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1.
Nano Lett ; 20(12): 8689-8695, 2020 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-33175553

RESUMO

Single-photon emitting point defects in semiconductors have emerged as strong candidates for future quantum technology devices. In the present work, we exploit crystalline particles to investigate relevant defect localizations, emission shifting, and waveguiding. Specifically, emission from 6H-SiC micro- and nanoparticles ranging from 100 nm to 5 µm in size is collected using cathodoluminescence (CL), and we monitor signals attributed to the Si vacancy (VSi) as a function of its location. Clear shifts in the emission wavelength are found for emitters localized in the particle center and at the edges. By comparing spatial CL maps with strain analysis carried out in transmission electron microscopy, we attribute the emission shifts to compressive strain of 2-3% along the particle a-direction. Thus, embedding VSi qubit defects within SiC nanoparticles offers an interesting and versatile opportunity to tune single-photon emission energies while simultaneously ensuring ease of addressability via a self-assembled SiC nanoparticle matrix.

2.
J Phys Condens Matter ; 32(50)2020 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-32756022

RESUMO

The evolution of electrical resistance as function of defect concentration is examined for the unipolarn-conducting oxides CdO,ß-Ga2O3, In2O3, SnO2and ZnO in order to explore the predictions of the amphoteric defect model. Intrinsic defects are introduced by ion irradiation at cryogenic temperatures, and the resistance is measured in-situ by current-voltage sweeps as a function of irradiation dose. Temperature dependent Hall effect measurements are performed to determine the carrier concentration and mobility of the samples before and after irradiation. After the ultimate irradiation step, the Ga2O3and SnO2samples have both turned highly resistive. In contrast, the In2O3and ZnO samples are ultimately found to be less resistive than prior to irradiation, however, they both show an increased resistance at intermediate doses. Based on thermodynamic defect charge state transitions computed by hybrid density functional theory, a model expanding on the current amphoteric defect model is proposed.

3.
Nanoscale Adv ; 2(2): 724-733, 2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-36133232

RESUMO

The optical properties of single ion tracks have been studied in ZnO implanted with Ge by combining depth-resolved hyperspectral cathodoluminescence (CL) and photoluminescence (PL) spectroscopy techniques. The results indicate that ZnO is susceptible to implantation doses as low as 108 to 109 cm-2. We demonstrate that the intensity ratio of ionized and neutral donor bound exciton emissions [D+X/D0X] can be used as a tracer for a local band bending both at the surface as well as in the crystal bulk along the ion tracks. The hyperspectral CL imaging performed at 80 K with 50 nm resolution over the regions with single ion tracks permitted direct assessment of the minority carrier diffusion length. The radii of distortion and space charge surrounding single ion tracks were estimated from the 2D distributions of defect-related green emission (GE) and excitonic D+X emission, both normalized with regard to neutral D0X emission, i.e., from the [GE/D0X] and [D+X/D0X] ratio maps. Our results indicate that single ion tracks in ZnO can be resolved up to ion doses of the order of 5 × 109 cm-2, in which defect aggregation along the extended defects obstructs signatures of individual tracks.

4.
Nanotechnology ; 29(31): 318002, 2018 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-29749345

RESUMO

We respond to the comment by Thomas Walther and reaffirm the findings of our original article.

5.
Nanotechnology ; 28(10): 105703, 2017 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-28085004

RESUMO

Using monochromated electron energy loss spectroscopy in a probe-corrected scanning transmission electron microscope we demonstrate band gap mapping in ZnO/ZnCdO thin films with a spatial resolution below 10 nm and spectral precision of 20 meV.

6.
Nanoscale ; 8(14): 7631-7, 2016 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-26987850

RESUMO

The spatial distribution of defect related deep band emission has been studied in zinc oxide (ZnO) nano- and microwires using depth resolved cathodoluminescence spectroscopy (DRCLS) in a hyperspectral imaging (HSI) mode within a UHV scanning electron microscope (SEM). Three sets of wires were examined that had been grown by pulsed laser deposition or vapor transport methods and ranged in diameter from 200 nm-2.7 µm. This data was analyzed by developing a 3D DRCLS simulation and using it to estimate the segregation depth and decay profile of the near surface defects. We observed different dominant defects from each growth process as well as diameter-dependent defect segregation behavior.

