• Title/Summary/Keyword: Light-dependent Expression

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A Novel Phototransduction Pathway in the Pineal Gland and Retina

  • Okano, Toshiyuki;Kasahara, Takaoki;Fukada, Yoshitaka
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.246-248
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    • 2002
  • Light is a major environmental signal for entrainment of the circadian clock, but little is known about the phototransduction pathway triggered by light-activation of photoreceptive molecule(s) responsible for the phase shift of the clock in vertebrates. The chicken pineal gland and retina contain the autonomous circadian oscillators together with the photic entrainment pathway, and hence they provide useful experimental model for the clock system. We previously demonstrated the expression and light-dependent activation of rod-type transducin $\alpha$-subunit (Gtl$\alpha$) in the chicken pineal gland. It is unlikely, however, that the pineal Gt$_1$$\alpha$ plays a major role in the photic entrainment, because the light-induced phase shift is unaffected by bloking the signaling function of Gt$_1$$\alpha$. Here, we show the expression of G 11 $\alpha$, an $\alpha$-subunit of another heterotrimeric G-protein, in the chicken pineal gland and retina by cDNA cloning, Northern blot and Western blot analyses. GIl$\alpha$-immunoreactivity was colocalized with pinopsin in the chicken pineal cells and it was found predominantly at the outer segments of photoreceptor cells in the retinal sections, suggesting functional coupling of G11 $\alpha$ with opsins in the both the tissues. By coimmunoprecipitation experiments using the retina, we showed the light- and GTP-dependent interaction between rhodopsin and G11 $\alpha$. Upon ectopic expression of a Gq/ 11-coupled receptor in cultured pineal cells, pharmacological (non-photic) activation of endogenous G11 induced phase-dependent phase shifts of the melatonin rhythm in a manner very similar to the effect of light. These results suggested opsin-G11 pathway contributing to the photic entrainment of the circadian clock.

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Molecular Mechanism of Photic-Entrainment of Chicken Pineal Circadian Clock

  • Okano, Toshiyuki;Fukada, Yoshitaka
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.25-28
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    • 2002
  • The chicken pineal gland has been used for studies on the circadian clock, because it retains an intracellular phototransduction pathway regulating the phase of the intrinsic clock oscillator. Previously, we identified chicken clock genes expressed in the gland (cPer2, cPer3, cBmal1, cBmal2, cCry1, cCry2, and cClock), and showed that a cBMALl/2-cCLOCK heteromer acts as a regulator transactivating cPer2 gene through the CACGTG E-box element found in its promoter. Notably, mRNA expression of cPer2 gene is up-regulated by light as well as is driven by the circadian clock, implying that light-dependent clock resetting may involve the up-regulation of cPer2 gene. To explore the mechanism of light-dependent gene expression unidentified in animals, we first focused on pinopsin gene whose mRNA level is also up-regulated by light. A pinopsin promoter was isolated and analyzed by transcriptional assays using cultured chicken pineal cells, resulting in identification of an 18-bp light-responsive element that includes a CACGTG E-box sequence. We also investigated a role of mitogen-activated protein kinase (MAPK) in the clock resetting, especially in the E-box-dependent transcriptional regulation, because MAPK is phospholylated (activated) in a circadian manner and is rapidly dephosphorylated by light in the gland. Both pulldown analysis and kinase assay revealed that MAPK directly associates with BMAL1 to phosphorylate it at several Ser/Thr residues. Transcriptional analyses implied that the MAPK-mediated phosphorylation may negatively regulate the BMAL-CLOCK-dependent transactivation through the E-box. These results suggest that the CACGTG E-box serves not only as a clock-controlled element but also as a light-responsive element.

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MET1-Dependent DNA Methylation Represses Light Signaling and Influences Plant Regeneration in Arabidopsis

  • Shim, Sangrea;Lee, Hong Gil;Seo, Pil Joon
    • Molecules and Cells
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    • v.44 no.10
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    • pp.746-757
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    • 2021
  • Plant somatic cells can be reprogrammed into a pluripotent cell mass, called callus, which can be subsequently used for de novo shoot regeneration through a two-step in vitro tissue culture method. MET1-dependent CG methylation has been implicated in plant regeneration in Arabidopsis, because the met1-3 mutant exhibits increased shoot regeneration compared with the wild-type. To understand the role of MET1 in de novo shoot regeneration, we compared the genome-wide DNA methylomes and transcriptomes of wildtype and met1-3 callus and leaf. The CG methylation patterns were largely unchanged during leaf-to-callus transition, suggesting that the altered regeneration phenotype of met1-3 was caused by the constitutively hypomethylated genes, independent of the tissue type. In particular, MET1-dependent CG methylation was observed at the blue light receptor genes, CRYPTOCHROME 1 (CRY1) and CRY2, which reduced their expression. Coexpression network analysis revealed that the CRY1 gene was closely linked to cytokinin signaling genes. Consistently, functional enrichment analysis of differentially expressed genes in met1-3 showed that gene ontology terms related to light and hormone signaling were overrepresented. Overall, our findings indicate that MET1-dependent repression of light and cytokinin signaling influences plant regeneration capacity and shoot identity establishment.

