Molecular Cloning and Expression of a cDNA Encoding the Aquaporin Homologue from the Firefly, Pyrocoelia rufa

Lee, Kwang-Sik,Kim, Seong-Ryul,Lee, Sang-Mong,Lee, Kyung-Ro,Sohn, Hung-Dae,JIn, Byung-Rae 한국곤충학회 2001 Entomological Research Vol.31 No.4

Aquaporin유전자 계열에 속하는 한 유전자를 결정하는 cDNA를 늦반딧불이 Pyrocoelia rufa의 cDNA library로부터 클로닝하였다. 늦반딧불이 aquaporin cDNA의 염기서열 분석 결과, 813bp의 길이로 271개의 아미노산으로 구성된다. 늦반딧불이 aquaporin의 단백질 서열은 이미 알려진 곤충 유래 aquaporin 및 일부 포유동물 유래 aquaporin들과 상동성을 나타내었다. 늦반딧불이 aquaporin의 단백질 서열 역시 aquaporin계열에서 특징적으로 보존된 NPA motifs가 두 영역에서 관찰되었다. 또한 늦반딧불이 aquaporin과 이미 알려진 aquaporin계열의 단백질 서열을 이용한 계통 분석 결과, 늦반딧불이 aquaporin은 이미 알려진 곤충 유래 aquaporin계열에 속하였다. 전사수준에서 늦반딧불이 aquaporin유전자의 발현을 알아보기 위하여, 세가지 특정적인 조직 (지방체, 중장 및 피부)으로부터의 mRNA를 이용하여 Northern blot분석을 수행하였다. 그 결과 늦반딧불이 aquaporin유전자는 대부분의 조직에서 발현되는 것으로 추정된다. 한편, 늦반딧불이의 aquaporin유전자를 곤충 베클로바이러스 발현 벡터계를 이용하여 곤충 세포주에서 발현한 결과 34kDa의 밴드로 관찰되었다. A cDNA encoding a putative member of the aquaporin gene family was cloned from a cDNA library of the firefly, pyrocoelia rufa. Sequence analysis of the cDNA encoding the aquaporin homologue of P. rufa revealed that the 813 bp cDNA has an open reading frame of 271 amino acid residues. The deduced protein sequence of the aquaporin homologue gene of P. rufa was aligned to the insect aquaporins and several mammalian aquaporins. The protein sequence of p. rufa aquaporin homologue reveals two regions of NPA motifs conserved in the aquaporin family. Phylogenetic analysis further confirmed the deduced protein sequence of the p. rufa aquaporin homologue gene to be belonged to the insect aquaporin family. Northern blot analysis suggested expression of the p. rufa aquaporin homologue gene in most, if not all, body tissues at the transcriptional level. The cDNA encoding the aquaporin homologue of P. rufa was expressed as a 34 kDa band in baculovirus-infected insect cells.

장기 복막투석 동물 모델에서 물수송체의 역할

박민선(MS Park),차정호(JH Cha),김진(J Kim),(Soren Nielsen) 대한신장학회 2000 Kidney Research and Clinical Practice Vol.19 No.2

N/A Sufficient fluid removal is vital to renal replace-ment therapy in end-stage renal failure patients. Aquaporins are integral membrane proteins and the primary water channels that allow water transport only. Type 1, 3 and 4 aquaporins were found in peritoneal capillary walls and peritoneal mesothelial cells. Approximately 5096 of total amount of free water transported during peritoneal dialysis is through aquaporins. Ultrafiltration failure and fluid overload are found in some of long-term continous ambulatory peritoneal dialysis(CAPI3) patients and are major causes of withdrawl from CAPD. Long- term use of high glucose containing dialysis solutions, and functional and morphological changes of aquaporins were suggested as possible mechanisms of ultrafiltration failure. However, a direct relation between alterations of aquaporins in the peritoneum and ultrafiltration failure in long-term CAPD has not been reported yet. In this study peritoneal aquaporins and ultrafiltration were evaluated after long-term peritoneal exposure to high glucose containing dialy- sis solutions in rats. Sprague-Dawley rats with normal kidney func- tions were used. Twenty five milliliter of 4.25% glucose containing dialysis solutions were injected into the peritoneal cavity twice a day for 12 weeks in 13 rats(dialysis-group). The other 13 rats were used without intraperitoneal injection as controls (control-group). One rat from each group died during the study was excluded. After 12 weeks of intraperitoneal injection, a 2 hour peritoneal transport study was done in 9 rats from each group. To calculate intraperitoneal fluid absorption rate, (131)I labelled human serum albumin(RISA) was used as a volume marker. Mesenteries were taken from the remaining three rats from each group for immuno-histochemistry for aquaporin type l. Intraperitoneal volume after 2 hour dialysis was significantly lower in dialysis-group than in control-group(33.7±3.6 vs 39.4±6.1mL, p<0,05). The peri- toneal fluid absorption rate was significamtly higher in dialysis-group than in contml-group(0,070±0.051 vs 0.049±0.016 mL/min, p<0.05). Dg'P> srxlium was signifieantly higher in dialysis-group than in con- trol-group(0.890±0.014 vs 0.856±0.038, p<0.05).D₂P₂urea and D2/D0 glucose did not differ between the two groups. Immunohistochemistry revealed that aquaporin type 1 was strongly stained in the me-sentery capillary walls in control-group, while it was almost disappeared in dialysis-group. In conclusion, long-term use of high glucose containing dialysis solutions decreased aquaporin type 1 population in the peritoneum and ultrafil-tration volume. Increased peritoneal fluid absorption rate is also in part responsible for decreased ultra- filtration volume after long-term use of dialysie solutions.