7.
Parasitol Res ; 113(1): 387-90, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24233408

RESUMO

The objective of this study was to determine if human genotypes of Giardia lamblia could be found in canine companion animals from urban and peri-urban environments in Tucson, Arizona. Canine fecal samples collected from the Humane Society of Southern Arizona between July 2006 and April 2009 were screened for G. lamblia infection using immunofluorescent microscopy and confirmed by polymerase chain reaction (PCR). Of the 672 samples screened, 196 were found positive by IFA and 185 of those positive were successfully amplified through PCR. Sequencing analysis showed samples were primarily of the C or D genotypes (n =154), or showing a mix of the C and D genotypes (n =10). One sample showed a mixed infection of a human genotype (A) and a dog-specific genotype (C). These data are consistent with previous studies showing dog specific genotypes to be dominant in environments where dog-to-dog transmission is likely to occur, and provides further evidence that multiple genes should be targeted for more accurate genotype characterization.


Assuntos
Doenças do Cão/parasitologia , Cães/parasitologia , Giardia lamblia/classificação , Giardíase/veterinária , Animais , Arizona/epidemiologia , Doenças do Cão/epidemiologia , Fezes/parasitologia , Genótipo , Giardia lamblia/genética , Giardíase/epidemiologia , Humanos , Reação em Cadeia da Polimerase , Análise de Sequência de DNA
8.
Biochim Biophys Acta ; 1519(3): 246-9, 2001 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-11418193

RESUMO

The monoclonal antibody Lan3-15 identifies a novel protein, Hillarin, that is localized to the axon hillock of leech neurons. Using this antibody we have identified a full length cDNA coding for leech Hillarin and determined its sequence. The gene encodes a 1274 residue protein with a predicted molecular mass of 144013 Da. Data base searches revealed that leech Hillarin has potential orthologues in fly and nematode and that these proteins share two novel protein domains. The W180 domain is characterized by five conserved tryptophans whereas the H domains share 21 invariant residues. In contrast to the arrangement in fly and nematode the cassette containing the W180 and H domains is repeated twice in leech Hillarin. This suggests that the leech Hillarin sequence originated from a duplication event of an ancestral protein with single cassette structure.


Assuntos
Axônios/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar , Humanos , Imuno-Histoquímica , Sanguessugas , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/genética , Homologia de Sequência de Aminoácidos
9.
Cell ; 105(4): 433-43, 2001 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-11371341

RESUMO

To analyze the function of the chromosomal kinase JIL-1, we generated an allelic series of hypomorphic and null mutations. JIL-1 is an essential kinase for viability, and reduced levels of JIL-1 kinase activity lead to a global change in chromatin structure. In JIL-1 hypomorphs, euchromatic regions of polytene chromosomes are severely reduced and the chromosome arms condensed. This is correlated with decreased levels of histone H3 Ser10 phosphorylation. These levels can be restored by a JIL-1 transgene placing JIL-1 directly in the pathway mediating histone H3 phosphorylation. We propose a model where JIL-1 kinase activity is required for maintaining the structure of the more open chromatin regions that facilitate gene transcription.


Assuntos
Drosophila/crescimento & desenvolvimento , Histonas/metabolismo , Proteínas de Insetos/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Cromatina Sexual/enzimologia , Alelos , Animais , Cromossomos/metabolismo , Drosophila/genética , Drosophila/metabolismo , Feminino , Testes Genéticos , Proteínas de Insetos/genética , Larva/fisiologia , Masculino , Mutação/fisiologia , Neurônios/fisiologia , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Cromossomos Sexuais/fisiologia , Fatores Sexuais , Transgenes/fisiologia
10.
Biochim Biophys Acta ; 1479(1-2): 1-14, 2000 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-11004526

RESUMO

Tractin is a novel member of the Ig-superfamily which has a highly unusual structure. It contains six Ig domains, four FNIII-like domains, an acidic domain, 12 repeats of a novel proline- and glycine-rich motif with sequence similarity to collagen, a transmembrane domain, and an intracellular tail with an ankyrin and a PDZ domain binding motif. By generating domain-specific antibodies, we show that Tractin is proteolytically processed at two cleavage sites, one located in the third FNIII domain, and a second located just proximal to the transmembrane domain resulting in the formation of four fragments. The most NH(2)-terminal fragment which is glycosylated with the Lan3-2, Lan4-2, and Laz2-369 glycoepitopes is secreted, and we present evidence which supports a model in which the remaining fragments combine to form a secreted homodimer as well as a transmembrane heterodimer. The extracellular domain of the dimers is mostly made up of the collagen-like PG/YG-repeat domain but also contains 11/2 FNIII domain and the acidic domain. The collagen-like PG/YG-repeat domain could be selectively digested by collagenase and we show by yeast two-hybrid analysis that the intracellular domain of Tractin can interact with ankyrin. Thus, the transmembrane heterodimer of Tractin constitutes a novel protein domain configuration where sequence that has properties similar to that of extracellular matrix molecules is directly linked to the cytoskeleton through interactions with ankyrin.


