• BMB Reports
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• 제47권9호
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• pp.518-523
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• 2014

The maternal transcription factor Dorsal (Dl) functions as both an activator and a repressor in a context-dependent manner to control dorsal-ventral patterning in the Drosophila embryo. Previous studies have suggested that Dl is an intrinsic activator and its repressive activity requires additional corepressors that bind corepressor-binding sites near Dl-binding sites. However, the molecular identities of the corepressors have yet to be identified. Here, we present evidence that Capicua (Cic) is involved in Dl-mediated repression in the zerkn$\ddot{u}$llt (zen) ventral repression element (VRE). Computational and genetic analyses indicate that a DNA-binding consensus sequence of Cic is highly analogous with previously identified corepressor-binding sequences and that Dl failed to repress zen expression in lateral regions of cic mutant embryos. Furthermore, electrophoretic mobility shift assay (EMSA) shows that Cic directly interacts with several corepressor-binding sites in the zen VRE. These results suggest that Cic may function as a corepressor by binding the VRE.

• Molecules and Cells
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• 제21권2호
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• pp.261-268
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• 2006

The Drosophila methuselah (mth) mutant has an approximately 35 percent increase in average lifespan, and enhanced resistance to various forms of stress, including starvation, high temperature, and dietary paraquat. To examine the transcriptional regulation of mth, we used luciferase assays employing Drosophila S2 cells. Two positive control elements were found at -542 ~ -272 (PE1) and +28 ~ +217 (PE2), where putative binding sites for transcription factors including Dorsal (Dl) were identified. Cotransfection of a Dl expression plasmid with a mth-luciferase reporter plasmid resulted in decreased reporter activity. PE1 and PE2, the minimal elements for strong promoter activity, were required for maximal repression by Dl protein. The N-terminal Rel homology domain (RHD) of Dl was not sufficient for repression of mth. We demonstrated by chromatin affinity precipitation (ChAP) assays in S2 cells that Dl bound to the putative PE1 binding site. Unexpectedly, semi-quantitative RT-PCR analysis revealed that the level of mth transcripts was reduced in dl flies. However, the in vivo result support the view that mth expression is regulated by dl, since it is well known that Dl functions as both a transcriptional activator and repressor depending on what other transcription factors are present. These findings suggest that both innate immunity and resistance to stress are controlled by Dl protein.

• 대한수의학회지
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• 제39권5호
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• pp.1013-1016
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• 1999

A four-year-old female Jindo dog was admitted to the hospital because of bilaterally symmetrical alopecia and inappentence. Systemic abnomalities detected on physical examination were obesity, bradycardia, exercise intolerance, mental dullness and hypothermia. Dermatologically symmetrical alopecia on the back, trunk and dorsal tail, and ventral hyperpigmentation were observed. Also face was very tragic. The level of basal $T_4$ was 0.01ug/dl. In TSH stimulation test $fT_4$ value was 0.08ng/dl. After treatment was initiated with L-thyroxine the dog's general condition was improved at 4 weeks.

• BMB Reports
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• 제49권10호
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• pp.572-577
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• 2016

The short gastrulation (sog) shadow enhancer directs early and late sog expression in the neurogenic ectoderm and the ventral midline of the developing Drosophila embryo, respectively. Here, evidence is presented that the sog primary enhancer also has both activities, with the late enhancer activity dependent on the early activity. Computational analyses showed that the sog primary enhancer contains five Dorsal (Dl)-, four Zelda (Zld)-, three Bicoid (Bcd)-, and no Single-minded (Sim)-binding sites. In contrast to many ventral midline enhancers, the primary enhancer can direct lacZ expression in the ventral midline as well as in the neurogenic ectoderm without a canonical Simbinding site. Intriguingly, the impaired transcriptional synergy between Dl and either Zld or Bcd led to aberrant and abolished lacZ expression in the neurogenic ectoderm and in the ventral midline, respectively. These findings suggest that the two enhancer activities of the sog primary enhancer are functionally consolidated and geographically inseparable.

• BMB Reports
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• 제48권10호
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• pp.589-594
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• 2015

The shadow enhancer of the short gastrulation (sog) gene directs its sequential expression in the neurogenic ectoderm and the ventral midline of the developing Drosophila embryo. Here, we characterize three unusual features of the shadow enhancer midline activity. First, the minimal regions for the two different enhancer activities exhibit high overlap within the shadow enhancer, meaning that one developmental enhancer possesses dual enhancer activities. Second, the midline enhancer activity relies on five Single-minded (Sim)-binding sites, two of which have not been found in any Sim target enhancers. Finally, two linked Dorsal (Dl)- and Zelda (Zld)-binding sites, critical for the neurogenic ectoderm enhancer activity, are also required for the midline enhancer activity. These results suggest that early activation by Dl and Zld may facilitate late activation via the noncanonical sites occupied by Sim. We discuss a model for Zld as a pioneer factor and speculate its role in midline enhancer activity.

