.
Article in English | MEDLINE | ID: mdl-30932683

ABSTRACT

The rhombohedral phase of Si (r8-Si), a promising semiconducting material, is formed by indentation together with the body-centered cubic phase (bc8-Si). Using a novel sample preparation method, x-ray diffraction is used to determine the relative volume of these phases in indented Si and allow observation of a distorted unit cell along the direction of indentation loading. Theoretical calculations together with these observations suggest the indent contains an intrinsic compression of ∼4 GPa that stabilizes the r8 phase.

6.
Rep Prog Phys ; 80(3): 034502, 2017 03.
Article in English | MEDLINE | ID: mdl-28139468

ABSTRACT

This paper summarizes key findings in single-photon generation from deep level defects in silicon carbide (SiC) and highlights the significance of these individually addressable centers for emerging quantum applications. Single photon emission from various defect centers in both bulk and nanostructured SiC are discussed as well as their formation and possible integration into optical and electrical devices. The related measurement protocols, the building blocks of quantum communication and computation network architectures in solid state systems, are also summarized. This includes experimental methodologies developed for spin control of different paramagnetic defects, including the measurement of spin coherence times. Well established doping, and micro- and nanofabrication procedures for SiC may allow the quantum properties of paramagnetic defects to be electrically and mechanically controlled efficiently. The integration of single defects into SiC devices is crucial for applications in quantum technologies and we will review progress in this direction.

7.
Acta Chir Orthop Traumatol Cech ; 84(3): 211-214, 2017.
Article in English | MEDLINE | ID: mdl-28809642

ABSTRACT

Posttraumatic pelvic nonunions in combination with malposition are uncommon in the present-day era of modern pelvic surgery. The case describes a new surgical technique for treatment of the nonunion localized to iliosacral joint. A 42-year-old polytraumatized male presented with a pelvic fracture (type 61-C2.3) associated with a complex acetabular fracture. The patient was treated and the pelvis stabilized according to damage control principles (external fixation and pelvic C-clamp) and subsequently definitively treated according to principles of current pelvic surgery. The posterior pelvis was stabilized by bilateral S1 iliosacral screws, however the posterior pelvic ring injury on the left side was not well reduced. The result was a nonunion formation in the left sacroiliacal joit with screw loosening. Two attemps at bone grafting and repeat stabilization were done, but the nonunion did not heal. The end result was a nonunion through left sacroiliacal joint with destruction of the lateral part of sacral bone. The nonunion was treated with nonvascularised autologous fibular strut graft in combination with allogenic corticocancellous grafting. The fibular graft was placed into the bone void after the removal of the iliosacral screw. Radiographically the nonunion healed completely six months after the fibular grafting and the patient had improved clinical outcome. The described technique solves both instability and bone defect in posterior pelvic ring even in the case with low contact areas. The nonvascularised autologous fibular grafting is an effective technique for sacroiliac joint nonunion treatment even in case with large bone defect. Key words: pelvis; nonunion; fibular graft.


Subject(s)
Acetabulum/injuries , Bone Transplantation , Fibula/transplantation , Fracture Fixation, Internal , Fractures, Ununited/surgery , Pelvic Bones/injuries , Adult , Humans , Male
8.
J Neurosci ; 32(34): 11737-49, 2012 Aug 22.
Article in English | MEDLINE | ID: mdl-22915116