Assuntos
Axônios , Moléculas de Adesão Celular Neuronais/metabolismo , Imunoglobulinas/metabolismo , Processamento de Proteína Pós-Traducional , Animais , Anticorpos/imunologia , Moléculas de Adesão Celular Neuronais/imunologia , Moléculas de Adesão Celular Neuronais/fisiologia , Membrana Celular/metabolismo , Colagenases/metabolismo , Epitopos/metabolismo , Glicosilação , Sanguessugas
11.
J Cell Biol ; 149(5): 1005-10, 2000 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-10831604

RESUMO

JIL-1 is a novel chromosomal kinase that is upregulated almost twofold on the male X chromosome in Drosophila. Here we demonstrate that JIL-1 colocalizes and physically interacts with male specific lethal (MSL) dosage compensation complex proteins. Furthermore, ectopic expression of the MSL complex directed by MSL2 in females causes a concomitant upregulation of JIL-1 to the female X that is abolished in msl mutants unable to assemble the complex. Thus, these results strongly indicate JIL-1 associates with the MSL complex and further suggests JIL-1 functions in signal transduction pathways regulating chromatin structure.


Assuntos
Cromatina/enzimologia , Proteínas de Drosophila , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Cromossomo X/enzimologia , Animais , Proteínas de Ligação a DNA , Drosophila , Feminino , Dosagem de Genes , Regulação Enzimológica da Expressão Gênica/fisiologia , Histonas/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Masculino , Fosforilação , Ligação Proteica/genética , Transcrição Gênica
12.
Dev Biol ; 218(2): 137-45, 2000 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-10656758

RESUMO

Previous studies have indicated that the formation of stereotyped segmental nerves in leech embryos depends on the interactions between CNS projections and ingrowing afferents from peripheral neurons. Especially, CNS-ablation experiments have suggested that CNS-derived guidance cues are required for the correct navigation of several groups of peripheral sensory neurons. In order to directly test this hypothesis we have performed transplantations of CNS ganglia into ectopic sites in segments from which the resident ganglia have been removed. We find that the transplanted ganglia extend numerous axons distributed roughly equally in all directions. When these CNS projections reach and make contact with peripheral sensory axons they are used as guides for peripheral neurons to grow toward and into the ectopic ganglia even when this means following novel pathways that cross the midline and/or segmental boundaries. The peripheral sensory axons turn and grow toward the ectopic ganglia only when in physical contact with CNS axons, suggesting that diffusible chemoattractants are not a factor. These results demonstrate that the guidance cues provided by ectopic CNS projections are both necessary and sufficient to steer peripheral sensory neuron axons into the CNS.


Assuntos
Axônios , Sistema Nervoso Central/embriologia , Sanguessugas/embriologia , Neurônios Aferentes/citologia , Sistema Nervoso Periférico/embriologia , Animais , Sistema Nervoso Central/fisiologia , Sistema Nervoso Periférico/citologia
13.
J Cell Biol ; 151(7): 1401-12, 2000 Dec 25.
Artigo em Inglês | MEDLINE | ID: mdl-11134070

RESUMO

A spindle matrix has been proposed to help organize and stabilize the microtubule spindle during mitosis, though molecular evidence corroborating its existence has been elusive. In Drosophila, we have cloned and characterized a novel nuclear protein, skeletor, that we propose is part of a macromolecular complex forming such a spindle matrix. Skeletor antibody staining shows that skeletor is associated with the chromosomes at interphase, but redistributes into a true fusiform spindle structure at prophase, which precedes microtubule spindle formation. During metaphase, the spindle, defined by skeletor antibody labeling, and the microtubule spindles are coaligned. We find that the skeletor-defined spindle maintains its fusiform spindle structure from end to end across the metaphase plate during anaphase when the chromosomes segregate. Consequently, the properties of the skeletor-defined spindle make it an ideal substrate for providing structural support stabilizing microtubules and counterbalancing force production. Furthermore, skeletor metaphase spindles persist in the absence of microtubule spindles, strongly implying that the existence of the skeletor-defined spindle does not require polymerized microtubules. Thus, the identification and characterization of skeletor represents the first direct molecular evidence for the existence of a complete spindle matrix that forms within the nucleus before microtubule spindle formation.


