• Title/Summary/Keyword: AQP2

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생쥐 초기 배아에서 Aquaporin 8과 9의 발현에 관한 연구

  • 신현상;계명찬;강수만;이성은;이지원;강한승;김문규
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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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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Effects of Recombinant Aquaporin 3 and Seawater Acclimation on the Expression of Aquaporin 3 and 8 mRNAs in the Parr and Smolt Stages of Rainbow Trout, Oncorhynchus mykiss

  • Kim, Na Na;Choi, Young Jae;Lim, Sang-Gu;Kim, Bong-Seok;Choi, Cheol Young
    • Ocean and Polar Research
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    • v.38 no.2
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    • pp.103-113
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    • 2016
  • This study aimed to examine the role of two aquaporin isoforms (AQP3 and AQP8) in response to the hyperosmotic challenge of transitioning from freshwater (FW) to seawater (SW) during parr and smoltification (smolt) using the rainbow trout, Oncorhynchus mykiss. We examined the changes in the expression of AQPs mRNAs in the gills and intestine of the parr and smolt stages of rainbow trout transferred from FW to SW using quantitative real-time PCR in an osmotically changing environment [FW, SW, and recombinant AQP3 (rAQP3) injection at two dosage rates]. Correspondingly, AQPs were greater during smoltification than during parr stages in the rainbow trout. Plasma osmolality and gill $Na^+/K^+$-ATPase activity increased when the fish were exposed to SW, but these parameters decreased when the fish were exposed to SW following treatment with rAQP3 during the transition to seawater. Our results suggest that AQPs play an important role in water absorbing mechanisms associated with multiple AQP isoforms in a hyperosmotic environment.

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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    • v.1 no.5
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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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Adaptive Transition of Aquaporin 5 Expression and Localization during Preimplantation Embryo Development by In Vitro Culture

  • Park, Jae-Won;Shin, Yun Kyung;Choen, Yong-Pil
    • Development and Reproduction
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    • v.18 no.3
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    • pp.153-160
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    • 2014
  • Adaptive development of early stage embryo is well established and recently it is explored that the mammalian embryos also have adaptive ability to the stressful environment. However, the mechanisms are largely unknown. In this study, to evaluate the possible role of aquaporin in early embryo developmental adaptation, the expression of aquaporin (AQP) 5 gene which is detected during early development were examined by the environmental condition. To compare expression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5 by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantation embryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expression pattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second mid-preimplantation gene activation phase, but the fold changes were modified between in vivo embryos and in vitro embryos. During in vivo development, AQP5 was mainly localized in apical membrane of blastomeres of 4-cell and 8-cell stage embryos, and then it was localized in cytoplasm. However, the main localization area of AQP5 was dramatically shifted after 8-cell stage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

Validity and Reliability of Adversity Quotient Profile for Measuring Overcoming of Adversity among Nurses in Korea (역경지수(Adversity Quotient Profile) 도구의 타당성 및 신뢰성 검증 -종합병원 간호사 중심으로-)

  • An, Ji-Yeon;Woo, Hae-Young;Song, Jung-Hee;Kim, Hye-Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.4
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    • pp.2285-2294
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    • 2014
  • Stoltz's AQP is a tool to estimate the level of adversity. The purpose of study is to test the reliability and validity of AQP. The participants were 297 nurses of tertiary hospital in Korea. Data were collected from Aug. to Oct. 2013. Resilience Scale was used as criterion of AQP. Cronbach's alpha test, item total correlation, exploratory factor, correlation analysis, and ANOVA test were used. In result, factor analysis was accounted for four factors explaining 56.256% of total variance and corresponding factors were factor 1 (Control, 10.7%), factor 2 (Ownership, 13.7%), factor 3(Reach, 17.4%), factor 4 (Endurance, 14.9%). The internal consistency was acceptable (Cronbach's alpha=.84). Four factors were positively correlated with RS. AQP is a reliable and valid instrument to measure for adversity quotient in Korean nurse.

