• 제목/요약/키워드: Aquaporin 1 (AQP1)

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Aquaporin 4 water channel 인산화에 의한 수분 투과도의 조절 (Phosphorylation of AQP4 Water Channel Regulates Water Permeability)

  • 박권희;정동근;정진섭;이재숙;예운해;서덕준;배혜란
    • 생명과학회지
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    • 제10권5호
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    • pp.456-466
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    • 2000
  • Aquaperin 4 (AQP4) is the mercurial water channel expressed abundantly in brain, especially the region related with cerebrospinal fluid reabsorption and osmoregulation. The primary structure of AQP4 water channel was elucidated but the molecular mechanism of AQP4 channel regulation is still unknown. To investigate the possible regulation of AQP4 water channel by phosphorylation via various protein kinases, osmotic water permeability of AQP4 expressed in Xenopus oocytes was measured by videomicroscopy technique. Forskolin (10 $\mu$M) did not affect osmotic water permeability of oocytes injected with AQP4 cRNA, excluding the regulation of AQP4 water cnannel by protein kinase A. Osmotic water permeability (P아래첨자) of AQP4-expressed oocytes was ingibited by the pretreatmeat of BAPTA/AM (up to 500$\mu$M), an intracellular Ca윗첨자 chelator, and calmidazolium (100$\mu$M), a specific Ca윗첨자/calmodulin antagonist, in a dose-dependent manner. The inhibition of osmotic water permeability (P아래첨자) by the calmidazolium treatment was completely reversed by the addition of calyculin A (0.1$\mu$M), a nonspecific phosphatase inhibitor. Phorbol 12-myristate 13-acetate (PMA), a protein kinase C activator, had biphasic effects on osmotic water permeability in AQP4 cRNA injected oocytes depending on its concentration; 21% increase by 100 nM PMA, 35% decrease by 1$\mu$M PMA. These effects were reversed with 2$\mu$M staurosporine, a nonspecific PKC inhibitor. These results suggest that phosphorylation of AQP4 water channel by Ca윗첨자/calmodulin kinase and protein kinase C might regulate the osmotic water permeability.

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Blunted Indomethacin-Induced Downregulation of Aquaporins by Nitric Oxide Synthesis Inhibition in Rats

  • You, Ju-Hee;Lee, Sung-Su;Bae, Eun-Hui;Ma, Seong-Kwon;Kim, Soo-Wan;Lee, Jong-Un
    • The Korean Journal of Physiology and Pharmacology
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    • 제10권4호
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    • pp.213-216
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    • 2006
  • The present study was aimed to determine whether nitric oxide (NO) plays a role in the regulation of aquaporin (AQP) channels in the kidney. Male Brattleboro rats ($250{\sim}300\;g$ body weight) were used. The experimental group was treated with $N^G$-nitro-L-arginine methyl ester (L-NAME, 100 mg/L drinking water) for 1 week, and cotreated with indomethacin (5 mg/kg, twice a day, i.p.) for the last two days. Control groups were treated with either L-NAME for 1 week, indomethacin for 2 days, or without any drug treatment. The abundance of AQP1, AQP2 and AQP3 proteins in the kidney was determined by Western blot analysis. Indomethacin downregulated AQP channels, whereas L-NAME by itself showed no significant effects on them. The indomethacin-induced downregulation of AQP2 and AQP3 was significantly blunted in L-NAME-treated rats, while that of AQP1 was not affected. These results suggest that endogenous NO, when stimulated, may downregulate AQP channels that are specifically regulated by AVP/cAMP pathway in the kidney.