CLONING AND LIGHT-DEPENDENT EXPRESSION OF A cDNA FOR PEA CYTOSOLIC FRUCTOSE-1,6-BISPHOSPHATASE

  • Son, Tae-Jong;Hahn, Tae-Ryong
    • Journal of Photoscience
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    • v.4 no.3
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    • pp.141-145
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    • 1997
  • Polymerase chain reaction(PCR) was conducted with a pea cDNA library using two primers synthesized from homology analysis of amino acid sequences for animal and plant cytosolic FBPases. A PCR product with 650 bp long was cloned into pGEM-T vector and sequenced. The deduced amino acid sequence of the cDNA fragment was 98, 91, and 85% homologous with those of cytosolic FBPases from spinach, sugarbeet, and sugarcane, respectively. It was 51% homologous with amino acid sequence of FBPase from pea chloroplasts. Northern blot analysis was proceeded with the cDNA clone resulting that 1.2 kb transcript was highly expressed in light-grown pea leaves but almost not expressed in dark-grown etiolated pea seedlings. When peas grown in the light for 10 days were transferred to darkness, the transcript was gradually decreased with dark treatment, indicating that the expression of the enzyme was induced by continuous white light but suppressed by dark treatment. Pea cytosolic FBPase was highly expressed in leaves with trace amounts in stems. but almost not expressed in roots.

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Altered Gene Expression in Cerulein-Stimulated Pancreatic Acinar Cells: Pathologic Mechanism of Acute Pancreatitis

  • Yu, Ji-Hoon;Lim, Joo-Weon;Kim, Hye-Young
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.6
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    • pp.409-416
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    • 2009
  • Acute pancreatitis is a multifactorial disease associated with the premature activation of digestive enzymes. The genes expressed in pancreatic acinar cells determine the severity of the disease. The present study determined the differentially expressed genes in pancreatic acinar cells treated with cerulein as an in vitro model of acute pancreatitis. Pancreatic acinar AR42J cells were stimulated with $10^{-8}$ M cerulein for 4 h, and genes with altered expression were identified using a cDNA microarray for 4,000 rat genes and validated by real-time PCR. These genes showed a 2.5-fold or higher increase with cerulein: lithostatin, guanylate cyclase, myosin light chain kinase 2, cathepsin C, progestin-induced protein, and pancreatic trypsin 2. Stathin 1 and ribosomal protein S13 showed a 2.5-fold or higher decreases in expression. Real-time PCR analysis showed time-dependent alterations of these genes. Using commercially available antibodies specific for guanylate cyclase, myosin light chain kinase 2, and cathepsin C, a time-dependent increase in these proteins were observed by Western blotting. Thus, disturbances in proliferation, differentiation, cytoskeleton arrangement, enzyme activity, and secretion may be underlying mechanisms of acute pancreatitis.

Optical Modeling for Polarization-dependent Optical Power Dissipation of Thin-film Organic Solar Cells at Oblique Incidence

  • Kim, Jungho;Jung, Sungyeop;Jeong, Inkyung
    • Journal of the Optical Society of Korea
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    • v.16 no.1
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    • pp.6-12
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    • 2012
  • We present the optical models and calculation results of thin-film organic solar cells (OSCs) at oblique incidence of light, using the transfer matrix method. The simple expression for the optical power dissipation is derived at oblique incidence for s- and p-polarized light. The spatial distribution of the electric field intensity, the optical power density, and the optical power dissipation are calculated in both s- and p-polarized light with respect to the incidence angle. We identify how the light absorption efficiency for p-polarized light becomes relatively larger than that for s-polarized light as the incidence angle increases.

The Phenotype of the Soybean Disease-Lesion Mimic (dlm) Mutant is Light-Dependent and Associated with Chloroplast Function

  • Kim, Byo-Kyong;Kim, Young-Jin;Paek, Kyoung-Bee;Chung, Jong-Il;Kim, Jeong-Kook
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.395-401
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    • 2005
  • The dlm (disease lesion mimic) mutant of soybean (Glycine max L. Merr) shows the similar lesion of a soybean disease caused by a fungus, Corynespora cassilcola. The lesion was examined at cellular and molecular level. Trypan blue staining result indicated that cell death was detectable in the entire region of leaves excluding veins when the lesions had already been developed. We found that the mesophyll cells of palisade layer in the dim mutant appeared to be wider apart from each other. The chloroplasts of the dim mutant cells contained bigger starch granules than those in normal plants. We also found that the lesion development of dlm plant was light-dependent and the starch degradation during the dark period of diurnal cycle was impaired in the mutant. Three soybean pathogenesis-related genes, PR-1a, PR-4, and PR-10, were examined for their expression patterns during the development of disease lesion mimic. The expression of all three genes was up-regulated to some extent upon the appearance of the disease lesion mimic. Although the exact function of DLM protein remains elusive, our data would provide some insight into mechanism underling the cell death associated with the dim mutation.