Expression of Aquaporin 1 in Bladder Uroepithelial Cell Carcinoma and its Relevance to Recurrence

  • Liu, Jie;Zhang, Wei-Yi;Ding, De-Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3973-3976
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    • 2015
  • Objectives: To explore the expression of aquaporin 1 ($AQP_1$) in bladder uroepithelium cell carcinoma (BUCC) and its relevance to recurrence. Materials and Methods: Tissue samples from 45 BUCC patients who underwent total cystectomy or transurethral resection of bladder tumor (TURBT) and from 40 patients with non-bladder cancers who underwent special detection or treatments were collected. The level of expression of $AQP_1$ in BUCC tissues and normal bladder tissues was assessed by immunohistochemistry so as to analyze the relevance to pathological patterns and time of recurrence in BUCC patients. Results: The expression levels of $AQP_1$ normal bladder tissues and BUCC tissues were $2.175{\pm}0.693$ and $3.689{\pm}0.701$, respectively, and the difference was significant (t=9.99, P<0.0001). Marked increase was noted with BUCC histological grade and pathological stage (P<0.01). Moreover, the expression of $AQP_1$ was evidently higher in cancerous tissues with lymph node metastasis than in those without (P<0.01). With short-term recurrence, the positive cell expression rate of $AQP_1$ was higher in primary tissues, which increased obviously after recurrence. Additionally, the recurrent time of BUCC was negatively associated with the positive cell expression rate of $AQP_1$ and the difference between the expression of $AQP_1$ before and after recurrence (r=-0.843, F=39.302, P=0.000; r=-0.829, F=35.191, P=0.000). Conclusions: $AQP_1$, which reflects the grade, stage, lymph node metastasis and recurrence of BUCC, has potential guiding significance in the diagnosis and treatment of bladder cancarcinoma.

Temporal Aquaporin 11 Expression and Localization during Preimplantation Embryo Development

  • Park, Jae-Won;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.19 no.1
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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.

Effect of Steroid Hormones on Expression and Localization of Aquaporin-4, -5 and -8 Genes in Mouse Uterine Endometrium (스테로이드 호르몬이 생쥐 자궁내막에서 Aquaporin-4, -5와 -8 유전자의 발현과 존재부위에 미치는 영향)

  • Kang, Soo-Man;Kang, Han-Seung;Gye, Myung-Chan;Shin, Hyeon-Sang;Lee, Ji-Won;Lee, Sung-Eun;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.2
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    • pp.119-131
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    • 2004
  • 연구 목적: 난소에서 분비되는 스테로이드 호르몬인 에스트로젠과 프로게스테론은 포유동물의 생식기관 발달과 정상적인 생식 기능, 수정과 배아의 착상에 중요한 역할을 한다. 특히 에스트로젠은 자궁내액을 내강으로 분비하여 자궁부종 기작에 중요한 역할을 한다. 자궁내액은 정자의 수정능력 획득과 착상전 배아의 발달에 매우 중요하다. Aquaporin (AQP)은 막관통 물수송 단백질로서 여러 조직에 넓게 분포되어 있으며, 세포간 또는 상피세포간 물의 이동에 중요한 역할을 한다. 본 연구에서는 생쥐 자궁에서 스테로이드 호르몬에 의해 조절되는 자궁내액의 이동에 AQP 유전자가 관여하는지를 알아보았다. 연구 재료 및 방법: 난소 절제술을 시행한 생쥐에 스테로이드 호르몬을 피하주사하고 6, 12, 24시간 간격으로 자궁조직을 적출하였다. 대조군은 sesame oil만을 주사한 후 6시간째에 수획한 자궁조직을 사용하였으며, 실험군은 시간대별과 스테로이드 처리별로 채취한 자궁조직에서 역전사중합효소반응을 수행하였다. 역전사중합효소반응을 통해 막관통 단백질인 AQP-4, -5, -8 mRNA의 발현양상을 살펴보았다. 또한 mRNA의 위치를 살펴보기 위해 laser microdissection을 이용하여 RT-PCR을 수행하였다. 마지막으로 자궁조직내에서의 단백질 발현 부위를 관찰하기 위해 면역조직화학염색을 실시하였다. 결 과: AQP-4, -5, -8 mRNA은 프로게스테론을 처리한 군보다 에스트로젠을 처리한 군에서 많이 발현되었으며, 에스트로젠을 주사한 지 6시간째 발현정도를 대조군과 비교할 때 AQP-4, -5, -8 mRNA가 각각 7.9배, 2.8배, 3.8배로 나타났다. AQP-4, -5, -8 mRNA는 간충조직보다 자궁내 상피조직에서 스테로이드 호르몬의 영향을 받아 발현양상의 차이가 나타났으며, 주로 에스트로젠의 영향을 받아 발현이 증가하였다. AQP-4 단백질은 에스트로젠을 24시간 처리한 후 프로게스테론을 처리한 군의 자궁내 상피조직에서 많이 발현되었으며, AQP-5와 -8 단백질은 에스트로젠을 처리한 군의 자궁내 상피조직에서 발현이 증가하였다. 결 론: 이상의 결과를 통해 AQP-4, -5, -8은 주로 에스트로젠에 의해 자궁내 상피세포에서 발현이 증가되는 것으로 보아 에스트로젠의 영향하에 일어나는 자궁내액의 이동으로 인한 자궁부종기작에 이동통로로서 관여하는 것으로 사료된다.