The role of aquaporin-5 in cancer cell migration: A potential active participant

Jensen, H.H.,Login, F.H.,Koffman, J.S.,Kwon, T.H.,Nejsum, L.N. Pergamon 2016 The international journal of biochemistry & cell b Vol.79 No.-

Emerging data identifies the water channel aquaporin-5 as a major player in multiple cancers. Over-expression of aquaporin-5 has been associated with increased metastasis and poor prognosis, suggesting that aquaporin-5 may enhance cancer cell migration. This review aims to highlight the current knowledge and hypothesis regarding downstream signaling partners of aquaporin-5 in relation to cancer cell migration. The molecular mechanisms that link aquaporin-5 to cell migration are not completely understood. Aquaporin-5 may promote cell movement by increasing water uptake into the front of the cell allowing local swelling. Aquaporin-5 may also activate extracellular-regulated kinases, increasing proliferation and potentially stimulating the migration machinery. Thus, further studies are warranted to identify the underlying mechanisms and signaling pathways. This will reveal whether aquaporin-5 and downstream effectors could be targets for developing new cancer therapeutics.

An electrokinetic approach to fabricating aquaporin biomimetic membranes for water purification

Fuwad, Ahmed,Ryu, Hyunil,Lee, Jun-Hee,Kim, Daejoong,Yoo, Yeong-Eun,Kim, Young-Rok,Kim, Sun Min,Jeon, Tae-Joon Elsevier 2019 Desalination Vol.452 No.-

<P><B>Abstract</B></P> <P>Membrane technology has been dominating the water desalination industry for decades due to its high efficiency and reliability. However, conventional membrane materials present performance limitations; thus, the demand for the development of new material is high. In recent years, aquaporin biomimetic membranes have emerged as a next-generation water desalination platform based on natural phenomena. Aquaporin is a natural water-selective protein that possesses exceptional water selectively and permeability properties. However, aquaporin must be embedded in an amphiphilic structure, such as a cell membrane, and the mimetic structure and stability of these environments represent key factors for successful water purification systems. Herein, we report an electrokinetic approach that stabilizes the aquaporin-containing membranes on a porous substrate under an applied electric field, resulting in an exceptionally stable and uniform biomimetic membrane on a solid support. The surface morphological analysis shows that the liposomes retained their perfect shape and size and did not present fusion or aggregation. Moreover, under forward osmosis, our membrane presents a salt rejection rate that reached 97.8±0.7% with 7.45±0.62Lm<SUP>−2</SUP>h<SUP>−1</SUP> (LMH) of water flux.</P> <P><B>Highlights</B></P> <P> <UL> <LI> An electrokinetic approach that stabilizes aquaporin biomimetic membranes on a porous substrate is demonstrated in this work. </LI> <LI> Charged proteoliposomes are concentrated on the pores under an applied electric field </LI> <LI> Forward osmosis was applied to the aquaporin biomimetic membranes, resulting in high salt rejection </LI> <LI> An electrokinetic method can add great versatility to water purification systems using aquaporin-functionalized biomimetic membranes </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Genome-wide analysis and expression profiling of the Solanum tuberosum aquaporins

Venkatesh, J.,Yu, J.W.,Park, S.W. Gauthier-Villars ; Elsevier Science Ltd 2013 Plant physiology and biochemistry Vol.73 No.-

