• Title/Summary/Keyword: AQP6

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Differential Expressions of Aquaporin Subtypes in Female Reproductive Tract of Mice

  • Im, Ji Woo;Lee, Chae Young;Kim, Dong-Hwan;Bae, Hae-Rahn
    • Development and Reproduction
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    • v.24 no.3
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    • pp.177-185
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    • 2020
  • Although many aquaporin (AQP) transcripts have been demonstrated to express in the female reproductive tract, the defined localizations and functions of AQP subtype proteins remain unclear. In this study, we investigated the expression of AQP1, AQP3, AQP5, AQP6, and AQP9 proteins in female reproductive tract of mouse and characterized their precise localizations at the cellular and subcellular levels. Immunofluorescence analyses for AQP1, AQP3, AQP6, and AQP9 showed that these proteins were abundantly expressed in female reproductive tract and that intense immunoreactivities were observed in mucosa epithelial cells with a subtype-specific pattern. The most abundant aquaporin in both vagina and uterine cervix was AQP3. Each of AQP1, AQP3, AQP6, and AQP9 exhibited its distinct distribution in stratified squamous or columnar epithelial cells. AQP9 expression was predominant in oviduct and ovary. AQP1, AQP3, AQP6, and AQP9 proteins were mostly seen in apical membrane of ciliated epithelial cells of the oviduct as well as in both granulosa and theca cells of ovarian follicles. Most of AQP subtypes were also expressed in surface epithelial cells and glandular cells of endometrium in the uterus, but their expression levels were relatively lower than those observed in the vagina, uterine cervix, oviduct and ovary. This is the first study to investigate the expression and localization of 5 AQP subtype proteins simultaneously in female reproductive tract of mouse. Our results suggest that AQP subtypes work together to transport water and glycerol efficiently across the mucosa epithelia for lubrication, proliferation, energy metabolism and pH regulation in female reproductive tract.

Decreased Expression of Aquaporin-2 Water Channels in the Kidney in Rats Treated with Reserpine

  • Lee, Jong-Un;Oh, Yoon-Wha;Kim, Sun-Mi;Kim, Soo-Wan
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.4
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    • pp.307-313
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    • 2001
  • Whether there exists a sympathetic neural mechanism regulating the expression of aquaporin (AQP) water channels in the kidney was investigated. Male Sprague-Dawley rats were treated with reserpine (1 mg/kg, IP), and the expression of AQP1-4 proteins was determined in the kidney one day thereafter. Following the treatment with reserpine, the systolic blood pressure measured in a conscious state was significantly decreased in the experimental group compared with that in the control $(83{\pm}8\;vs\;124{\pm}6\;mmHg;\;n=6\;each,\;P<0.05)$. The expression of AQP2 proteins was decreased in the cortex, outer medulla, and inner medulla. The decrease of AQP2 proteins was in parallel in the membrane and the cytoplasmic fractions, suggesting a preserved AQP2 targeting. No significant changes were observed in the expression of AQP1, AQP3, or AQP4. Neither basal nor AVP-stimulated formation of cAMP was significantly altered. These results suggest that the sympathetic nervous system has a tonic stimulatory effect specifically on the expression of AQP2 water channels in the kidney.

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Induction of anti-aquaporin 5 autoantibodies by molecular mimicry in mice

  • Lee, Ahreum;Choi, Youngnim
    • International Journal of Oral Biology
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    • v.45 no.4
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    • pp.211-217
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    • 2020
  • Molecular mimicry is the most common mechanism that breaches self-tolerance. We previously identified autoantibodies to aquaporin-5 (AQP5) in the sera of patients with Sjögren's syndrome and found that the aquaporin of Prevotella melaninogenica (PmAqp), an oral commensal, is highly homologous to human AQP5. This study aimed to test whether PmAqp can induce anti-AQP5 autoantibodies via molecular mimicry. From the amino acid sequence of PmAqp, an immunizing peptide; i.e., PmE-L, was designed, which contained both the B cell epitope "E" and T cell epitope. C57BL/6 and BALB/c mice were subcutaneously immunized with linear or cyclic forms of PmE-L emulsified in incomplete Freund's adjuvant. The concentrations of the antibodies in sera were measured using enzyme-linked immunosorbent assays. Both linear and cyclic PmE-L induced high levels of antibodies against not only the immunized peptides but also autoantibodies against AQP5E and antibodies against PmE, a Pm homolog of AQP5E. In C57BL/6 mice; however, the cyclic form of PmE-L was more efficient than the linear form in inducing autoantibodies against AQP5E that contained a cyclic epitope. The levels of anti-PmE antibodies and anti-AQP5E autoantibodies showed a strong positive correlation (r = 0.95, p < 0.0005), suggesting molecular mimicry. Collectively, the mice produced anti-AQP5E autoantibodies in response to a PmAqp-derived peptide. This model proved to be useful for studying the mechanisms of autoantibody production by molecular mimicry.