Assuntos
Proteínas Cromossômicas não Histona/metabolismo , Proteínas de Drosophila , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Mitose , Proteínas Associadas à Matriz Nuclear , Fuso Acromático/química , Fuso Acromático/metabolismo , Transporte Ativo do Núcleo Celular , Sequência de Aminoácidos , Animais , Anticorpos/farmacologia , Sequência de Bases , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Proteínas Cromossômicas não Histona/antagonistas & inibidores , Proteínas Cromossômicas não Histona/química , Proteínas Cromossômicas não Histona/genética , Clonagem Molecular , Drosophila melanogaster/química , Drosophila melanogaster/embriologia , Imuno-Histoquímica , Dados de Sequência Molecular , Nocodazol/farmacologia , Fases de Leitura Aberta/genética , RNA Mensageiro/análise , RNA Mensageiro/genética , Alinhamento de Sequência , Fuso Acromático/efeitos dos fármacos , Fuso Acromático/genética , Temperatura
14.
Thromb Res ; 96(5): 355-63, 1999 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-10605950

RESUMO

The effect of red wine drinking was tested on fibrinolytic parameters and blood cells in nine healthy students at rest and after acute exercise. The subjects were randomly assigned in a crossover design to one of three treatment regimes: control situation, low-dose wine group, and high-dose wine group. Blood samples were drawn just prior to experimental start, at 2 and 4 hours, and the next morning at 8:00 a.m., at 8:30 a.m. just after exercise, and 2 hours after exercise. The fibrinolytic potential was measured by whole blood clot lysis time (WBCLT), tissue plasminogen activator, and plasminogen activator inhibitor-1 (PAI-1) antigens in plasma. A whole blood system was used to test the reactivity of blood cells by stimulating hirudinized blood with 5 ng/mL lipopolyusaccharide (LPS) for 2 hours at 37 degrees C and measurements of tissue necrosis factor alpha and interleukin-8 (IL-8) in the plasma. Intake of red wine caused impaired fibrinolysis shown by prolonged WBCLT (3.6, 20.7, and 55.7%, respectively, for control, low- and high-dose wine groups) due to increase in PAI-1 antigen (-0.8, 4.8, and 11.0 ng/mL, respectively, in the three groups). There was no effect of the red wine the next morning on the fibrinolytic system. A strong correlation was observed between WBCLT and PAI-1 antigen (p<0.0001). Acute exercise caused an immediate rise in both tissue plasminogen activator antigen and PAI-1 antigen levels and WBCLT was significantly shortened. In contrast to that of the wine groups, 2 hours after exercise WBCLT was prolonged in the control group, but not significantly so. Thus the red wine has a negative effect on the fibrinolytic system during rest, but may have a positive effect after strenuous exercise. The red wine had no immediate effect on LPS-induced tissue necrosis factor alpha or IL-8 production, although there was a tendency for higher cytokine production in the control group compared to the wine groups during and just after intake of wine. The next morning after exercise, the LPS-induced IL-8 production increased 137, 89, and 96%, respectively, in control, low-, and high-dose wine groups, probably due to a rise in epinephrine and activation of platelets. Although not significantly so, there was a tendency for red wine intake in the evening to suppress the reactivity of the cells after physical exercise the subsequent morning. It is suggested that the negative effect of red wine ingestion may be due to the toxic effect of ethanol on hepatocytes or adipose tissue and subsequent release of PAI-1, whereas the positive effect may be due to the red wine suppression of platelet activation and release of PAI-1 from activated platelets. It is proposed that at least part of the beneficial effect of red wine ingestion may be associated with the downregulation of cytokine production.