ABSTRACT

Currents through voltage-gated Ca²âº channels (I(Ca)) may be regulated by cytoplasmic Ca²âº levels ([Ca²âº](c)), producing Ca²âº-dependent inactivation (CDI) or facilitation (CDF). Since I(Ca) regulates sensory neuron excitability, altered CDI or CDF could contribute to pain generation after peripheral nerve injury. We explored this by manipulating [Ca²âº](c) while recording I(Ca) in rat sensory neurons. In uninjured neurons, elevating [Ca²âº](c) with a conditioning prepulse (-15 mV, 2 s) inactivated I(Ca) measured during subsequent test pulses (-15 mV, 5 ms). This inactivation was Ca²âº-dependent (CDI), since it was decreased with elimination of Ca²âº influx by depolarization to above the I(Ca) reversal potential, with high intracellular Ca²âº buffering (EGTA 10 mm or BAPTA 20 mm), and with substitution of Ba²âº for extracellular Ca²âº, revealing a residual voltage-dependent inactivation. At longer latencies after conditioning (>6 s), I(Ca) recovered beyond baseline. This facilitation also proved to be Ca²âº-dependent (CDF) using the protocols limiting cytoplasmic Ca²âº elevation. Ca²âº/calmodulin-dependent protein kinase II (CaMKII) blockers applied by bath (KN-93, myristoyl-AIP) or expressed selectively in the sensory neurons (AIP) reduced CDF, unlike their inactive analogues. Protein kinase C inhibition (chelerythrine) had no effect. Selective blockade of N-type Ca²âº channels eliminated CDF, whereas L-type channel blockade had no effect. Following nerve injury, CDI was unaffected, but CDF was eliminated in axotomized neurons. Excitability of sensory neurons in intact ganglia from control animals was diminished after a similar conditioning pulse, but this regulation was eliminated by injury. These findings indicate that I(Ca) in sensory neurons is subject to both CDI and CDF, and that hyperexcitability following injury-induced loss of CDF may result from diminished CaMKII activity.


Subject(s)
Biophysical Phenomena/physiology , Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism , Calcium/metabolism , Neurons, Afferent/physiology , Peripheral Nerve Injuries/pathology , Signal Transduction/physiology , Analysis of Variance , Animals , Biophysical Phenomena/drug effects , Biophysics , Calcium Channel Blockers/pharmacology , Calcium Signaling/drug effects , Chelating Agents/pharmacology , Dantrolene/pharmacology , Drug Interactions , Egtazic Acid/analogs & derivatives , Electric Stimulation , Enzyme Inhibitors/pharmacology , Ganglia, Spinal/cytology , Genetic Vectors/physiology , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Hyperalgesia/etiology , Hyperalgesia/metabolism , Laminectomy , Male , Membrane Potentials/drug effects , Neurons, Afferent/drug effects , Pain Threshold/drug effects , Patch-Clamp Techniques , Peripheral Nerve Injuries/complications , Peripheral Nerve Injuries/enzymology , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects , Time Factors
9.
Anesthesiology ; 118(2): 350-60, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23287707

ABSTRACT

INTRODUCTION: Previously observed increased sensitivity to noxious stimulation in the Dahl salt-sensitive rat strain (SS/JrHsdMcwi, abbreviated as SS) compared to Brown Norway rats (BN/NhsdMcwi abbreviated as BN) is mediated by genes on a single chromosome. The current study used behavioral and electrocortical data to determine if differences also exist between SS and BN rats in loss of consciousness. METHODS: Behavioral responses, including loss of righting, (a putative index of consciousness) and concurrent electroencephalogram recordings, in 12 SS and BN rats were measured during isoflurane at inhaled concentrations of 0, 0.3, 0.6, 0.8, 1.0 and 1.2%. RESULTS: In SS compared to BN rats, the mean ± SEM EC50 for righting was significantly less (0.65 ± 0.01% vs. 0.74 ± 0.02% inhaled isoflurane) and delta fraction in parietal electroencephalogram was enhanced 50-100% at all isoflurane levels during emergence. The frequency decay constant of an exponential fit of the parietal electroencephalogram spectrum graphed as a function of isoflurane level was three times less steep (mean ± SEM slope -57 ± 13 vs. -191 ± 38) and lower at each level of isoflurane in SS versus BN rats (i.e., shifted toward low frequency activity). Electroencephalogram differences between strains were larger during emergence than induction. CONCLUSIONS: Sensitivity is higher in SS compared to BN rats leading to unconsciousness at lower levels of isoflurane. This supports using additional strains in this animal model to study the genetic basis for differences in anesthetic action on mechanisms of consciousness. Moreover, induction and emergence appear to involve distinct pathways.


