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1.
Cereb Cortex ; 31(9): 4357-4375, 2021 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-33914862

RESUMO

The frontal cortex-basal ganglia network plays a pivotal role in adaptive goal-directed behaviors. Medial frontal cortex (MFC) encodes information about choices and outcomes into sequential activation of neural population, or neural trajectory. While MFC projects to the dorsal striatum (DS), whether DS also displays temporally coordinated activity remains unknown. We studied this question by simultaneously recording neural ensembles in the MFC and DS of rodents performing an outcome-based alternative choice task. We found that the two regions exhibited highly parallel evolution of neural trajectories, transforming choice information into outcome-related information. When the two trajectories were highly correlated, spike synchrony was task-dependently modulated in some MFC-DS neuron pairs. Our results suggest that neural trajectories concomitantly process decision-relevant information in MFC and DS with increased spike synchrony between these regions.


Assuntos
Comportamento de Escolha/fisiologia , Corpo Estriado/fisiologia , Córtex Pré-Frontal/fisiologia , Desempenho Psicomotor/fisiologia , Animais , Masculino , Ratos , Ratos Long-Evans
2.
Eur J Neurosci ; 48(10): 3171-3185, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30118167

RESUMO

Human and non-human primates can readily perceive the shape of objects using visual motion. Classically, shape, and motion are considered to be separately processed via ventral and dorsal cortical pathways, respectively. However, many lines of anatomical and physiological evidence have indicated that these two pathways are likely to be interconnected at some stage. For motion-defined shape perception, these two pathways should interact with each other because the ventral pathway must utilize motion, which the dorsal pathway processes, to extract shape signal. However, it is unknown how interactions between cortical pathways are involved in neural mechanisms underlying motion-defined shape perception. We review evidence from psychophysical, lesion, neuroimaging and physiological research on motion-defined shape perception and then discuss the effects of behavioral demands on neural activity in ventral and dorsal cortical areas. Further, we discuss functions of two candidate sets of levels: early and higher-order cortical areas. The extrastriate area V4 and middle temporal (MT) area, which are reciprocally connected, at the early level are plausible areas for extracting the shape and/or constituent parts of shape from motion cues because neural dynamics are different from those during luminance-defined shape perception. On the other hand, among other higher-order visual areas, the anterior superior temporal sulcus likely contributes to the processing of cue-invariant shape recognition rather than cue-dependent shape processing. We suggest that sharing information about motion and shape between the early visual areas in the dorsal and ventral pathways is dependent on visual cues and behavioral requirements, indicating the interplay between the pathways.


Assuntos
Córtex Cerebral/fisiologia , Percepção de Forma/fisiologia , Percepção de Movimento/fisiologia , Primatas/fisiologia , Vias Visuais/fisiologia , Animais , Humanos
3.
Cereb Cortex ; 27(7): 3818-3831, 2017 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-28184411

RESUMO

Neurons in medial frontal cortex (MFC) receive sensory signals that are crucial for decision-making behavior. While decision-making is easy for familiar sensory signals, it becomes more elaborative when sensory signals are less familiar to animals. It remains unclear how the population of neurons enables the coordinate transformation of such a sensory input into ambiguous choice responses. Furthermore, whether and how cortical oscillations temporally coordinate neuronal firing during this transformation has not been extensively studied. Here, we recorded neuronal population responses to familiar or unfamiliar auditory cues in rat MFC and computed their probabilistic evolution. Population responses to familiar sounds organize into neuronal trajectories containing multiplexed sensory, motor, and choice information. Unfamiliar sounds, in contrast, evoke trajectories that travel under the guidance of familiar paths and eventually diverge to unique decision states. Local field potentials exhibited beta- (15-20 Hz) and gamma-band (50-60 Hz) oscillations to which neuronal firing showed modest phase locking. Interestingly, gamma oscillation, but not beta oscillation, increased its power abruptly at some timepoint by which neural trajectories for different choices were near maximally separated. Our results emphasize the importance of the evolution of neural trajectories in rapid probabilistic decisions that utilize unfamiliar sensory information.