Effects of Boheotang-gagam on Milk Production and Factors Related Lactation in Postpartum Mice (보허탕가감 투여가 산후 생쥐의 유즙분비량 및 유즙분비 관련 인자에 미치는 영향)

  • Lee, Ah-Yeong;Lee, Eun-Hee;Im, Ji-Yeong;Kim, Hong-Jun;Lee, Chang-Hyun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.29 no.1
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    • pp.35-52
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    • 2016
  • Objectives :The purpose of this study was to investigate the effect on factors related the expression of aquaporins (AQP) and milk production after administration of Boheotang-gagam in lactating mice. Methods: The SKH-1 mice were randomly allocated to the control group which was administered with distilled water for two weeks after the parturition and the experimental groups such as, lactating+400G group (L400G) which was administered with Boheotang-gagam 400 mg/day, lactating+600G group (L600G) which was administered with 600 mg/day for two weeks after the parturition, and 400G+lactating+400G group (400G-L400G) which was administered with 400 mg/day for 3 weeks starting one week prior to parturition for experiment (n=6 per group). Results: 1. With regard to the immunohistochemical staining reaction for AQP1, AQP3, and AQP5, stronger immune response was also showed in mammary gland in all experimental groups as compared to the control group. AQP1 showed stronger immune response in the capillaries and venules which were located around the interlobular duct, while stronger immune response of AQP3 and AQP5 showed in the secretory alveolar epithelia and intralobular and interlobular ductal epithelial cells. 2. In the western blot, L400G group showed the most increased expression followed by L600G and then 400G-L400G group in AQP1. In AQP3, the order of expression density was observed as L600G, 400G-L400G and L400G group. Lastly, in AQP5, L400G group presented the most increased expression followed by L600G and 400G-L400G group. Conclusions: Boheotang-gagam would have the effect of increasing the lactation of mice after the birth by increasing the prolactin level and adjusting the expression of AQPs and prolactin receptor in the mammary glands.

Effects of Bojungchiseup-tang on Renal Expression of Water Channels, Na, K-ATPase and Nitric Oxide Synthase in Rats (보중치습탕의 백서 신장 수분채널, Na, K-ATPase, 산화질소 합성효소 발현에 미치는 영향)

  • Kang Dae Gill;Kim Jang Giun;Kim Bok Hae;Cho Dong Ki;Sohn Eun Jin;Ryu Do Gon;Lee Ho Sub
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.1
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    • pp.72-77
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    • 2002
  • The present study was examined the effects of Bojungchiseup-tang water extract on the renal expression of renal function regulatory proteins including aquaporin 2 (AQP 2), aquaporin 3 (AQP 3), Na, K-ATPase α1 subunit, endothelial nitric oxide synthase (ecNOS), and inducible nitric oxide synthase (iNOS) in rats. The renal expression of AQP 3 was attenuated in rats administered with Bojungchiseup-tang water extract without altered expression of AQP 2, while ecNOS was up-regualted. Oral administration of Bojungchiseup-tang water extract (40 ㎕/100 g) also attenuated the renal expression of Na, K-ATPase α1-subunit and iNOS protein. These results suggest that the diuretic and natriuretic effects of Bojungchiseup-tang maybe causely related with a decreased expression of AQP 3 and increased expression of ecNOS.