Subject(s)
Anesthesia, Inhalation , Anesthetics, Inhalation , Electroencephalography , Isoflurane , Unconsciousness/chemically induced , Unconsciousness/genetics , Algorithms , Anesthetics, Inhalation/blood , Animals , Behavior, Animal/drug effects , Dose-Response Relationship, Drug , Electrodes, Implanted , Isoflurane/blood , Male , Rats , Rats, Inbred BN , Rats, Inbred Dahl , Species Specificity
10.
Nanotechnology ; 24(14): 145304, 2013 Apr 12.
Article in English | MEDLINE | ID: mdl-23508018

ABSTRACT

Solid state electronic devices fabricated in silicon employ many ion implantation steps in their fabrication. In nanoscale devices deterministic implants of dopant atoms with high spatial precision will be needed to overcome problems with statistical variations in device characteristics and to open new functionalities based on controlled quantum states of single atoms. However, to deterministically place a dopant atom with the required precision is a significant technological challenge. Here we address this challenge with a strategy based on stepped nanostencil lithography for the construction of arrays of single implanted atoms. We address the limit on spatial precision imposed by ion straggling in the nanostencil-fabricated with the readily available focused ion beam milling technique followed by Pt deposition. Two nanostencils have been fabricated; a 60 nm wide aperture in a 3 µm thick Si cantilever and a 30 nm wide aperture in a 200 nm thick Si3N4 membrane. The 30 nm wide aperture demonstrates the fabricating process for sub-50 nm apertures while the 60 nm aperture was characterized with 500 keV He(+) ion forward scattering to measure the effect of ion straggling in the collimator and deduce a model for its internal structure using the GEANT4 ion transport code. This model is then applied to simulate collimation of a 14 keV P(+) ion beam in a 200 nm thick Si3N4 membrane nanostencil suitable for the implantation of donors in silicon. We simulate collimating apertures with widths in the range of 10-50 nm because we expect the onset of J-coupling in a device with 30 nm donor spacing. We find that straggling in the nanostencil produces mis-located implanted ions with a probability between 0.001 and 0.08 depending on the internal collimator profile and the alignment with the beam direction. This result is favourable for the rapid prototyping of a proof-of-principle device containing multiple deterministically implanted dopants.

11.
Proc Natl Acad Sci U S A ; 106(21): 8725-30, 2009 May 26.
Article in English | MEDLINE | ID: mdl-19439665

ABSTRACT

Painful axotomy decreases K(ATP) channel current (IK(ATP)) in primary afferent neurons. Because cytosolic Ca(2+) signaling is depressed in injured dorsal root ganglia (DRG) neurons, we investigated whether Ca(2+)-calmodulin (CaM)-Ca(2+)/CaM-dependent kinase II (CaMKII) regulates IK(ATP) in large DRG neurons. Immunohistochemistry identified the presence of K(ATP) channel subunits SUR1, SUR2, and Kir6.2 but not Kir6.1, and pCaMKII in neurofilament 200-positive DRG somata. Single-channel recordings from cell-attached patches revealed that basal and evoked IK(ATP) by ionomycin, a Ca(2+) ionophore, is activated by CaMKII. In axotomized neurons from rats made hyperalgesic by spinal nerve ligation (SNL), basal K(ATP) channel activity was decreased, and sensitivity to ionomycin was abolished. Basal and Ca(2+)-evoked K(ATP) channel activity correlated inversely with the degree of hyperalgesia induced by SNL in the rats from which the neurons were isolated. Inhibition of IK(ATP) by glybenclamide, a selective K(ATP) channel inhibitor, depolarized resting membrane potential (RMP) recorded in perforated whole-cell patches and enhanced neurotransmitter release measured by amperometry. The selective K(ATP) channel opener diazoxide hyperpolarized the RMP and attenuated neurotransmitter release. Axotomized neurons from rats made hyperalgesic by SNL lost sensitivity to the myristoylated form of autocamtide-2-related inhibitory peptide (AIPm), a pseudosubstrate blocker of CaMKII, whereas axotomized neurons from SNL animals that failed to develop hyperalgesia showed normal IK(ATP) inhibition by AIPm. AIPm also depolarized RMP in control neurons via K(ATP) channel inhibition. Unitary current conductance and sensitivity of K(ATP) channels to cytosolic ATP and ligands were preserved even after painful nerve injury, thus providing opportunities for selective therapeutic targeting against neuropathic pain.