Assuntos
Exercício Físico , Fibrinólise/efeitos dos fármacos , Vinho/efeitos adversos , Adulto , Antígenos , Testes de Coagulação Sanguínea , Estudos Cross-Over , Relação Dose-Resposta a Droga , Humanos , Interleucina-8/sangue , Lipopolissacarídeos/farmacologia , Masculino , Periodicidade , Inibidor 1 de Ativador de Plasminogênio/sangue , Inibidor 1 de Ativador de Plasminogênio/imunologia , Descanso , Fatores de Tempo , Ativador de Plasminogênio Tecidual/sangue , Ativador de Plasminogênio Tecidual/imunologia , Fator de Necrose Tumoral alfa/metabolismo
15.
Biochim Biophys Acta ; 1452(2): 161-71, 1999 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-10559469

RESUMO

LeechCAM is a recently described member of the Ig-superfamily which has five Ig-domains, two FNIII-domains, a transmembrane domain, and a cytoplasmic domain. Phylogenetic analysis indicated that LeechCAM is the leech homolog of apCAM, FasII, and vertebrate NCAM. Using a leechCAM-specific monoclonal antibody we show by immunoblot analysis and by Triton X-114 phase separation experiments that in addition to existing in a transmembrane version LeechCAM is likely to be proteolytically cleaved into a secreted form without the transmembrane domain and the intracellular tail. Furthermore, by immunoprecipitation we demonstrate that LeechCAM is glycosylated with the Laz2-369 glycoepitope, an epitope that has been specifically implicated in regulation of axonal outgrowth and synapse formation.


Assuntos
Moléculas de Adesão Celular/metabolismo , Sanguessugas/metabolismo , Animais , Moléculas de Adesão Celular/química , Endopeptidases , Proteína Ligante Fas , Glicosilação , Sanguessugas/embriologia , Glicoproteínas de Membrana/metabolismo , Neurônios/metabolismo , Octoxinol , Filogenia , Polietilenoglicóis , Testes de Precipitina , Isoformas de Proteínas/metabolismo
16.
Mol Cell ; 4(1): 129-35, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10445035

RESUMO

We have cloned and characterized JIL-1, a novel tandem kinase in Drosophila that associates with the chromosomes throughout the cell cycle. Antibody staining and live imaging of JIL-1-GFP transgenic flies show that JIL-1 localizes to the gene-rich interband regions of larval polytene chromosomes and is upregulated almost 2-fold on the hypertranscribed male X chromosome compared to autosomes. Phylogenetic analysis suggests that JIL-1 together with human MSKs defines a separate family of tandem kinases. That JIL-1 is a functional kinase was demonstrated by autophosphorylation and phosphorylation of histone H3 in vitro. Based on these findings, we propose that JIL-1 may play a role in transcriptional control potentially by regulating chromatin structure.


Assuntos
Proteínas Cromossômicas não Histona/genética , Drosophila/enzimologia , Proteínas Quinases/genética , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Ciclo Celular , Cromatina/química , Proteínas Cromossômicas não Histona/química , Clonagem Molecular , Drosophila/embriologia , Regulação da Expressão Gênica , Histonas/metabolismo , Humanos , Proteínas de Insetos/química , Proteínas de Insetos/genética , Masculino , Dados de Sequência Molecular , Filogenia , Proteínas Quinases/química , Glândulas Salivares/enzimologia , Cromossomo X/genética
17.
J Neurobiol ; 40(2): 244-53, 1999 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10413454

RESUMO

Using monoclonal antibodies, we have identified two novel intermediate filament (IF) proteins, Gliarin and Macrolin, which are specifically expressed in the central nervous system of an invertebrate. The two proteins both contain the coiled-coil rod domain typical of the superfamily of IF proteins flanked by unique N- and C-terminal domains. Gliarin was found in all glial cells including macro- and microglial cells, whereas Macrolin was expressed in only a single pair of giant connective glial cells. The identification of Macrolin and Gliarin together with the characterization of the strictly neuronal IF protein Filarin in leech central nervous system demonstrate that multiple neuron- and glial-specific IFs are not unique to the vertebrate nervous system but are also found in invertebrates. Interestingly, phylogenetic analysis based on maximum parsimony indicated that the presence of neuron- and glial cell-specific IFs in coelomate protostomes as well as in vertebrates is not of monophyletic origin, but rather represents convergent evolution and appears to have arisen independently.