Assuntos
Vias Aferentes/fisiologia , Mapeamento Encefálico , Comportamento de Escolha/fisiologia , Lobo Frontal/fisiologia , Sensação/fisiologia , Potenciais de Ação/fisiologia , Animais , Sinais (Psicologia) , Estimulação Elétrica , Lobo Frontal/citologia , Lateralidade Funcional/fisiologia , Masculino , Rede Nervosa/fisiologia , Neurônios/fisiologia , Análise de Componente Principal , Ratos , Ratos Long-Evans
4.
J Stroke Cerebrovasc Dis ; 27(7): 2032-2034, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29598906

RESUMO

BACKGROUND AND PURPOSE: The posterior meningeal artery (PMA) is known as a dura mater-nourishing vessel. We encountered a patient with Wallenberg syndrome during transarterial embolization of the PMA associated with the dural arteriovenous fistula (DAVF). METHODS: After development of Wallenberg syndrome in the patient, we assessed origins of the PMA patterns in 300 cases and divided them into 3 types. CASE PRESENTATION: A 63-year-old man was incidentally diagnosed as having transverse-sigmoid sinus DAVF with a cortical venous reflux. During the transarterial embolization, the patient complained of vertigo and numbness of the right extremities. Postoperatively, the patient exhibited Wallenberg syndrome. Diffusion-weighted magnetic resonance imaging showed a high-intensity area on the lateral side of the right medulla. CONCLUSIONS: While performing arterial embolization of the PMA that directly originates from the intracranial vertebral artery, the possibility of deficient brainstem nourishment must be considered.


Assuntos
Síndrome Medular Lateral/patologia , Artérias Meníngeas/anormalidades , Artérias Meníngeas/anatomia & histologia , Variação Biológica Individual , Diagnóstico Diferencial , Humanos , Síndrome Medular Lateral/diagnóstico , Síndrome Medular Lateral/diagnóstico por imagem , Síndrome Medular Lateral/cirurgia , Masculino , Artérias Meníngeas/diagnóstico por imagem , Artérias Meníngeas/cirurgia , Pessoa de Meia-Idade , Artéria Vertebral/anormalidades , Artéria Vertebral/diagnóstico por imagem , Artéria Vertebral/cirurgia
5.
J Phys Ther Sci ; 29(1): 8-11, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28210028

RESUMO

[Purpose] The mechanical strength of wheelchair and seating products is specified by standards to ensure user safety. However, some individual users are affected by severe spasticity resulting from cerebral palsy that occasionally breaks such devices. Until recently, reports of quantitative measurement of these mechanical overloads have been scarce, and the exact loads applied have been unknown. This study aimed to conduct a sampling survey prior to conducting a future full-scale study. [Subjects and Methods] Using two force plates, we measured the load on the foot support applied by three participants with cerebral palsy. The applied load was measured at a 100 Hz sampling rate, and maximum load and vertical/horizontal component forces were analyzed. [Results] The maximum load applied by participants was approximately 1.34 times their body weight. The vertical force applied on the foot support accounted for 70-80% of the total load. The horizontal force also reached about 35% at its maximum. [Conclusion] Because the horizontal load also reached 1/3 of the total load, it will be advantageous to use a three-axis force plate to measure forces in a future study with more participants.

6.
J Neurosci ; 33(25): 10209-20, 2013 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-23785137

RESUMO

It is widely accepted that dorsal striatum neurons participate in either the direct pathway (expressing dopamine D1 receptors) or the indirect pathway (expressing D2 receptors), controlling voluntary movements in an antagonistically balancing manner. The D1- and D2-expressing neurons are activated and inactivated, respectively, by dopamine released from substantia nigra neurons encoding reward expectation. However, little is known about the functional representation of motor information and its reward modulation in individual striatal neurons constituting the two pathways. In this study, we juxtacellularly recorded the spike activity of single neurons in the dorsolateral striatum of rats performing voluntary forelimb movement in a reward-predictable condition. Some of these neurons were identified morphologically by a combination of juxtacellular visualization and in situ hybridization for D1 mRNA. We found that the striatal neurons exhibited distinct functional activations before and during the forelimb movement, regardless of the expression of D1 mRNA. They were often positively, but rarely negatively, modulated by expecting a reward for the correct motor response. The positive reward modulation was independent of behavioral differences in motor performance. In contrast, regular-spiking and fast-spiking neurons in any layers of the motor cortex displayed only minor and unbiased reward modulation of their functional activation in relation to the execution of forelimb movement. Our results suggest that the direct and indirect pathway neurons cooperatively rather than antagonistically contribute to spatiotemporal control of voluntary movements, and that motor information is subcortically integrated with reward information through dopaminergic and other signals in the skeletomotor loop of the basal ganglia.