Subject(s)
Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism , Calcium/metabolism , Calmodulin/metabolism , Hyperalgesia/metabolism , KATP Channels/metabolism , Neurons, Afferent/metabolism , Animals , Axotomy , Cell-Free System , Electrophysiological Phenomena , Ganglia, Spinal/metabolism , Ionomycin/pharmacology , Male , Patch-Clamp Techniques , Rats , Rats, Sprague-Dawley
12.
Anesthesiology ; 115(4): 733-42, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21804378

ABSTRACT

BACKGROUND: The nucleus basalis of Meynert of the basal forebrain has been implicated in the regulation of the state of consciousness across normal sleep-wake cycles. Its role in the modulation of general anesthesia was investigated. METHODS: Rats were chronically implanted with bilateral infusion cannulae in the nucleus basalis of Meynert and epidural electrodes to record the electroencephalogram in frontal and visual cortices. Animals were anesthetized with desflurane at a concentration required for the loss of righting reflex (4.6 ± 0.5%). Norepinephrine (17.8 nmol) or artificial cerebrospinal fluid was infused at 0.2 µl/min (1 µl total). Behavioral response to infusion was measured by scoring the orofacial, limb, and head movements, and postural changes. RESULTS: Behavioral responses were higher after norepinephrine (2.1 ± 1) than artificial cerebrospinal fluid (0.63 ± 0.8) infusion (P < 0.01, Student t test). Responses were brief (1-2 min), repetitive, and more frequent after norepinephrine infusion (P < 0.0001, chi-square test). Electroencephalogram delta power decreased after norepinephrine in frontal (70 ± 7%) but not in visual cortex (P < 0.05, Student t test). Simultaneously, electroencephalogram cross-approximate entropy between frontal and visual cortices increased from 3.17 ± 0.56 to 3.85 ± 0.29 after norepinephrine infusion (P < 0.01, Student t test). Behavioral activation was predictable by the decrease in frontal delta power (logistic regression, P < 0.05). CONCLUSIONS: Norepinephrine infusion into the nucleus basalis of Meynert can modulate anesthetic depth presumably by ascending activation of the cortex. The transient nature of the responses suggests a similarity with microarousals normally observed during natural sleep, and may imply a mechanism for transient awareness under light anesthesia.


Subject(s)
Adrenergic alpha-Agonists/administration & dosage , Adrenergic alpha-Agonists/pharmacology , Anesthetics, Inhalation/pharmacology , Arousal/drug effects , Basal Nucleus of Meynert/physiology , Isoflurane/analogs & derivatives , Norepinephrine/administration & dosage , Norepinephrine/pharmacology , Anesthesia, Inhalation , Animals , Behavior, Animal/drug effects , Consciousness/drug effects , Data Interpretation, Statistical , Desflurane , Electroencephalography/drug effects , Entropy , Injections , Isoflurane/pharmacology , Male , Motor Activity/drug effects , Prefrontal Cortex/drug effects , Rats , Rats, Sprague-Dawley , Reflex, Righting , Sleep/drug effects , Visual Cortex/drug effects
13.
Intern Med J ; 41(7): 555-60, 2011 Jul.
Article in English | MEDLINE | ID: mdl-19849748