Aquaporin-3 Downregulation in Vitiligo Keratinocytes Increases Oxidative Stress of Melanocytes

  • Nan-Hyung Kim;Ha Jung Kim ;Ai-Young Lee
    • Biomolecules & Therapeutics
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    • v.31 no.6
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    • pp.648-654
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    • 2023
  • Oxidative stress-induced melanocyte apoptosis is linked to the immune system and plays a critical role in the pathogenesis of vitiligo. Aquaporin-3 (AQP3), which is downregulated in vitiligo keratinocytes, regulates intracellular H2O2 accumulation. However, the role of AQP3 in oxidative stress is uncertain in vitiligo. This study investigated the effect of downregulated AQP3 on oxidative stress in vitiligo using lesional and non-lesional skin specimen sets from vitiligo patients and primary cultured adult normal human epidermal keratinocytes, with or without downregulation and overexpression of AQP3 in the presence or absence of H2O2 treatment. The levels of nuclear factor E2-related factor 2 (NRF2) and/or its main target, NAD(P)H quinone dehydrogenase 1 (NQO-1), were lower in the lesional keratinocytes and cultured keratinocytes with AQP3 knockdown, but were increased in keratinocytes upon AQP3 overexpression. Ratios of NRF2 nuclear translocation and NQO-1 expression levels were further reduced in AQP3-knockdown keratinocytes following H2O2 treatment. The conditioned media from AQP3-knockdown keratinocytes treated with H2O2 contained higher concentrations of reactive oxygen species (ROS). Moreover, the number of viable melanocytes was reduced when the conditioned media were added to the culture media. Overall, AQP3 downregulation in the keratinocytes of patients with vitiligo can induce oxidative stress in neighboring melanocytes, leading to melanocyte death.

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.

Ethanol Extract of Astragalus membranaceus Bunge Induces Aquaporin-3 Expression in HaCaT Cells (HaCaT Cell에서 황기 에탄올 추출물의 Aquaporin-3 발현 효과)

  • Park, Hyun-Chul;Kim, Hee-Taek;Ha, Hun-Young;Lee, Pyeong-Jae;Yoon, Kyung-Sup
    • KSBB Journal
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    • v.28 no.6
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    • pp.394-399
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    • 2013
  • Astragalus membranaceus Bunge is used in herbal medicine in Eastern Asian countries including Korea. In this study, we assessed the effects of A. membranaceus extract (AM) on the aquaporin-3 (AQP3) protein expression in HaCaT cells. AM did not affect viability of HaCaT cells. AQP3 expression and cell migration seem to be maximal at $100{\mu}g/mL$ concentration. Epidermal growth factor receptor (EGFR) kinase inhibitor, PD153035, blocked AM-induced AQP3 expression and cell migration. In addition, an 80% ethanol extracts of herbal prescription, SinhyoTakleesan (ST), which is composed of A. membranaceus, Angelicae gigantis, Glycyrrhiza glabra Linne, and Lonicera japonica Flos also induced AQP3 expression at $20{\mu}g/mL$ in HaCaT cells. Collectively, these results suggest that AM induce AQP3 expression via EGFR pathway.

Characterization and expression profiles of aquaporins (AQPs) 1a and 3a in mud loach Misgurnus mizolepis after experimental challenges

  • Lee, Sang Yoon;Nam, Yoon Kwon;Kim, Yi Kyung
    • Fisheries and Aquatic Sciences
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    • v.20 no.9
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    • pp.23.1-23.9
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    • 2017
  • Two distinct cDNAs encoding aquaporins (mmAQPs 1a and 3a) were isolated and characterized from mud loach Misgurnus mizolepis. The identified mud loach AQP cDNAs encode for polypeptides of 260 and 302 amino acids. Topology predictions confirmed six putative membrane-spanning domains connected by five loops and the N- and C-terminal domains being cytoplasmic. The mud loach AQPs 1a and 3a showed broad distribution in multiple tissues including immune-responsive tissues as well as osmoregulatory tissues. Hence, the diversity of AQP distribution and expression possibly indicated its differential functions in the regulation of fluid movement in response to environmental stimuli. The transcription of mmAQP genes was differentially modulated by immune challenges. In particular, the mmAQP3a expression level in the liver was more responsive to immune challenges than that of mmAQP1a. Taken together, fish stimulation or infection resulted in significant modulation of mud loach AQP genes, suggesting potential functional roles of these proteins in piscine pathophysiological process.

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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Induction of Anti-Aquaporin 5 Autoantibody Production by Immunization with a Peptide Derived from the Aquaporin of Prevotella melaninogenica Leads to Reduced Salivary Flow in Mice

  • Ahreum Lee;Duck Kyun Yoo;Yonghee Lee;Sumin Jeon;Suhan Jung;Jinsung Noh;Soyeon Ju;Siwon Hwang;Hong Hee Kim;Sunghoon Kwon;Junho Chung;Youngnim Choi
    • IMMUNE NETWORK
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    • v.21 no.5
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    • pp.34.1-34.16
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    • 2021
  • Sjögren's syndrome (SS) is an autoimmune disease characterized by dryness of the mouth and eyes. The glandular dysfunction in SS involves not only T cell-mediated destruction of the glands but also autoantibodies against the type 3 muscarinic acetylcholine receptor or aquaporin 5 (AQP5) that interfere with the secretion process. Studies on the breakage of tolerance and induction of autoantibodies to these autoantigens could benefit SS patients. To break tolerance, we utilized a PmE-L peptide derived from the AQP5-homologous aquaporin of Prevotella melaninogenica (PmAqp) that contained both a B cell "E" epitope and a T cell epitope. Repeated subcutaneous immunization of C57BL/6 mice with the PmE-L peptide efficiently induced the production of Abs against the "E" epitope of mouse/human AQP5 (AQP5E), and we aimed to characterize the antigen specificity, the sequences of AQP5E-specific B cell receptors, and salivary gland phenotypes of these mice. Sera containing anti-AQP5E IgG not only stained mouse Aqp5 expressed in the submandibular glands but also detected PmApq and PmE-L by immunoblotting, suggesting molecular mimicry. Characterization of the AQP5E-specific autoantibodies selected from the screening of phage display Ab libraries and mapping of the B cell receptor repertoires revealed that the AQP5E-specific B cells acquired the ability to bind to the Ag through cumulative somatic hypermutation. Importantly, animals with anti-AQP5E Abs had decreased salivary flow rates without immune cell infiltration into the salivary glands. This model will be useful for investigating the role of anti-AQP5 autoantibodies in glandular dysfunction in SS and testing new therapeutics targeting autoantibody production.

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.