Assuntos
Movimento/fisiologia , Neostriado/fisiologia , Neurônios/fisiologia , Recompensa , Animais , Gânglios da Base/fisiologia , Biotina/análogos & derivados , Condicionamento Operante/fisiologia , Dextranos , Fenômenos Eletrofisiológicos , Corantes Fluorescentes , Membro Anterior/fisiologia , Hibridização In Situ , Masculino , Córtex Motor/citologia , Córtex Motor/fisiologia , Neostriado/citologia , Rede Nervosa/fisiologia , Ratos , Ratos Long-Evans , Receptores de Dopamina D1/genética , Receptores de Dopamina D1/fisiologia , Receptores de Dopamina D2/genética , Receptores de Dopamina D2/fisiologia
7.
Biol Pharm Bull ; 34(5): 779-82, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21532173

RESUMO

Stems of genus Dendrobium (Orchidaceae) have been traditionally used as an herbal medicine (Dendrobii Herba) in Eastern Asia. Although demand for Dendrobium is increasing rapidly, wild resources are decreasing due to over-collection. This study aimed to identify plant sources of Dendrobii Herba on the market based on sequences of the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA. We constructed an ITS1-5.8S-ITS2 sequence database of 196 Dendrobium species, and the database was employed to identify 21 herbal samples. We found that 13 Dendrobium species (D. catenatum, D. cucullatum, D. denudans, D. devonianum, D. eriiflorum, D. hancockii, D. linawianum, D. lituiflorum, D. loddigesii, D. polyanthum, D. primulinum, D. regium, and D. transparens) were possibly used as plant sources of Dendrobii Herba, and unidentified species allied to D. denudans, D. eriiflorum, D. gregulus, or D. hemimelanoglossum were also used as sources. Furthermore, it is clear that D. catenatum is one of the most important sources of Dendrobii Herba (5 out of 21 samples).


Assuntos
DNA Ribossômico/efeitos dos fármacos , Dendrobium/química , Medicina Herbária , Sequência de Bases , Primers do DNA , Reação em Cadeia da Polimerase
8.
BMC Psychiatry ; 9: 38, 2009 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-19531229

RESUMO

BACKGROUND: Adherence has recently been suggested to be divided into these two components: persistence (i.e., whether patients continue treatment or not) and compliance (i.e., whether patients take doses as instructed). However, no study has yet assessed these two clinically relevant components at the same time in adherence to antidepressant treatment in the clinical outpatient setting. METHODS: In this retrospective chart-review, 6-month adherence to antidepressants was examined in 367 outpatients with a major depressive disorder (ICD-10) (170 males; mean +/- SD age 37.6 +/- 13.9 years), who started antidepressant treatment from April 2006 through March 2007. Additionally, we evaluated Medication Possession Rate (MPR), defined as the total days a medication was dispensed to patients divided by the treatment period. RESULTS: Only 161 patients (44.3%) continued antidepressant treatment for 6 months. Among 252 patients who discontinued their initial antidepressant, 63.1% of these patients did so without consulting their physicians. Sertraline use was associated with a higher persistence rate at month 6 (odds ratio 2.59 in comparison with sulpiride), and the use of anxiolytic benzodiazepines had a positive effect on persistence to antidepressant treatment only at month 1 (odds ratio 2.14). An overall MPR was 0.77; 55.6% of patients were considered compliant (i.e., a MPR of > or = 0.8). CONCLUSION: Given a high rate of antidepressant discontinuation without consulting their physicians, closer communication between patients and their physicians should be encouraged. Although the use of anxiolytic benzodiazepines was associated with a higher persistence to antidepressant treatment at month 1, the use of these drugs should be avoided as a rule, given their well-known serious adverse effects.