ABSTRACT

BACKGROUND: This study examines the predictors of long-term all-causes mortality (ACM) in Australian senior citizens. METHODS: We have analysed ACM in a cohort of 2805 citizens, 1233 men and 1572 women aged ≥60 years, first examined in 1988 and followed for 20 years. Hazard ratios and 95% confidence intervals for ACM were obtained from Cox models employing conventional predictors. RESULTS: Over 20 years 66% of men (815/1233) and 53% of women (833/1572) died. Constant proportional hazard over the 20 years was demonstrated for all predictors, indicating similar relative hazard of ACM during long-term or short-term follow up. There was significant prediction of ACM by current smoking (hazard ratio 1.96, 95% confidence interval 1.57-2.43 in men; 1.67, 1.32-2.10 in women), high blood pressure (1.37, 1.03-1.81; 1.41, 1.07-1.86), diabetes (1.46, 1.17-1.82; 1.83, 1.43-2.34), impaired peak expiratory flow (1.39, 1.15-1.69; 1.80, 1.47-2.21), coronary heart disease at study entry in men (1.33, 1.13-1.57), physical disability (1.38, 1.13-1.68; 1.45, 1.17-1.79) and alcohol intake (0.82, 0.69-0.97; 0.77, 0.66-0.89 respectively). ACM was not significantly predicted by standard lipid parameters. Over the 20-year period smoking was associated with reduced survival of 41 months in men and 25 months in women, hypertension with reduced survival of 20 and 17 months, and diabetes with reduced survival of 24 and 30 months respectively. CONCLUSIONS: The findings confirm the contribution of cigarette smoking, hypertension and diabetes to ACM in senior citizens, conditions that are potentially amenable to intervention.


Subject(s)
Diabetes Mellitus/mortality , Hypertension/mortality , Smoking/mortality , Age Factors , Aged , Aged, 80 and over , Female , Follow-Up Studies , Humans , Male , Middle Aged , New South Wales/epidemiology , Predictive Value of Tests , Risk Factors , Sex Factors , Time Factors
14.
J Insect Sci ; 11: 98, 2011.
Article in English | MEDLINE | ID: mdl-22208869

ABSTRACT

Onion, Allium cepa L. (Asparagales: Amaryllidaceae), crop fields grown for seed production require arthropod pollination for adequate seed yield. Although many arthropod species visit A. cepa flowers, for most there is little information on their role as pollinators. Small flower visiting arthropods (body width < 3 mm) in particular are rarely assessed. A survey of eight flowering commercial A. cepa seed fields in the North and South Islands of New Zealand using window traps revealed that small arthropods were highly abundant among all except one field. Insects belonging to the orders Diptera and Thysanoptera were the most abundant and Hymenoptera, Collembola, Psocoptera, Hemiptera, and Coleoptera were also present. To test whether small arthropods might contribute to pollination, seed sets from umbels caged within 3 mm diameter mesh cages were compared with similarly caged, hand-pollinated umbels and uncaged umbels. Caged umbels that were not hand-pollinated set significantly fewer seeds (average eight seeds/umbel, n = 10) than caged hand-pollinated umbels (average 146 seeds/umbel) and uncaged umbels (average 481 seeds/umbel). Moreover, sticky traps placed on umbels within cages captured similar numbers of small arthropods as sticky traps placed on uncaged umbels, suggesting cages did not inhibit the movement of small arthropods to umbels. Therefore, despite the high abundance of small arthropods within fields, evidence to support their role as significant pollinators of commercial A. cepa seed crops was not found.


Subject(s)
Allium , Arthropods , Biodiversity , Pollination , Animals , Crops, Agricultural , Flowers , New Zealand , Seeds
15.
Mol Pain ; 5: 12, 2009 Mar 14.
Article in English | MEDLINE | ID: mdl-19284878