Aquaporins belongs to the major intrinsic proteins involved in the transcellular membrane transport of water and other small solutes. A comprehensive genome-wide search for the homologues of Solanum tuberosum major intrinsic protein (MIP) revealed 41 full-length potato aquaporin genes. All potato aquaporins are grouped into five subfamilies; plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), NOD26-like intrinsic proteins (NIPs), small basic intrinsic proteins (SIPs) and x-intrinsic proteins (XIPs). Functional predictions based on the aromatic/arginine (ar/R) selectivity filters and Froger's positions showed a remarkable difference in substrate transport specificity among subfamilies. The expression pattern of potato aquaporins, examined by qPCR analysis, showed distinct expression profiles in various organs and tuber developmental stages. Furthermore, qPCR analysis of potato plantlets, subjected to various abiotic stresses revealed the marked effect of stresses on expression levels of aquaporins. Taken together, the expression profiles of aquaporins imply that aquaporins play important roles in plant growth and development, in addition to maintaining water homeostasis in response to environmental stresses.

사람과 흰쥐의 고환에서 Aquaporin 유전자의 발현

박남철,박영수,오금수,정진섭,Park, Nam-Cheol,Park, Young-Soo,Oh, Gom-Su,Jung, Jin-Sup 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.2

Objective: Several water channels (aquaporins; AQP) that belong to the MIP (major intrinsic protein) family have identified. In the selected tissues including red blood cells or renal tubules, water movements are abundant and/or physiologically important. Unexpectedly, a high water permeability of human and ram sperm has been reported. Recent studies showed that AQP7 and AQP8 are present in testes, so that the high water permeability of human sperm suggested to be mediated by AQPs. Method: To identify the identity of aquaporins expressed in testes, RT-PCR was performed using degenerative primers, which were designed to correspond to highly conserved sequences surrounding the Asn-Pro-Ala (NPA) motifs in the aquaporins. New expressed AQP series were reconfirmed by immunohistochemical study using rabbit polyclonal antibodies. Results: DNA sequencing of PCR products revealed that AQP2 and AQP3 mRNA as well as AQP7 and AQP8 are expressed in human and rat testes. In human and rat testes, AQP2 are expressed in spermatozoa, interstitial cells and myofibroblasts and AQP3 are expressed in myofibroblasts of semineferous tubules on immunocytochemical stain. Conclusion: These results indicate that multiple aquaporins are expressed in testes, and that they may have important roles in the spermatogenesis and the germ cell function of testis.

전자간증 임부의 태반에서 Aquaporin-8의 발현

하민숙 ( Min Sook Ha ),황태기 ( Tae Gi Hwang ),이상경 ( Sang Kyeong Lee ),홍승화 ( Seung Hwa Hong ),김학순 ( Hak Soon Kim ),박연진 ( Yeon Jin Park ),이인하 ( In Ha Lee ),정은환 ( Eun Hwan Jeong ) 대한산부인과학회 2004 Obstetrics & Gynecology Science Vol.47 No.12

목적 : 세포내외의 수분통로 역할을 하는 aquaporin-8 유전자가 사람 태반에서 발현되는지 확인하고 발현된다면 중증 전자간증 임부와 정상 임부간에 발현 정도의 차이가 있는지 알아보고자 하였다. 연구 방법 : 충북대학교병원에서 분만 진통이 없이 제왕절개술을 시행하여 분만한 중증 전자간증 임부와 정상 임부의 태반에서 aquaporin-8 cRNA probe를 이용하여 in situ hybridization을 시행하고 광학 현미경으로 aquaporin- Objective : This study was to determine whether aquaporin-8, which plays a role as a transcellular water channel, is expressed in human placenta, and to compare the degree of its expression between preeclamptic women and normal pregnant women. Methods : P

An herbal medicine prescription (Oreongsan) developed as a new alternative treatment in patients with chronic subdural hematoma: a narrative review

권승원,Chul Jin,조기호 한국한의학연구원 2019 Integrative Medicine Research Vol.8 No.1

An herbal medicine prescription, Oreongsan (ORS), which is composed of Polyporus, Alismatis Rhizoma, Atractylodis Rhizoma, Poria (Hoelen), and Cinnamomi Cortex Spissus, has been used as treatment in patients with various symptoms such as thirst, diminished urination, edema, hangover, and diarrhea. ORS is the representative prescription of the ’inducing diuresis’ (isu) effect, which traditionally means the effect of controlling the water balance. Advancement of modern science has enabled the determination of the action mechanism of herbal medicine complexes. As a result, ORS has been used in the treatment of patients with chronic subdural hematoma (CSDH), representing a novel indication. ORS inhibits the upregulation of aquaporin-4, which is involved in the development of brain edema in the central nervous system. Both aquaporin-1 and aquaporin-4 are expressed in the outer membrane of the CSDH; through its effect as aquaporin-4 inhibitor, ORS prevents the inflow of fluid into the hematoma, thereby preventing the development and recurrence of hematoma. In this study, we reviewed the relationship between the inducing diuresis effect of ORS and aquaporin, conservative treatment approach in patients with CSDH, and the prevention of recurrence in patients undergoing combined burr hole surgery and treatment with ORS.