Assuntos
Antidepressivos/uso terapêutico , Transtorno Depressivo Maior/tratamento farmacológico , Adesão à Medicação/estatística & dados numéricos , Cooperação do Paciente/estatística & dados numéricos , Adolescente , Idoso , Idoso de 80 Anos ou mais , Antidepressivos/administração & dosagem , Benzodiazepinas/uso terapêutico , Transtorno Depressivo Maior/psicologia , Feminino , Humanos , Japão , Masculino , Adesão à Medicação/psicologia , Pessoa de Meia-Idade , Cooperação do Paciente/psicologia , Pacientes Desistentes do Tratamento , Relações Médico-Paciente , Estudos Retrospectivos , Sertralina/uso terapêutico
9.
Appl Microbiol Biotechnol ; 80(6): 1007-13, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18633609

RESUMO

An alpha-L-rhamnosidase was purified by fractionating a culture filtrate of Aspergillus kawachii grown on L-rhamnose as the sole carbon source. The alpha-L-rhamnosidase had a molecular mass of 90 kDa and a high degree of N-glycosylation of approximately 22%. The enzyme exhibited optimal activity at pH 4.0 and temperature of 50 degrees C. Further, it was observed to be thermostable, and it retained more than 80% of its original activity following incubation at 60 degrees C for 1 h. Its T (50) value was determined to be 72 degrees C. The enzyme was able to hydrolyze alpha-1,2- and alpha-1,6-glycosidic bonds. The specific activity of the enzyme was higher toward naringin than toward hesperidin. The A. kawachii alpha-L-rhamnosidase-encoding gene (Ak-rhaA) codes for a 655-amino-acid protein. Based on the amino acid sequence deduced from the cDNA, the protein possessed 13 potential N-glycosylation recognition sites and exhibited a high degree of sequence identity (up to 75%) with the alpha-L-rhamnosidases belonging to the glycoside hydrolase family 78 from Aspergillus aculeatus and with hypothetical Aspergillus oryzae and Aspergillus fumigatus proteins.


Assuntos
Aspergillus/enzimologia , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Aspergillus/genética , DNA Fúngico/química , DNA Fúngico/genética , Estabilidade Enzimática , Flavanonas/metabolismo , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/isolamento & purificação , Glicosilação , Hesperidina/metabolismo , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Peso Molecular , Ramnose/metabolismo , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , Temperatura , Temperatura de Transição
10.
J Plant Physiol ; 228: 85-91, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29879604

RESUMO

Flower color patterns are the result of spatially and temporally restricted pigment deposition, and clarifying the mechanisms responsible for restricted pigment deposition is a topic of broad interest for both theoretical and practical reasons. The Oriental hybrid lily cultivar 'Dizzy' develops red stripes along the tepal midribs; in order to clarify the genetic basis of these stripes, we isolated most of the genes related to anthocyanin accumulation from 'Dizzy' tepals and compared their expression levels between the red stripe region and the white marginal region of the tepals. RNA-seq revealed a complete set of genes necessary for anthocyanin biosynthesis and transport, including anthocyanidin 3-O-glucosyltransferase and glutathione S-transferase. Most of these genes were expressed at higher rates in the red stripe region than in the white region, suggesting that transcriptional regulation of these genes is primarily responsible for the spatially restricted anthocyanin deposition in 'Dizzy' tepals. Subgroup 6 R2R3-MYB is a major factor regulating anthocyanin biosynthesis: RNA-seq clarified three subgroup 6 R2R3-MYB genes expressed in 'Dizzy' tepals, of which MYB12 was predominantly expressed. Expression of MYB12 was six-fold higher in the red-pigmented region than in the white region. Thus, MYB12 is more likely to be involved in the regulation of the restricted anthocyanin deposition in 'Dizzy', even though MYB12 is expressed in the entire tepal region of many Oriental hybrid lily cultivars. Diversity of the expression profiles of MYB12 among lily cultivars and species is also discussed.


Assuntos
Antocianinas/metabolismo , Flores/metabolismo , Flores/fisiologia , Lilium/metabolismo , Lilium/fisiologia , Proteínas de Plantas/metabolismo , Antocianinas/genética , Flores/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Lilium/genética , Pigmentação/genética , Pigmentação/fisiologia , Proteínas de Plantas/genética
11.
J Neurosurg ; 131(1): 104-108, 2018 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-29979114