ABSTRACT

BACKGROUND: ATP-sensitive potassium (KATP) channels in neurons regulate excitability, neurotransmitter release and mediate protection from cell-death. Furthermore, activation of KATP channels is suppressed in DRG neurons after painful-like nerve injury. NO-dependent mechanisms modulate both KATP channels and participate in the pathophysiology and pharmacology of neuropathic pain. Therefore, we investigated NO modulation of KATP channels in control and axotomized DRG neurons. RESULTS: Cell-attached and cell-free recordings of KATP currents in large DRG neurons from control rats (sham surgery, SS) revealed activation of KATP channels by NO exogenously released by the NO donor SNAP, through decreased sensitivity to [ATP]i. This NO-induced KATP channel activation was not altered in ganglia from animals that demonstrated sustained hyperalgesia-type response to nociceptive stimulation following spinal nerve ligation. However, baseline opening of KATP channels and their activation induced by metabolic inhibition was suppressed by axotomy. Failure to block the NO-mediated amplification of KATP currents with specific inhibitors of sGC and PKG indicated that the classical sGC/cGMP/PKG signaling pathway was not involved in the activation by SNAP. NO-induced activation of KATP channels remained intact in cell-free patches, was reversed by DTT, a thiol-reducing agent, and prevented by NEM, a thiol-alkylating agent. Other findings indicated that the mechanisms by which NO activates KATP channels involve direct S-nitrosylation of cysteine residues in the SUR1 subunit. Specifically, current through recombinant wild-type SUR1/Kir6.2 channels expressed in COS7 cells was activated by NO, but channels formed only from truncated isoform Kir6.2 subunits without SUR1 subunits were insensitive to NO. Further, mutagenesis of SUR1 indicated that NO-induced KATP channel activation involves interaction of NO with residues in the NBD1 of the SUR1 subunit. CONCLUSION: NO activates KATP channels in large DRG neurons via direct S-nitrosylation of cysteine residues in the SUR1 subunit. The capacity of NO to activate KATP channels via this mechanism remains intact even after spinal nerve ligation, thus providing opportunities for selective pharmacological enhancement of KATP current even after decrease of this current by painful-like nerve injury.


Subject(s)
Ion Channel Gating/drug effects , KATP Channels/metabolism , Mammals/metabolism , Nitric Oxide/pharmacology , Sensory Receptor Cells/drug effects , Sensory Receptor Cells/metabolism , ATP-Binding Cassette Transporters/chemistry , ATP-Binding Cassette Transporters/metabolism , Animals , COS Cells , Chlorocebus aethiops , Cyclic GMP/analogs & derivatives , Cyclic GMP/pharmacology , Cyclic GMP-Dependent Protein Kinases/metabolism , Cysteine/genetics , Ganglia, Spinal/cytology , Ganglia, Spinal/metabolism , Male , Mutation/genetics , Nitric Oxide/metabolism , Nitrosation/drug effects , Patch-Clamp Techniques , Potassium Channels, Inwardly Rectifying/chemistry , Potassium Channels, Inwardly Rectifying/metabolism , Protein Structure, Tertiary , Rats , Receptors, Drug/chemistry , Receptors, Drug/metabolism , Recombinant Proteins/metabolism , S-Nitroso-N-Acetylpenicillamine/pharmacology , Sensory Receptor Cells/enzymology , Sulfonylurea Receptors
16.
Neuron ; 28(3): 887-98, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11163274

ABSTRACT

The NMDA receptor NR1 subunit has four splice variants that differ in their C-terminal, cytoplasmic domain. We investigated the contribution of the C-terminal cassettes, C0, C1, C2, and C2', to trafficking of NR1 in heterologous cells and neurons. We identified an ER retention signal (RRR) in the C1 cassette of NR1, which is similar to the RXR motif in ATP-sensitive K(+) channels (Zerangue et al., 1999). We found that surface expression of NR1-3, which contains C1, is due to a site on the C2' cassette, which includes the terminal 4 amino acid PDZ-interacting domain. This site suppresses ER retention of the C1 cassette and leads to surface expression. These findings suggest a role for PDZ proteins in facilitating the transition of receptors from an intracellular pool to the surface of the neuron.


Subject(s)
Alternative Splicing/genetics , Endoplasmic Reticulum/metabolism , Neurons/metabolism , Receptors, N-Methyl-D-Aspartate/genetics , Receptors, N-Methyl-D-Aspartate/metabolism , Amino Acid Motifs/genetics , Animals , Cell Membrane/metabolism , Cells, Cultured , HeLa Cells , Humans , Nerve Tissue Proteins/metabolism , Neurons/cytology , Protein Binding/genetics , Protein Structure, Tertiary/genetics , Rats , Rats, Sprague-Dawley , Signal Transduction/physiology
17.
Nat Neurosci ; 4(8): 794-802, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11477425

ABSTRACT

Although synaptic AMPA receptors have been shown to rapidly internalize, synaptic NMDA receptors are reported to be static. It is not certain whether NMDA receptor stability at synaptic sites is an inherent property of the receptor, or is due to stabilization by scaffolding proteins. In this study, we demonstrate that NMDA receptors are internalized in both heterologous cells and neurons, and we define an internalization motif, YEKL, on the distal C-terminus of NR2B. In addition, we show that the synaptic protein PSD-95 inhibits NR2B-mediated internalization, and that deletion of the PDZ-binding domain of NR2B increases internalization in neurons. This suggests an involvement for PSD-95 in NMDA receptor regulation and an explanation for NMDA receptor stability at synaptic sites.


