• Title/Summary/Keyword: Cx32

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Molecular Cloning and Nucleotide Sequence of Connexin 35 cDNA in the Ovary from the Sweetfish, Plecoglossus altivelis (은어, Plecoglossus altivelis 난소에서 발현하는 Connexin 35 cDNA의 해석)

  • Choi, Cheol-Young;Chang, Young-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.565-571
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    • 2000
  • Mixed primers based on the high sequence homology of selected regions of known connexins (Cxs) was used for PCR reaction. A full-length connexin cDNA of sweetfish (Plecoglossus altivelis) was cloned by rapid amplification of cDNA 5'and (5'RACE) and 3'RACE method. When compared to other known Cx sequences, homology of sweetfish Cx cDNA to Atlantic croaker, Mycropogonias undulatus Cx32.7, bovine, Bos taurus Cx44 and Atlantic croaker Cx32.2 were $63.8{\%},\;61.6{\%}\;and\;56.7{\%}$, respectively. This cDNA encoded 308 amino acids (35,028 dalton) and named as sweetfish Cx35. Hydropathicity analysis of predicted amino acid sequences indicated that sweetfish Cx35 have four major hydrophobic regions and four major hydrophilic regions, suggesting its topology is similar to that of known Cxs. The presence of a tfical Cx consensus sequences were identified in each of the extracellular loops (first loop and second loop).

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Changes in Expression of Connexin Isoforms in the Caudal Epididymis of Adult Sprague-Dawley Rats exposed to Estradiol Benzoate or Flutamide at the Neonatal Age

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.20 no.3
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    • pp.237-245
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    • 2016
  • Direct communication between neighboring cells via gap junction in tissue is important for maintenance and regulation of its physiological functions. Each epididymal region has different composition of cell types. It is well recognized that the epididymis is a steroid hormone-responsive tissue. The present study was designed to determine the effect of estradiol benzoate (EB) or flutamide exposured at the early postnatal age on the expression of connexin (Cx) isoforms in the caudal epididymis. The EB or flutamide was subcutaneously administrated to male Spragure Dawley rat at 7 days of age, and expressional changes of Cx isoforms in the adult corpus epididymis were determined by quantitative real-time PCR. The treatment of low-dose EB resulted in decreases of Cx30.3, Cx31.1, Cx37, and Cx45 expression but caused an increase of Cx32 expression. Exposure to high-dose EB led into expressional increases of Cx31, Cx31.1, Cx32, Cx40, and Cx43, even though a decrease of Cx37 expression was found with a high-dose EB treatment. A low-dose flutamide induced increases of Cx31, Cx31.1, Cx32, and Cx43 expression but a decrease of Cx37 expression. Expression of most Cx genes were significantly increased by a high-dose flutamide, while no expressional change of Cx26 and Cx40 was detected by a high-dose flutamide. These results indicate that expression of Cx isoforms in the caudal epididymis is altered by exposure to steroidal compounds at the prepubertal age. It is suggested that a contact with environmental exogenous materials during the early postnatal period would lead to alteration of epididymal functions at the adult.

Aberrant Expression of Cx Isoforms in the Adult Caput Epididymis exposed to Estradiol Benzoate or Flutamide at the Weaning

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.21 no.4
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    • pp.379-389
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    • 2017
  • Connexin (Cx) involves in the regulation of various physiological functions of tissue by forming a channel, a gap junction which allows direct cell-cell communication, between adjacent cells. The effect of a single subcutaneous treatment of estradiol benzoate (EB) or flutamide (Flu) at the weaning age on the expression of Cx isoforms in the adult caput epididymis was evaluated in this research. Using quantitative real-time PCR analysis, a low-dose of EB [$0.015{\mu}g/kg$ body weight (BW)] caused significant decreases of Cx30.3, Cx32, Cx40, Cx43, and Cx45 mRNA levels and no change of Cx26, Cx31, Cx31.1, Cx37 transcript levels. The treatment of a high-dose EB ($1.5{\mu}g/kg\;BW$) resulted in reduced expression of Cx30.3, Cx31, Cx43, and Cx45 but increased expression of Cx37 and Cx40. Expression of all Cx isoforms examined, except Cx31, was significantly increased by the treatment of a low-dose Flu ($500{\mu}g/kg\;BW$). However, the treatment of a high-dose Flu (5 mg/kg BW) led significant expressional suppression of Cx30.3, Cx31, Cx31.1, Cx32, Cx40, Cx43, and Cx45 but an increase of Cx37 transcript level. With the comparison of previous findings, the expression of Cx isoforms in the adult epididymis after the exposure to EB or Flu is likely differentially regulated in regional-specific and/or exposed postnatal age-specific manner.