RESUMO

Intraventricular hemorrhage and intracerebral aneurysms are relatively frequent complications associated with moyamoya disease. Prevention of aneurysm rerupture is important because it significantly decreases the morbidity and mortality rates. Aneurysms arising distal to collateral flow are sometimes observed in patients with intraventricular hemorrhage; however, the treatment of these aneurysms remains challenging because of their deep-seated location in the brain and accompanying narrow surgical corridor. The authors describe a neuroendoscopic aneurysm clipping technique performed in 2 cases using a small-diameter tubular retractor for intraventricular aneurysms of the distal lateral posterior choroidal artery. In this technique, the surgical field was continuously irrigated with artificial CSF to keep the ventricle size intact, and aneurysm clipping was performed through a tubular retractor that was introduced with neuronavigational guidance. The patients' postoperative courses were uneventful, and CT angiography revealed complete clipping of the aneurysms and patent parent arteries. Endoscopic clipping using a tubular retractor is an effective and less invasive alternative for treating intraventricular aneurysms. The wet-field endoscopic technique is performed in an aqueous field and maintains an intact ventricle size, allowing for a clear surgical view and a wider, enhanced surgical field.

12.
Nat Neurosci ; 21(12): 1764-1773, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30420732

RESUMO

In the brain, decision making is instantiated in dedicated neural circuits. However, there is considerable individual variability in decision-making behavior, particularly under uncertainty. The origins of decision variability within these conserved neural circuits are not known. Here we demonstrate in the rat medial frontal cortex (MFC) that individual variability is a consequence of altered stability in neuronal populations. In a sensory-guided choice task, rats trained on familiar stimuli were exposed to unfamiliar stimuli, resulting in variable choice responses across individuals. We created a recurrent network model to examine the source of variability in MFC neurons, and found that the landscape of neural population trajectories explained choice variability across different unfamiliar stimuli. We experimentally confirmed model predictions showing that trial-by-trial variability in neuronal activity indexes the landscape and predicts individual variation. These results show that neural stability is a critical component of the MFC neural dynamics that underpins individual variation in decision-making.


Assuntos
Tomada de Decisões/fisiologia , Lobo Frontal/fisiologia , Rede Nervosa/fisiologia , Animais , Comportamento Animal/fisiologia , Comportamento de Escolha/fisiologia , Sinais (Psicologia) , Masculino , Modelos Neurológicos , Neurônios/fisiologia , Ratos
13.
Neuroreport ; 28(13): 793-799, 2017 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-28678113

RESUMO

In the primate brain, shape and motion are considered to be separately processed in the ventral and dorsal visual cortical areas, respectively. However, to achieve shape perception with a motion cue, shape and motion cannot be processed exclusively in separate cortical areas. Interactions between ventral and dorsal cortical areas are required, and yet, the neural mechanisms underlying motion-defined shape perception remain unclear. Here, we assessed the temporal properties of single-unit activity recorded from V4, the middle temporal area, and the anterior superior temporal sulcus while monkeys discriminated shapes defined by motion and luminance cues. Visual response latencies of V4 neurons were shorter in the luminance-cue condition than in the motion-cue condition. Meanwhile, the timings of initiation of shape selectivity were not different between cue conditions, indicating a difference in processing time. Middle temporal neurons were less shape modulated in the luminance-cue condition than in the motion-cue condition. Temporal properties of neural activities in the lower bank of anterior superior temporal sulcus were similar between cue conditions. These results suggest that an interaction of the ventral cortex with the dorsal cortex is required for shape discrimination with different visual cues.


Assuntos
Potenciais de Ação/fisiologia , Córtex Cerebral/citologia , Percepção de Forma/fisiologia , Percepção de Movimento/fisiologia , Neurônios/fisiologia , Animais , Córtex Cerebral/fisiologia , Macaca mulatta , Masculino , Estimulação Luminosa
14.
AoB Plants ; 62014 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-25107672

RESUMO

It is always difficult to construct coherent classification systems for plant lineages having diverse morphological characters. The genus Dendrobium, one of the largest genera in the Orchidaceae, includes ∼1100 species, and enormous morphological diversification has hindered the establishment of consistent classification systems covering all major groups of this genus. Given the particular importance of species in Dendrobium section Dendrobium and allied groups as floriculture and crude drug genetic resources, there is an urgent need to establish a stable classification system. To clarify phylogenetic relationships in Dendrobium section Dendrobium and allied groups, we analysed the macromolecular characters of the group. Phylogenetic analyses of 210 taxa of Dendrobium were conducted on DNA sequences of internal transcribed spacer (ITS) regions of 18S-26S nuclear ribosomal DNA and the maturase-coding gene (matK) located in an intron of the plastid gene trnK using maximum parsimony and Bayesian methods. The parsimony and Bayesian analyses revealed 13 distinct clades in the group comprising section Dendrobium and its allied groups. Results also showed paraphyly or polyphyly of sections Amblyanthus, Aporum, Breviflores, Calcarifera, Crumenata, Dendrobium, Densiflora, Distichophyllae, Dolichocentrum, Holochrysa, Oxyglossum and Pedilonum. On the other hand, the monophyly of section Stachyobium was well supported. It was found that many of the morphological characters that have been believed to reflect phylogenetic relationships are, in fact, the result of convergence. As such, many of the sections that have been recognized up to this point were found to not be monophyletic, so recircumscription of sections is required.