Assuntos
Sistema Nervoso Central/metabolismo , Proteínas de Filamentos Intermediários/isolamento & purificação , Sanguessugas/metabolismo , Proteínas do Tecido Nervoso/isolamento & purificação , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , DNA Complementar/genética , Expressão Gênica , Proteínas de Filamentos Intermediários/biossíntese , Proteínas de Filamentos Intermediários/genética , Proteínas de Filamentos Intermediários/imunologia , Sanguessugas/genética , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/imunologia , Filogenia , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos
18.
Biochim Biophys Acta ; 1449(2): 194-8, 1999 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-10082978

RESUMO

Using a polyclonal antiserum a cDNA encoding a Rab escort protein (REP) homolog in Drosophila has been identified and sequenced. The gene encodes a 511 residue protein with a predicted molecular mass of 56855 Da. Antibody labeling demonstrates that Drosophila REP protein is present in the early embryo and that it is being apportioned uniformly throughout the embryo in a process likely to be linked to the syncytial nuclear divisions. In situ hybridization to polytene chromosomes reveals that the Drosophila REP gene is located in the 56E region on the second chromosome. Drosophila REP is the first invertebrate REP homolog to be identified and characterized.


Assuntos
Alquil e Aril Transferases , Proteínas de Transporte/química , Sequência Conservada , Drosophila/metabolismo , Proteínas rab de Ligação ao GTP , Sequência de Aminoácidos , Animais , Anticorpos/imunologia , Proteínas de Transporte/genética , Proteínas de Transporte/imunologia , Mapeamento Cromossômico , Drosophila/embriologia , Drosophila/genética , Dados de Sequência Molecular , Alinhamento de Sequência
19.
J Neurobiol ; 38(2): 287-99, 1999 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-10022573

RESUMO

We have cloned a novel voltage-gated K channel, LKv1, in two species of leech. The properties of LKv1 expressed in transiently transfected HEK293 cells is that of a delayed rectifier current. LKv1 may be a major modulator of excitability in leech neurons, since antibody localization studies show that LKv1 is expressed in the soma and axons of all neurons in both the central and peripheral nervous systems. Comparison of the biophysical and pharmacological properties of LKv1 with native voltage-gated conductances in leech neurons suggests that LKv1 may correspond to the previously characterized delayed rectifier current, I(K). Phylogenetic analysis of LKv1 shows that it is related to the Shaker subfamily of voltage-gated K channels although it occupies a separate branch from that of the monophyletic Shaker clade composed of the flatworm, Aplysia, Drosophila, and mammalian Shaker homologs as well as from that of two recently identified Shaker-related K channels in jellyfish. Thus, this analysis indicates that this group of voltage-gated K channels contains several evolutionarily divergent lineages.


Assuntos
Sanguessugas/fisiologia , Neurônios/fisiologia , Canais de Potássio/biossíntese , Sequência de Aminoácidos , Animais , Evolução Biológica , Western Blotting , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Eletrofisiologia , Imuno-Histoquímica , Ativação do Canal Iônico/fisiologia , Potenciais da Membrana/fisiologia , Dados de Sequência Molecular , Técnicas de Patch-Clamp , Filogenia , Canais de Potássio/genética , Superfamília Shaker de Canais de Potássio
20.
Crit Rev Eukaryot Gene Expr ; 9(3-4): 267-77, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10651243

RESUMO

Nuclear architecture is remodeled during interphase in response to changes in gene activity as well as to changing structural and functional requirements during cell division. Using the monoclonal antibody mAb2A, we have identified two proteins that appear to play important roles in these processes: JIL-1 is a tandem serine-threonine kinase implicated in the regulation of chromatin structure, whereas Skeletor is a novel protein participating in structural nuclear remodeling during the cell cycle. Antibody staining and live imaging of JIL-1-GFP transgenic flies show that JIL-1 localizes to the gene-rich interband regions of larval polytene chromosomes and is upregulated almost twofold on the hypertranscribed male X chromosome compared with autosomes. We propose that JIL-1 may play a role in transcriptional control potentially by regulating chromatin structure. The other mAb2A antigen, Skeletor, is distributed in a nuclear meshwork pattern that can be observed in stereo pair images to reorganize during the cell cycle to form a spindle-like structure at prometaphase that is distinct from the microtubule spindle apparatus. Taking advantage of the powerful molecular and genetic approaches offered in Drosophila, the study of these two proteins promises to yield new insight into what defines nuclear architecture at the molecular level and how its remodeling is regulated.


Assuntos
Núcleo Celular/genética , Cromatina/genética , Sequência de Aminoácidos , Animais , Núcleo Celular/química , Núcleo Celular/ultraestrutura , Cromatina/química , Cromatina/ultraestrutura , Drosophila , Feminino , Regulação da Expressão Gênica , Humanos , Masculino , Dados de Sequência Molecular , Proteínas Serina-Treonina Quinases , Alinhamento de Sequência
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