• Journal of Acupuncture Research
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• 제19권4호
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• pp.140-151
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• 2002

This study was carried to identify whether acupuncture at several meridian points can affect the human anesthesia or not through the analysis of serum intermediates, such as, melatonin, serotonin, TSH, cortisol in human serum. We investigated the effect of accupuncture on the change of serum intermediates which have frequently been made a subject of discussion owing to its hyponic and sedative properties in practice. The result obtained were as follow ; 1. Serum melatonin levels was hardly affected at 40 minutes after general acupuncture on Hab-Gog, Nae-Gwan, and the other meridian points as compared with that($5.64{\pm}1.02pg/ml$) of resting control group. But on day 1 after acupuncture, these tend to increase significantly ($7.95{\pm}2.05pg/ml$ to $8.21{\pm}1.57pg/ml$, p <0.05). On the other hand, control group under sleep showed the serum level of $7.39{\pm}1.03pg/ml$. Additional acupuncture at In-Dang induced the increased melatonin level, especially at 40minutes after acupuncture(p <0.05). 2. Serotonin level in resting control serum recorded $51.14{\pm}8.17{\mu}g/ml$. Acupuncture at determined meridian point intend to increase average level of serotonin, but not significant. However observation on day 1 after acupuncture at Hab-Gwan meridian point and under sleeping showed the significantly increased serotonin level, $74.05{\pm}35.83{\mu}g/ml$, $70.44{\pm}13.08{\mu}g/ml$, respectively. 3. TSH level in intact human serum recorded $0.85{\pm}0.24{\mu}IU/ml$. and the serum exposed to acupuncture showed the irregular pattern of TSH level in a mean range of $0.76{\pm}0.44{\mu}IU/ml$ to $1.06{\pm}0.38{\mu}IU/ml$, regardless of meridian point or time after acupuncture. 4. The values of serum cortisol in control group were $11.76{\pm}2.06{\mu}g/dl$ in resting, $7.51{\pm}2.85{\mu}/dl$ under sleep, respectively. The level of serum cortisol in 40minutes after acupuncture was markedly (p <0.05) reduced to the serum levels of $5.65{\pm}2.58{\mu}g/dl$ (Hab-Gog, Nae-Gwan), $7.58{\pm}3.21{\mu}g/dl$ (Gyo-Gam, Sin-Mun besides Hab-Gog, Nae-Gwan), $6.43{\pm}3.54{\mu}g/dl$ (In-Dang besides to Hab-Gog, Nae-Gwan), respectively, as compared with control, intending to increase a little on day 1 after acupuncture. From the above results, the analysis of serum intermediates suggest that acupunctuation at meridian points applied to at this study act upon the phase of light anaesthesia or hypnosis, at the same time affect pituitary-adrenal axis rather than hypothalamus-pituitary axis in the secretary system of hormone and also don't affect dorsal-raphe nucleus according to the observation of change transition in serum intermediates, such as TSH, serotonin, and cortisol.

• The Korean Journal of Pain
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• 제19권1호
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• pp.33-44
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• 2006

Background: Peripheral nerve injury leads to neuropathic pain, including mechanical hyperalgesia (MH). Nerve discharges produced by an injury to the primary afferents cause the release of glutamate from both central and peripheral terminals. While the role of centrally released glutamate in MH has been well studied, relatively little is known about its peripheral role. This study was carried out to determine if the peripherally conducting nerve impulses and peripheral glutamate receptors contribute to the generation of neuropathic pain. Methods: Rats that had previously received a left L5 dorsal rhizotomy were subjected to a spinal nerve lesion (SNL) or brief electrical stimulation (ES, 4 Hz pulses for 5 min) of the left L5 spinal nerve. The paw withdrawal threshold (PWT) to von Frey filaments was measured. The effects of an intraplantar (i.pl.) injection of a glutamate receptor (GluR) antagonist or agonist on the changes in the SNL- or ES-produced PWT was investigated. Results: SNL produced MH, as evidenced by decrease in the PWT, which lasted for more than 42 days. ES also produced MH lasting for 7 days. MK-801 (NMDAR antagonist), DL-AP3 (group-I mGluR antagonist), and APDC (group-II mGluR agonist) delayed the onset of MH when an i.pl. injection was given before SNL. The same application blocked the onset of ES-induced MH. NBQX (AMPA receptor antagonist) had no effect on either the SNL- or ES-induced onset of MH. When drugs were given after SNL or ES, MK-801 reversed the MH, whereas NBQX, DL-AP3, and APDC had no effect. Conclusions: Peripherally conducting impulses play an important role in the generation of neuropathic pain, which is mediated by the peripheral glutamate receptors.