왕불유행, 목통, 통초가 산후 생쥐의 유즙분비량과 유즙분비 관련 인자에 미치는 영향 (Effect of Melandrii Herba, Akebia Quinata Decaisne, and Tetrapanax Papyriferus on Milk Secretion and Lactation Related Factors in Postpartum Mice)

  • 이가위;이은희;이창현;김홍준
    • 대한한방부인과학회지
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    • 제31권2호
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    • pp.1-17
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    • 2018
  • Objectives: The purpose of this study is to investigate the effect of Melandrii Herba (MH), Akebia Quinata Decaisne (AQ), and Tetrapanax Papyriferus (TP) on milk secretion and aquaporin (AQP) expression in lactating mice. Methods: For the experiment, the mice were divided into three groups, which were orally administered MH (2,720 mg/kg), TP (400 mg/kg) and AQ (2,800 mg/kg) extracts respectively for 3 weeks from Day 1 after the birth, compared with the control group (C group), which was administered distilled water. A group consisted of six infantile mice per postpartum mouse. For comparison with the C group, non-pregnant SKH-1 mice were used as the virgin group. Results: 1. When it comes to the immunohistochemical staining for prolactin receptors in the mammary glands, the AQ and MH groups showed a strong immune response to the secretory epithelial cells constituting the mammary alveoli, while the TP group represented a weaker immune response. 2. In the immunohistochemical staining for AQP in the mammary glands, AQP1 showed a strong immune response in the walls of capillaries and venules around the mammary alveoli, and AQP3 in the epithelial cells constituting the mammary alveoli, and AQP5 in some tissues between the mammary alveoli. AQP1 was expressed in the order of TP group>AQ group=C group>MH group, and AQP3 was MH group and AQ group>TP group=C group, and AQP5 was MH group>C group>AQ group and TP group. 3. In the Western blot, AQP1 was expressed in the order of TP group>AQ group>C group>MH group, and AQP3 was MH group>AQ group>C group>TP roup, and AQP5 was MH group>TP Group>C group>AQ group. All of AQP1, 3, 5 expression were significantly higher in the C group than in the Virgin group. Conclusions: The administration of Akebia Quinata Decaisne, Tetrapanax Papyriferus and Melandrii Herba have the effect of improving prolactin levels in postpartum mice and increasing the expression of prolactin receptor and AQPs in the mammary glands, suggesting that lactation might be enhanced by the development of the mammary glands.

Temporal Aquaporin 11 Expression and Localization during Preimplantation Embryo Development

  • Park, Jae-Won;Cheon, Yong-Pil
    • 한국발생생물학회지:발생과생식
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    • 제19권1호
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    • pp.53-60
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    • 2015
  • Environmental conditions during early mammalian embryo development are critical and some adaptational phenomena are observed. However, the mechanisms underlying them remain largely masked. Previously, we reported that AQP5 expression is modified by the environmental condition without losing the developmental potency. In this study, AQP11 was examined instead. To compare expression pattern between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP11 by whole mount immunofluorescence. When the fertilized embryos were developed in the maternal tracts, the level of Aqp11 transcripts was decreased dramatically until 2-cell stage. Its level increased after 2-cell stage and peaked at 4-cell stage, but decreased again dramatically until morula stage. Its transcript level increased again at blastocyst stage. In contrast, the levels of Aqp11 transcript in embryos cultured in vitro were as follows. The patterns of expression were similar but the overall levels were low compared with those of embryos grown in the maternal tracts. AQP11 proteins were localized in submembrane cytoplasm of embryos collected from maternal reproductive tracts. The immune-reactive signals were detected in both trophectoderm and inner cell mass. However, its localization was altered in in vitro culture condition. It was localized mainly in the plasma membrane of the blastocysts contacting with external environment. The present study suggests that early stage embryo can develop successfully by themselves adapting to their environmental condition through modulation of the expression level and localization of specific genes like AQP11.

Regulation of AQP-4 Water Channel Expression in the Brain during Development and by Ischemia