Ameliorative effects of atractylodin on intestinal inflammation and co-occurring dysmotility in both constipation and diarrhea prominent rats

  • Yu, Changchun;Xiong, Yongjian;Chen, Dapeng;Li, Yanli;Xu, Bin;Lin, Yuan;Tang, Zeyao;Jiang, Chunling;Wang, Li
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.1
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    • pp.1-9
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    • 2017
  • Intestinal disorders often co-occur with inflammation and dysmotility. However, drugs which simultaneously improve intestinal inflammation and co-occurring dysmotility are rarely reported. Atractylodin, a widely used herbal medicine, is used to treat digestive disorders. The present study was designed to characterize the effects of atractylodin on amelioration of both jejunal inflammation and the co-occurring dysmotility in both constipation-prominent (CP) and diarrhea-prominent (DP) rats. The results indicated that atractylodin reduced proinflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 in the plasma and inhibited the expression of inflammatory mediators iNOS and NF-kappa B in jejunal segments in both CP and DP rats. The results indicated that atractylodin exerted stimulatory effects and inhibitory effects on the contractility of jejunal segments isolated from CP and DP rats respectively, showing a contractile-state-dependent regulation. Atractylodin-induced contractile-state-dependent regulation was also observed by using rat jejunal segments in low and high contractile states respectively (5 pairs of low/high contractile states). Atractylodin up-regulated the decreased phosphorylation of 20 kDa myosin light chain, protein contents of myosin light chain kinase (MLCK), and MLCK mRNA expression in jejunal segments of CP rats and down-regulated those increased parameters in DP rats. Taken together, atractylodin alleviated rat jejunal inflammation and exerted contractile-state-dependent regulation on the contractility of jejunal segments isolated from CP and DP rats respectively, suggesting the potential clinical implication for ameliorating intestinal inflammation and co-occurring dysmotility.

Transcriptome Analysis of Long-Term Exposure to Blue Light in Retinal Pigment Epithelial Cells

  • Jin, Hong Lan;Jeong, Kwang Won
    • Biomolecules & Therapeutics
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    • v.30 no.3
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    • pp.291-297
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    • 2022
  • Dry age-related macular degeneration (AMD) is a type of progressive blindness that is primarily due to dysfunction and the loss of retinal pigment epithelium (RPE). The accumulation of N-retinylidene-N-retinylethanolamine (A2E), a by-product of the visual cycle, causes RPE and photoreceptor degeneration that impairs vision. Genes associated with dry AMD have been identified using a blue light model of A2E accumulation in the retinal pigment epithelium and transcriptomic studies of retinal tissue from patients with AMD. However, dry macular degeneration progresses slowly, and current approaches cannot reveal changes in gene transcription according to stages of AMD progression. Thus, they are limited in terms of identifying genes responsible for pathogenesis. Here, we created a model of long-term exposure to identify temporally-dependent changes in gene expression induced in human retinal pigment epithelial cells (ARPE-19) exposed to blue light and a non-cytotoxic dose of A2E for 120 days. We identified stage-specific genes at 40, 100, and 120 days, respectively. The expression of genes corresponding to epithelial-mesenchymal transition (EMT) during the early stage, glycolysis and angiogenesis during the middle stage, and apoptosis and inflammation pathways during the late stage was significantly altered by A2E and blue light. Changes in the expression of genes at the late stages of the EMT were similar to those found in human eyes with late-stage AMD. Our results provide further insight into the pathogenesis of dry AMD induced by blue light and a novel model in vitro with which relevant genes can be identified in the future.

Comparative Proteomic Analysis of Blue Light Signaling Components in the Arabidopsis Cryptochrome 1 Mutant

  • Phee, Bong-Kwan;Park, Sebyul;Cho, Jin-Hwan;Jeon, Jong-Seong;Bhoo, Seong Hee;Hahn, Tae-Ryong
    • Molecules and Cells
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    • v.23 no.2
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    • pp.154-160
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    • 2007
  • An Arabidopsis hy4 mutant that is specifically impaired in its ability to undergo blue light dependent photomorphogenesis was used to identify cryptochrome 1 signaling-related components. Proteomic analysis revealed about 205 differentially expressed protein spots in the blue light-irradiated hy4 mutant compared to the wild-type. The proteins corresponding to 28 up-regulated and 33 down-regulated spots were identified. Obvious morphological changes in the hy4 mutant were closely related to the expression of various transcription factors. Our findings suggest that blue light signals may be involved in many cellular processes including disease resistance and stress responses.