Single Channel Analysis of Xenopus Connexin 38 Hemichannel (제노푸스 Cx38 세포막채널의 단일채널분석)

  • Cheon, Mi-Saek;Oh, Seung-Hoon
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1517-1522
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    • 2007
  • Gap junction channels formed by two adjacent cells allow the passage of small molecules up to ${\sim}\;1\;kDa$ between them. Hemichannel (connexon or half of gap junction) also behaves as a membrane channel like sodium or potassium channels in a single cell membrane. Among 26 types of connexin (Cx), $Cx32^*43E1$ (a chimera in which the first extracellular loop of Cx32 has been replaced with that of Cx43), Cx38, Cx46, and Cx50 form functional hemichannels as well as gap junction channels. Although it is known that Xenopus oocytes express endogenous connexin 38 (Cx38), its biophysical characteristics at single channel level are poorly understood. In this study, we performed single channel recordings from single Xenopus oocytes to acquire the biophysical properties of Cx38 including voltage-dependent gating and permeation (conductance and selectivity). The voltage-dependent fast and slow gatings of Cx38 hemichannel are distinct. Fast gating events occur at positive potentials and their open probabilities are low. In contrast, slow gatings dominate at negative potentials with high open probabilites. Based on hi-ionic experiments, Cx38 hemichannel is anion-selective. It will be interesting to test whether charged amino acid residues in the amino terminus of Cx38 are responsible for voltage gatings and permeation.

Modification of Gene Expression of Connexins in the Rat Corpus Epididymis by Estradiol Benzoate or Flutamide Exposure at the Early Neonatal Age

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.19 no.2
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    • pp.69-77
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    • 2015
  • Cell-cell direct communication through channel-forming molecules, connexin (Cx), is essential for a tissue to exchange signaling molecules between neighboring cells and establish unique functional characteristics during postnatal development. The corpus epididymis is a well-known androgen-responsive tissue and involves in proper sperm maturation. In the present research, it was attempted to determine if expression of Cx isoforms in the corpus epididymis in the adult is modulated by exposure to estrogenic or anti-androgenic compound during the early postnatal period. The neonatal male rats at 7 days of age were subcutaneously injected by estradiol benzoate (EB) at low-dose ($0.015{\mu}g/kg$ body weight) or high-dose ($1.5{\mu}g/kg$ body weight) or flutamide (Flu) at low-dose ($500{\mu}g/kg$ body weight) or high-dose (50 mg/kg body weight). The corpus epididymis collected at 4 months of age was subjected to evaluate expressional changes of Cx isoforms by quantitative real-time PCR. Treatment of low-dose EB resulted in increases of Cx32, Cx37, and Cx45 transcript levels, while exposure to high-dose EB decreased expression of Cx26, Cx30.3, Cx31, Cx31.1, Cx32, Cx40, Cx43, and Cx45. Treatments of Flu caused significant decreases of expression of all examined Cx isoforms, except Cx37 and Cx43 shown no expressional change with high-dose Flu treatment. These findings imply that expression of most Cx isoforms present in the corpus epididymis would be transcriptionally regulated by actions of androgen and/or estrogen during postnatal period.

Examination of the Presence and Differential Expression of Connexin Isoforms in the Urinary Bladder of the Male Rat During Postnatal Period

  • Lee, Ki-Ho
    • Journal of Animal Science and Technology
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    • v.54 no.6
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    • pp.419-425
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    • 2012
  • The coordinated action of the detrusor muscle cells in the urinary bladder is governed by cell-cell communication through gap junction, consisted of connexin (Cx) molecules. Even though a number of researches have been mostly focused on expressional changes of a few Cx isoforms in clinically dysfunctional condition of the bladder, less attention has been paid for investigation of Cx isoforms present in the bladder. Using real-time PCR analysis, the present study examined Cx isoforms expressing in the male rat bladder during postnatal period. Also, expressional patterns of Cx isoforms were evaluated in the bladder at different postnatal ages. Of a total of 13 Cx isoforms tested in the present study, we were able to detect mRNAs of 6 Cx isoforms in the rat urinary bladder, including Cxs 31, 31.1, 32, 37, 40, and 45. The transcript levels of Cxs 31, 31.1, 37, 40, and 45 were gradually increased from 1 week of age until 25 days of age, followed by transient decreases at 45 days of age. However, abundance of Cx32 transcript was drastically increased at 15 days of age, followed by a sharp drop at 45 days of age. These results indicate that differential expression of Cx isoforms in the bladder during postnatal development would be necessary for maintaining proper function of the bladder. A question remains to be answered if significant decreases of transcript levels of some Cx isoforms at the elderly are associated with age-dependent dysfunction of the bladder.

Connexin32 inhibits gastric carcinogenesis through cell cycle arrest and altered expression of p21Cip1 and p27Kip1

  • Jee, Hyang;Lee, Su-Hyung;Park, Jun-Won;Lee, Bo-Ram;Nam, Ki-Taek;Kim, Dae-Yong
    • BMB Reports
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    • v.46 no.1
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    • pp.25-30
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    • 2013
  • Gap junctions and their structural proteins, connexins (Cxs), have been implicated in carcinogenesis. To explore the involvement of Cx32 in gastric carcinogenesis, immunochemical analysis of Cx32 and proliferation marker Ki67 using tissue-microarrayed human gastric cancer and normal tissues was performed. In addition, after Cx32 overexpression in the human gastric cancer cell line AGS, cell proliferation, cell cycle analyses, and $p21^{Cip1}$ and $p27^{Kip1}$ expression levels were examined by bromodeoxyuridine assay, flow cytometry, real-time RT-PCR, and western blotting. Immunohistochemical study noted a strong inverse correlation between Cx32 and Ki67 expression pattern as well as their location. In vitro, overexpression of Cx32 in AGS cells inhibited cell proliferation significantly. $G^1$ arrest, up-regulation of cell cycle-regulatory proteins $p21^{Cip1}$ and $p27^{Kip1}$ was also found at both mRNA and protein levels. Taken together, Cx32 plays some roles in gastric cancer development by inhibiting gastric cancer cell proliferation through cell cycle arrest and cell cycle regulatory proteins.