  • Jung, Jin-Sup;Kim, Hae-Gyu;Bae, Hae-Rahn;Suh, Duk-Joon;Park, Hwan-Tae;Lee, Sang-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • 제1권5호
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    • pp.495-504
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    • 1997
  • Water transport is mediated by two distinct pathways, diffusional and channel-mediated water transport. The first molecular water channel was identified from human erythrocytes in 1992. Genetically-related proteins from other mammalian tissues have subsequently been identified to transport water, and the group is referred to as th "Aquaporins". Aquaporin-4 (AQP4) is most abundant in the brain, which may be involved in CSF reabsorption and osmoregulation. However, ontogeny and regulatory mechanisms of AQP4 channels have not been reported. Northern blot analysis showed that AQP4 mRNA began to be expressed in the brain just before birth and that its expression gradually increased by PN7 and then decreased at adult level. AQP4 was expressed predominantly in the ependymal cells of ventricles in newborn rats. And then its expression decreased in ependymal cells and increased gradually in other regions including supraoptic and paraventricular nuclei. AQP4 is also expressed in the subfornical organ, in which the expression level is not changed after birth. Cryogenic brain injury did not affect expression of AQP4 mRNA, while ischemic brain injury decreased it. Osmotic water permeability of AQP4 channel expressed in Xenopus oocytes was inhibited by the pretreatment of BAPTA/AM and calmidazolium, a $Ca^{2+}/Calmodulin$ kinase inhibitor, in a dose-dependent manner. These results indicate that the expression and the function of AQP4 channel are regulated by developmental processes and various pathophysiological conditions. These results will contribute to the understanding of fluid balance in the central nervous system and the osmoregulatory mechanisms of the body.

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Differential Expressions of Aquaporin Subtypes in the Adult Mouse Testis

  • Mohamed, Elsayed A.;Im, Ji Woo;Kim, Dong-Hwan;Bae, Hae-Rahn
    • 한국발생생물학회지:발생과생식
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    • 제26권2호
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    • pp.59-69
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    • 2022
  • Many efforts have been made to study the expression of aquaporins (AQP) in the mammalian reproductive system, but there are not enough data available regarding their localized expression to fully understand their specific roles in male reproduction. The present study investigated the expression and localization patterns of different AQP subtypes in the adult mouse testes and testicular spermatozoa using an immunofluorescence assay. All the studied AQPs were expressed in the testes and revealed subtype-specific patterns in the intensity and localization depending on the cell types of the testes. AQP7 was the most abundant and intensive AQP subtype in the seminiferous tubules, expressing in Leydig cells and Sertoli cells as well as all stages of germ cells, especially the spermatids and testicular spermatozoa. The expression pattern of AQP3 was similar to that of AQP7, but with higher expression in the basal and lower adluminal compartments rather than the upper adluminalcompartment. AQP8 expression was limited to the spermatogonia and Leydig cells whereas AQP9 expression was exclusive to tails of the testicular spermatozoa and elongated spermatids. Taken together, the abundance and distribution of the AQPs across the different cell types in the testes indicating to their relavance in spermatogenesis, as well as in sperm maturation, transition, and function.