15.
Neuroreport ; 21(7): 532-6, 2010 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-20386346

RESUMO

To investigate the neuronal mechanisms of motion-defined shape processing, we examined single-neuron activity in area V4 in monkeys performing a shape discrimination task under the shape-from-motion (SFM) condition, in which a motion cue is critical for shape perception, and under the shape-from-luminance (SFL) condition, in which a luminance cue is critical. About 35% (30 of 86) of neurons responded selectively to shapes under both the SFM and SFL conditions. These neurons showed a similar shape preference between the SFM and SFL conditions. There was a weak tendency of lower shape selectivity in error trials. These results suggest the involvement of V4 in SFM processing as well as in SFL processing.


Assuntos
Discriminação Psicológica , Percepção de Forma/fisiologia , Percepção de Movimento/fisiologia , Neurônios/fisiologia , Córtex Visual/fisiologia , Animais , Mapeamento Encefálico , Processamento de Imagem Assistida por Computador , Macaca mulatta , Imageamento por Ressonância Magnética , Estimulação Luminosa
16.
Methods Mol Biol ; 589: 349-64, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20099114

RESUMO

Plants belonging to the Rhododendron subgenera Pentanthera (deciduous) and Tsutsusi and Azaleastrum (evergreen) are called azaleas. Concerning their mutual phylogenetic positions, the Pentanthera subgenus is closer to evergreen rhododendrons (subgenera Rhododendron and Hymenanthes) than to the Tsutsusi subgenus. Both azalea types are important ornamentals with a long breeding tradition. Different hybrid groups are often named after the supposed principal ancestor species. Molecular techniques for phylogenetic and kinship research have been evaluated to a great extent. First, some studies using comparative gene sequencing are presented; this approach was then widened to the use of molecular markers to reveal more detailed genetic relationships. Finally, the use of candidate genes as functional markers for the assessment of genetic diversity is presented. This opens new research lines to the genetic mapping of plant traits and azalea genomics.


Assuntos
Regulação da Expressão Gênica de Plantas , Genes de Plantas , Técnicas Genéticas , Filogenia , Rhododendron/genética , Evolução Molecular , Marcadores Genéticos , Variação Genética , Rhododendron/classificação , Análise de Sequência de DNA
17.
Biosci Biotechnol Biochem ; 70(10): 2476-80, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17031032

RESUMO

A unique N-linked glycosylation motif (Asn(79)-Tyr-Thr) was found in the sequence of type-A feruloyl esterases from Aspergillus spp. To clarify the function of the flap, the role of N-linked oligosaccharides located in the flap region on the biochemical properties of feruloyl esterase (AwFAEA) from Aspergillus awamori expressed in Pichia pastoris was analyzed by removing the N-linked glycosylation recognition site by site-directed mutagenesis. N79 was replaced with A or Q. N-glycosylation-free N79A and N79Q mutant enzymes had lower activity than that of the glycosylated recombinant AwFAEA wild-type enzyme toward alpha-naphthylbutyrate (C4), alpha-naphthylcaprylate (C8), and phenolic acid methyl esters. Kinetic analysis of the mutant enzymes indicated that the lower catalytic efficiency was due to a combination of increased Km and decreased k(cat) for N79A, and to a considerably decreased k(cat) for N79Q. N79A and N79Q mutant enzymes also exhibited considerably reduced thermostability relative to the wild-type.


Assuntos
Aspergillus/enzimologia , Hidrolases de Éster Carboxílico/química , Oligossacarídeos , Motivos de Aminoácidos , Substituição de Aminoácidos , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , Catálise , Estabilidade Enzimática , Ésteres/metabolismo , Glicosilação , Cinética , Mutagênese Sítio-Dirigida , Temperatura
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