• The Korean Journal of Physiology and Pharmacology
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• 제13권3호
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• pp.209-214
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• 2009

The striatum receives glutamatergic afferents from the cortex and thalamus, and these synaptic transmissions are mediated by ${\alpha}$-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and N-methyl D-aspartate (NMDA) receptors. The purpose of this study was to characterize glutamate receptors by analyzing NMDA/AMPA ratio and rectification of AMPA and NMDA excitatory postsynaptic currents (EPSCs) using a whole-cell voltage-clamp method in the dorsal striatum. Receptor antagonists were used to isolate receptor or subunit specific EPSC, such as (DL)-2-amino-5-phosphonovaleric acid (APV), an NMDA receptor antagonist, ifenprodil, an NR2B antagonist, CNQX, an AMPA receptor antagonist and IEM-1460, a GluR2-lacking AMPA receptor blocker. AMPA and NMDA EPSCs were recorded at - 70 and + 40 mV, respectively. Rectification index was calculated by current ratio of EPSCs between + 50 and - 50 mV. NMDA/AMPA ratio was 0.20${\pm}$0.05, AMPA receptor ratio of GluR2-lacking/GluR2-containing subunit was 0.26${\pm}$0.05 and NMDA receptor ratio of NR2B/NR2A subunit was 0.32${\pm}$0.03. The rectification index (control 2.39${\pm}$0.27) was decreased in the presence of both APV and combination of APV and IEM-1460 (1.02${\pm}$0.11 and 0.93${\pm}$0.09, respectively). These results suggest that the major components of the striatal glutamate receptors are GluR2-containing AMPA receptors and NR2A-containing NMDA receptors. Our results may provide useful information for corticostriatal synaptic transmission and plasticity studies.

• The Korean Journal of Physiology and Pharmacology
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• 제12권6호
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• pp.293-297
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• 2008

The effect of forskolin on corticostriatal synaptic transmission was examined by recording excitatory postsynaptic currents (EPSCs) in rat brain slices using the whole-cell voltage-clamp technique. Forskolin produced a dose-dependent increase of corticostriatal EPSCs (1, 3, 10, and $30{\mu}M$) immediately after its treatment, and the increase at 10 and $30{\mu}M$ was maintained even after its washout. When the brain slices were pre-treated with (DL)-2-amino-phosphonovaleric acid (AP-V, $100{\mu}M$), an NMDA receptor antagonist, the acute effect of forskolin ($10{\mu}M$) was blocked. However, after washout of forskolin, an increase of corticostriatal EPSCs was still observed even in the presence of AP-V. When KT 5720 ($5{\mu}M$), a protein kinase A (PKA) inhibitor, was applied through the patch pipette, forskolin ($10{\mu}M$) increased corticostriatal EPSCs, but this increase was not maintained. When forskolin was applied together with AP-V and KT 5720, both the increase and maintenance of the corticostriatal EPSCs were blocked. These results suggest that forskolin activates both NMDA receptors and PKA, however, in a different manner.

• International Journal of Oral Biology
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• 제41권3호
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• pp.141-147
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• 2016

Reactive oxygen species (ROS) and nitrogen species (RNS) are both important signaling molecules involved in pain transmission in the dorsal horn of the spinal cord. Xanthine oxidase (XO) is a well-known enzyme for the generation of superoxide anions ($O_2^{\bullet-}$), while S-nitroso-N-acetyl-DL-penicillamine (SNAP) is a representative nitric oxide (NO) donor. In this study, we used patch clamp recording in spinal slices of rats to investigate the effects of $O_2^{\bullet-}$ and NO on the excitability of substantia gelatinosa (SG) neurons. We also used confocal scanning laser microscopy to measure XO- and SNAP-induced ROS and RNS production in live slices. We observed that the ROS level increased during the perfusion of xanthine and xanthine oxidase (X/XO) compound and SNAP after the loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), which is an indicator of intracellular ROS and RNS. Application of ROS donors such as X/XO, ${\beta}-nicotinamide$ adenine dinucleotide phosphate (NADPH), and 3-morpholinosydnomimine (SIN-1) induced a membrane depolarization and inward currents. SNAP, an RNS donor, also induced membrane depolarization and inward currents. X/XO-induced inward currents were significantly decreased by pretreatment with phenyl N-tert-butylnitrone (PBN; nonspecific ROS and RNS scavenger) and manganese(III) tetrakis(4-benzoic acid) porphyrin (MnTBAP; superoxide dismutase mimetics). Nitro-L-arginine methyl ester (NAME; NO scavenger) also slightly decreased X/XO-induced inward currents, suggesting that X/XO-induced responses can be involved in the generation of peroxynitrite ($ONOO^-$). Our data suggest that elevated ROS, especially $O_2^{\bullet-}$, NO and $ONOO^-$, in the spinal cord can increase the excitability of the SG neurons related to pain transmission.