생쥐 초기 배아에서 Aquaporin 8과 9의 발현에 관한 연구

  • 신현상;계명찬;강수만;이성은;이지원;강한승;김문규
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.68-68
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    • 2003
  • Aquaporin은 막관통 통로 단백질(transmembrane channel protein)로서, 삼투압의 농도구배에 따라 세포막을 가로질러 물분자를 이동시키는 기능을 하고 있다. 포유류 초기배아에서 포배강 형성은 영양외배엽세포에서 $Na^+ / K^+$ATPase에 의한 이온 농도 구배가 형성되면 auqaporin에 의해 물이 포배강으로 유입되면서 이루어진다. 본 연구에서는 생쥐 초기배아에서 반정량적인 역전사 중합효소 연쇄반응 방법(semi-quantitative RT-PCR)과 실시간 역전사 중합효소 연쇄반응 방법(real-time RT-PCR)을 통하여 AQP8과 9의 mRNA발현을 조사하고 다중 면역형광현미경 방법(confocal immunofluorescence microscopy)을 통해 단백질 발현양상을 분석하였다. AQP8 mRNA는 상실기까지 발현되지 않다가 포배기에 이르러 발현되었고 AQP9 mRNA는 수정란에서부터 발현되어 포배기에는 유의할 정도로 증가하였다. 따라서 AQP8 mRNA는 배아유전자가 활성화되어 나타나는 것이고 AQP9 mRNA는 모계유전자 기원임을 알 수 있었다. AQP8 단백질은 상실배 단계까지 발현되지 않다가 포배시기에 영양외배엽세포사이의 접합면에 발현되었고 AQP9 단백질은 상실배 시기에 할구 사이의 인접 부위에서 강하게 발현되었다가 포배시기에는 세포간의 접합면에 약하게 발현하는 경향을 나타내었다. 실시간 역전사 중합효소 연쇄반응 방법으로 조사한 결과 포배에서 물과 글리세롤을 통과시키는 AQP9는 mRNA의 발현양이 AQP8보다 약 4배 정도 많았다. 또한 포배기에 이르러서야 물만을 통과시키는 AQP8의 발현이 나타나는 것을 보아 포배강 형성시 외부에서 영양외배엽을 통해 포배강으로 유입되는 물의 이동(trans- trophectodermal water movements)에 AQP9보다 AQP8이 더 중요하게 관여할 것으로 사료된다., K, Pb, Cd, Cr, Co, Cu, Ni)을 측정하였다. 실험 조건1의 결과로서 각 국의 유아용 일회용 기저귀의 중금속 함량은 거의 유사한 경향을 나타내었으며 Cr, Zn, Pb, Ni, Mn, Mg, Li, K는 detection limit(2 ppm) 이하였고, Cd, Fe, Co, Cu, Ca, Al, Sr는 검출되었지만 기준치 이하였다. 실험 조건2의 결과로서 측정 항목(Cr, Sb, Cd, Pb, Ni, Co, Cu)중 Cr, Cd, Ni, Cu는 detection limit(0.1 ppm) 이하였고, Sb, Pb, Co는 검출되었지만 기준치 이하였다.았다. 4%의 경우에는 8$0^{\circ}C$이하로 온도를 낮추는 것이 좋은 상태를 나타내었다. 이와 같은 결과는 일반적으로 화학적 레팅을 4%, 7%에서한 선행결과와 상당히 다른 결과이다.염 농도가 증가할수록 감소 현상을 보였다.X>, 75BG30은 8.6$\mu\textrm{m}$, 75BG40은 7.02$\mu\textrm{m}$로 나타났다. 따라서 경화제 양에 관계없이 10$\mu\textrm{m}$ 이하로 나타나, 경화제 10$m\ell$만으로 미세한 크기를 얻을 수 있음을 알 수 있다. 젤리 강도 변화에 따른 차이는 300BF는 78.09$\mu\textrm{m}$ 300BG는 56.32$\mu\textrm{m}$로, 75BF나 75BG에 비하여 현저히 증가하여, 젤라틴의 젤리 강도는 캡슐 제조 조건의 주요한 변수임을 알 수 있다.추출물 투여시 혈당강하 및 혈중콜레스테롤 강하가 나타났으며, 상엽복합추출물 투여와 운동을 병행시 이러한 감소 효과가 더 뚜렷하게 나타났다.교육의 적임자로 보는 시각이 비교적 높았고 약 1/2정도는 영양교육에 참여하겠다는 의지를 가지고 있을 뿐만 아니라 실제로 영양지도를

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Molecular Analysis of AQP2 Promoter. I. cAMP-dependent Regulation of Mouse AQP2 Gene

  • Park, Mi-Young;Lee, Yong-Hwan;Bae, Hae-Rahn;Lee, Ryang-Hwa;Lee, Sang-Ho;Jung, Jin-Sup
    • The Korean Journal of Physiology and Pharmacology
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    • 제3권2호
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    • pp.157-164
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    • 1999
  • To determine molecular mechanisms of Aquaporin-CD (AQP2) gene regulation, the promoter region of the AQP2 gene was examined by transiently transfecting a promoter-luciferase reporter fusion gene into mouse renal collecting duct cell lines such as mIMCD-3, mIMCD-K2, and M-1 cells, and NIH3T3 mouse embryo fibroblast cells. PCR-Southern analysis reveals that mIMCD-3 and mIMCD-K2 cells express AQP2, but M-1 and NIH3T3 cells do not, and that the treatment with cpt-cAMP $(400\;{\mu}M)$) or forskolin/isobutylmethylxanthine (IBMX) increased the AQP2 expression in IMCD cells. In both IMCD and NIH3T3 cells, the constructs containing the promoter of AQP2 gene showed promoter activities, indicating lack of tissue-specific element in the 1.4 kb 5'-flanking region of the mouse AQP2 gene. Luciferase activity in the IMCD cells transfected with the construct containing 5-flanking region showed responsiveness to cpt-cAMP, indicating that the 1.4 kb 5'-flanking region contains the element necessary for the regulatory mechanism by cAMP. The promoter-luciferase constructs which do not have a cAMP-responsible element (CRE) still showed the cAMP responsiveness in IMCD cells, but not in NIH3T3 cells. Increase in medium osmolarity did not affect AQP2 promoter activity in mIMCD-K2 cells. These results demonstrate that AQP2 gene transcription is increased with cAMP treatment through multiple motifs including CRE in the 5'-flanking region of the gene in vitro, and the regulatory mechanism may be important for in vivo regulation of AQP2 expression.