Subject(s)
Central Nervous System/metabolism , Endocytosis/physiology , Neurons/metabolism , Protein Transport/physiology , Receptors, N-Methyl-D-Aspartate/chemistry , Receptors, N-Methyl-D-Aspartate/metabolism , Synaptic Membranes/metabolism , Amino Acid Motifs/physiology , Animals , Binding Sites/physiology , Central Nervous System/ultrastructure , Clathrin/metabolism , Disks Large Homolog 4 Protein , Fetus , HeLa Cells/cytology , HeLa Cells/metabolism , Hippocampus/cytology , Hippocampus/metabolism , Humans , Immunohistochemistry , Intracellular Signaling Peptides and Proteins , Membrane Proteins , Nerve Tissue Proteins/metabolism , Neurons/ultrastructure , Protein Structure, Tertiary/physiology , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/ultrastructure , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Synaptic Membranes/ultrastructure
18.
Brain Res ; 1132(1): 84-99, 2007 Feb 09.
Article in English | MEDLINE | ID: mdl-17184741

ABSTRACT

UNLABELLED: Calcium-activated potassium channels regulate AHP and excitability in neurons. Since we have previously shown that axotomy decreases I(Ca) in DRG neurons, we investigated the association between I(Ca) and K((Ca)) currents in control medium-sized (30-39 microM) neurons, as well as axotomized L5 or adjacent L4 DRG neurons from hyperalgesic rats following L5 SNL. Currents in response to AP waveform voltage commands were recorded first in Tyrode's solution and sequentially after: 1) blocking Na(+) current with NMDG and TTX; 2) addition of K((Ca)) blockers with a combination of apamin 1 microM, iberiotoxin 200 nM, and clotrimazole 500 nM; 3) blocking remaining K(+) current with the addition of 4-AP, TEA-Cl, and glibenclamide; and 4) blocking I(Ca) with cadmium. In separate experiments, currents were evoked (HP -60 mV, 200 ms square command pulses from -100 to +50 mV) while ensuring high levels of activation of I(K(Ca)) by clamping cytosolic Ca(2+) concentration with pipette solution in which Ca(2+) was buffered to 1 microM. This revealed I(K(Ca)) with components sensitive to apamin, clotrimazole and iberiotoxin. SNL decreases total I(K(Ca)) in axotomized (L5) neurons, but increases total I(K(Ca)) in adjacent (L4) DRG neurons. All I(K(Ca)) subtypes are decreased by axotomy, but iberiotoxin-sensitive and clotrimazole-sensitive current densities are increased in adjacent L4 neurons after SNL. In an additional set of experiments we found that small-sized control DRG neurons also expressed iberiotoxin-sensitive currents, which are reduced in both axotomized (L5) and adjacent (L4) neurons. CONCLUSIONS: Axotomy decreases I(K(Ca)) due to a direct effect on K((Ca)) channels. Axotomy-induced loss of I(Ca) may further potentiate current reduction. This reduction in I(K(Ca)) may contribute to elevated excitability after axotomy. Adjacent neurons (L4 after SNL) exhibit increased I(K(Ca)) current.