Expressional Modulation of Connexin Isoforms in the Initial Segment of Male Rat treated with Estradiol Benzoate or Flutamide

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.18 no.4
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    • pp.293-300
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    • 2014
  • Direct cell-cell communication through connexin (Cx) complexes is a way to achieve functional accordance of cells within a tissue or an organ. The initial segment (IS), a part of the epididymis, plays important roles in sperm maturation. Steroid hormones influence on expression of a number of genes in the IS of adult animals. However, developmental effect of sex hormones on the gene expression in the IS has not been examined. In this study, estradiol benzoate (EB, an estrogen agonist) or flutamide (Flu, an androgen antagonist) was exogenously administrated at 1 week of postnatal age, and expressional changes of Cx genes in the IS were determined at 4 months of age by a quantitative real-time PCR analysis. Treatment of EB at $0.015{\mu}g/kg$ body weight (BW) increased expression of Cx30.3, 31.1, and 43 genes. However, treatment of 1.5 mg EB/kg BW resulted in expressional decreases of Cx31, 32, and 45 genes and caused increases of Cx30.3 and 43 gene expression. Significant decreases of Cx31, 31.1, 32, 37, and 45 gene expression were detected with a treatment of $500{\mu}g\;Flu/kg$ BW, while expression of Cx43 gene was significantly increased with a treatment of $500{\mu}g\;Flu/kg$ BW. A treatment of $50{\mu}g\;Flu/kg$ BW led to significant increases of Cx30.3, 32, 37, 40, and 43 gene expression. These findings imply that exogenous exposure of steroidal hormones during the early developmental period would result in aberrant expression of Cx genes in the adult IS.

Expressional Patterns of Connexin Isoforms in the Rat Epididymal Fat during Postnatal Development

  • Lee, Ki-Ho;Kim, Nan Hee
    • Development and Reproduction
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    • v.22 no.1
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    • pp.29-38
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    • 2018
  • In the multicellular tissue, cell-cell interaction is important for a precise control of its function. The exchange of signaling molecules between adjacent cells via connexon allows the functional harmony of cells in the tissue. The present research was to determine the presence and expressional patterns of connexin (Cx) isoforms in the rat epididymal fat during postnatal development using quantitative real-time polymerase chain reaction (PCR) analysis. Of 13 Cx isoforms examined, expression of 11 Cx isoforms in the epididymal fat during postnatal development was detected. These Cx isoforms include Cx26, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx43, Cx45, and Cx50. Expressional levels of all Cx isoforms at 1 and 2 years of age were significantly higher than those at the early postnatal ages, such as 7 days, 14 days, and 24 days of ages. Except Cx33 and Cx43, the transcript levels of rest Cx isoforms at 1 year of age were significantly lower than that at 2 years of age. In addition, expressional patterns of Cx isoforms between 7 days and 5 months of ages generally varied according to the isoform. The existence of various Cx isoforms in the rat epididymal fat has been identified and expression of each Cx isoform in the epididymal fat during postnatal development has shown a particular pattern, distinguishable from the others. To our knowledges, this is the first report showing expressional patterns of Cx isoforms at transcript level in the epididymal fat at various postnatal ages.

Development of the Q-methodology Integrated Customer Experience Management Process Based on Customer Journey Map for Improving Customer Experience: CX-Q (고객경험 개선을 위한 고객여정지도 기반 Q-방법론 통합 고객경험관리 프로세스 제안: CX-Q)

  • Yu, Seong Hun;Park, Do Hyung
    • The Journal of Information Systems
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    • v.32 no.1
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    • pp.201-221
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    • 2023
  • Purpose Customers consider the overall experience with the company as important as the quality of the product, and companies are also paying attention to creating long-term relationships with customers through optimal customer experiences. In this study, we propose a customer experience management process called 'CX-Q', which combines customer journey map and Q-methodology to understand the importance of customer experience based on the overall customer experience. Design/methodology/approach. CX-Q is a process that combines Q-methodology and customer journey maps, allowing stakeholders to explore and improve customer experiences at each contact point while engaging with brands, products, and services. It also enables them to derive customer experience insights and important management points for each segment. To demonstrate the usefulness of the proposed CX-Q, this study analyzed the experience of customers who used the Airbnb travel platform service as an example, applying the CX-Q process. Findings A total of four customer segments were derived, and it was found that each segment valued different attributes during the customer journey stage. The customer experience analysis using the CX-Q process proposed in this study is expected to help understand customers in more detail and assist in managing and improving customer experience.