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오가피(五加皮) 물추출물이 허혈-재관류로 유발된 급성 신부전에 미치는 영향 (Effect of Acanthopanacis cortex Water Extract on Renal Function in Ischemia/Reperfusion-lnduced Acute Renal Failure Rats)

  • 이안숙;강대길;김은주;양선녀;엄재연;안준석;이호섭
    • 동의생리병리학회지
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    • 제21권5호
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    • pp.1201-1209
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    • 2007
  • The present study was designed to examine whether water extract of Acanthopanacis cortex(AC) has an effect on renal functional parameters in association with the expression of aquaporin 2 (AQP-2) and heme oxygenase-1 (HO-1) in the ischemia/reperfusion induced acute renal failure (ARF) rats. Polyuria caused by down-regulation of renal AQP 2 in the ischemia-induced ARF rats was markedly restored by administration of AC (200 mg/kg, p.o.) with restoring expression of AQP 2 in the kidney. Administration of AC lowered the renal expression of HO-1, which was upregulated in rats with ischemia/reperfusion-induced ARF. The renal functional parameters including creatinine clearance, urinary sodium excretion, urinary osmolality, and solute-free reabsorption were also markedly restored in ischemia-ARF rats by administration of AC. Histological study also showed that renal damages in the ARF rats were abrogated by administration of AC. Taken together, the present data indicate that AC ameliorates renal defects in rats with ischemia/reperfusion-induced ARF.

급성 폐손상 동물모델에서 aquaporin 수분통로의 발현 (Aquaporin in bleomycin induced lung injury)

  • 장안수;박종숙;이준혁;박성우;김도진;어수택;김용훈;박춘식
    • Tuberculosis and Respiratory Diseases
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    • 제60권3호
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    • pp.330-336
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    • 2006
  • 배 경: 급성폐손상은 여러 가지 치료의 발전에도 높은 사망률을 보이는 질환으로 병태생리학적 연구가 필요한 실정이다. 아쿠아포린 (AQP)은 수분통로로 급성폐손상에서 폐부종에 관여되는 것으로 알려져 있어 bleomycin을 이용한 급성 폐손상모델에서 아쿠아포린의 발현을 확인하고자 하였다. 방 법: Bleomycin 10 U/kg을 Sprague-Dawley rats에 흡입시켜 급성 폐손상 보델을 만들고 흡입후 4, 7, 14일에 호흡수, 상시호흡량을 측정하고, 기관지폐포세척액을 얻으며, 폐조직을 얻었다. 얻은 폐조직에서 wet/dry 량을 측정하고 기관지 폐포세척액에서 사이토카인, 산화질소산물(nitrite와 nitrate)등을 측정하였다. 그리고 폐조직에서 AQP1, AQP4와 AQP5를 면역조직염색에서 확인하였다. 결 과: Bleomycin 흡입 후 호흡수는 4, 7일에 상시호흡량은 4일에 대조군에 비해 유의한 차이를 보였다. Wet/dry 질량은 대조군에 비해 4일에 유의하게 높았다. 기관지폐포세척액에서 산화질소산물은 4일에 가장 높았다. 면역화학염색에서 AQP1과 AQP4는 4일에 감소하였으며 AQP4는 7일에 증가하였고, AQP5는 4, 7, 14일에 발현이 증가하였다. 결 론: 아쿠아포린은 급성폐손상 모델에서 다양하게 발현하여 급성 폐손상의 경과에 따라 다르게 작용함을 시사한다.