Subject(s)
Ganglia, Spinal/metabolism , Neurons, Afferent/metabolism , Peripheral Nervous System Diseases/metabolism , Potassium Channels, Calcium-Activated/metabolism , Sciatic Neuropathy/metabolism , Animals , Axotomy , Calcium/metabolism , Calcium Signaling/drug effects , Calcium Signaling/physiology , Cell Size/drug effects , Cells, Cultured , Ganglia, Spinal/cytology , Ganglia, Spinal/physiopathology , Hyperalgesia/metabolism , Hyperalgesia/physiopathology , Ligation , Male , Membrane Potentials/drug effects , Membrane Potentials/physiology , Neurons, Afferent/cytology , Neurons, Afferent/drug effects , Organ Culture Techniques , Patch-Clamp Techniques , Peripheral Nervous System Diseases/physiopathology , Potassium Channel Blockers/pharmacology , Potassium Channels, Calcium-Activated/drug effects , Rats , Rats, Sprague-Dawley , Sciatic Neuropathy/physiopathology , Sodium Channel Blockers/pharmacology
19.
J Geod ; 91(7): 803-817, 2017.
Article in English | MEDLINE | ID: mdl-32025105

ABSTRACT

The AUSTRAL observing program was started in 2011, performing geodetic and astrometric very long baseline interferometry (VLBI) sessions using the new Australian AuScope VLBI antennas at Hobart, Katherine, and Yarragadee, with contribution from the Warkworth (New Zealand) 12 m and Hartebeesthoek (South Africa) 15 m antennas to make a southern hemisphere array of telescopes with similar design and capability. Designed in the style of the next-generation VLBI system, these small and fast antennas allow for a new way of observing, comprising higher data rates and more observations than the standard observing sessions coordinated by the International VLBI Service for Geodesy and Astrometry (IVS). In this contribution, the continuous development of the AUSTRAL sessions is described, leading to an improvement of the results in terms of baseline length repeatabilities by a factor of two since the start of this program. The focus is on the scheduling strategy and increased number of observations, aspects of automated operation, and data logistics, as well as results of the 151 AUSTRAL sessions performed so far. The high number of the AUSTRAL sessions makes them an important contributor to VLBI end-products, such as the terrestrial and celestial reference frames and Earth orientation parameters. We compare AUSTRAL results with other IVS sessions and discuss their suitability for the determination of baselines, station coordinates, source coordinates, and Earth orientation parameters.

20.
J Neurosci ; 21(19): 7506-16, 2001 Oct 01.
Article in English | MEDLINE | ID: mdl-11567040

ABSTRACT

The regulation of AMPA receptors at the postsynaptic membrane is a fundamental component of synaptic plasticity. In the hippocampus, the induction of long-term potentiation increases the delivery of GluR1, a major AMPA receptor subunit in hippocampal pyramidal neurons, to the synaptic plasma membrane through a mechanism that requires the PDZ binding domain of GluR1. Synapse-associated protein 97 (SAP97), a member of the membrane-associated guanylate kinase family, is believed to associate with AMPA receptors (AMPARs) containing the GluR1 subunit, but the functional significance of these interactions is unclear. We investigated the interaction of GluR1 with SAP97, the only PDZ protein known to interact with GluR1. We find that interactions involving SAP97 and GluR1 occur early in the secretory pathway, while the receptors are in the endoplasmic reticulum or cis-Golgi. In contrast, few synaptic receptors associate with SAP97, suggesting that SAP97 dissociates from the receptor complex at the plasma membrane. We also show that internalization of GluR1, as triggered by NMDAR activation, does not require SAP97. These results implicate GluR1-SAP97 interactions in mechanisms underlying AMPA receptor targeting.


Subject(s)
Nerve Tissue Proteins/metabolism , Receptors, AMPA/metabolism , Adaptor Proteins, Signal Transducing , Animals , Biotinylation , Cell Membrane/chemistry , Cell Membrane/metabolism , Cells, Cultured , Cerebral Cortex/chemistry , Cerebral Cortex/cytology , Cerebral Cortex/metabolism , Cytoplasm/metabolism , Cytoplasm/ultrastructure , Endoplasmic Reticulum/metabolism , Golgi Apparatus/metabolism , Hippocampus/chemistry , Hippocampus/cytology , Hippocampus/metabolism , Immunohistochemistry , Membrane Proteins , Models, Neurological , Neurons/metabolism , Neurons/ultrastructure , Precipitin Tests , Rats , Rats, Sprague-Dawley , Subcellular Fractions/metabolism , Synapses/metabolism , Synapses/ultrastructure
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