흰 쥐에서 양측 요관 폐쇄 해제 후 신장의 Aquaporin-2와 Na-K-2Cl Cotransporter의 발현 변화 및 항이뇨호르몬 치료의 효과

신진호 ( Jin Ho Shin ),성수아 ( Su Ah Sung ),서지아 ( Ji A Seo ),한금현 ( Kum Hyun Han ),조원용 ( Won Yong Cho ),표희정 ( Heui Jung Pyo ),유기환 ( Kee Hwan Yoo ),원남희 ( Nam Hee Won ) 대한신장학회 2003 Kidney Research and Clinical Practice Vol.22 No.1

목적 : 혈액 용적 감소에 관련된 신장의 aquaporin-2 (AQP2) 수분 통로 발현 변화를 용적 감소 수 경과 시간에 따라 조사하였다. 방법 : Sprague-Dawley 숫쥐에서 몸무게의 약 2%에 이르는 급성 출혈을 일으키고 1시간 및 3시간 뒤에 신장 조직의 AQP2 mPNA 및 단백 발현을 각각 역전사 중합효소 연쇄반응법과 Western blot 분석법으로 조사하였다. 결과 : 출혈 후 1시간에 신장의 내수질에서 AQP2 mPNA 발혈은 유의하게 증가되었으나 AQP2 단백 이동 및 발현은 별화가 없?B다. 출혈 후 3시간에 AQP2 mRNA 발현 뿐 아니라 AQP2 단백 이동 및 발현이 모두 유의하게 증가하였다. 결론 : 혈액 종적 감소에 기인하여 신장에서 AQP2 수분 통로 발현이 증가되며 이는 체액용적 확보를 위한 보상 기전의 하나로서 작용하리라 추측되었다. Background : The present study was aimed to examine the regulation of aquaporin (AQP)-2 water channels in the kidney following blood volume depletion. Methods : Male Sprague-Dawley rats were acutely blood volume0depleted by withdrawal of arterial blood up to 2% of body weight. The expression of transcription - polymerase chain reaction and Westem blot analysis, respectively, in the inner medulla of the kidney 1 and 3 hours after the hemorrhage. Results : The mRNA expression if AQP2 was significantly increased 1 hour after thd bleeding However, neither the shuttling nor the total abundance of QQP2 proteins was significantly altered. On the contrary, 3 hours after the bleeding, the expression of AQP2 proteins as well as that of AQP2 mRNA was significantly incresaed. The shuttling of AQP2 proteins was also increased. Conclusion : These results sugest that an increased expression of AQP2 channels in the kidney may confer one of compensatory mechanisms restoring the circulation volume in an acute hypovolemic state.

흰쥐에서 급성 혈액 용적 감소에 따른 신조직의 Aquaporin-2 수분 통로 발현 변화

이영순 ( Ying Shun Li ),오윤화 ( Yoon Wha Oh ),이성수 ( Sung Su Lee ),이종은 ( Jong Un Lee ) 대한신장학회 2003 Kidney Research and Clinical Practice Vol.22 No.1

배경 : 양측 요관 패쇄 (bilateral ureteral ostruction; BUO)는 신장의 aquaporin-2 (AQP2)와 Na-K-2c1 cotransporter (NKCC2)의 발현을 감소기킨다고 알려져 있고 항이뇨호르몬의 투여는 신장의 AQP2 와 NKCC2의 발현을 증가시킨다고 알려져 있지만 BUO나 요로 패쇄 후 이뇨 (postobstructive diuresis; POD)상태에서 항이뇨호르몬을 투여했을 때의 효과에 대해서는 상세히 연구되어진 바가 없다. 본 연구에서는 24시간의 BUO를 만든 백서에서 POD-7 일째의 신장의 AQP2 와 NKCC2의 발현 및 항이뇨호르몬을 투여했을 때의 변화를 알아보고자 하였다. 방법 : Sprauge-Dawley 백서를 이용하여 Ⅰ군 (sham 수술 대조근)은 sham 수술을 시행한 대조군 (n=6), Ⅱ군 (BUO군)은 24시간의 BUO를 만든 후 해제한 군 (n=6), Ⅲ군 (dDAVP군)은 sham 수술호 dDAVP (1-deamino-8d-arginine vasopressin)를 osmotic minipump를 이용하여 시간당 20 ng의 속도로 지속적으로 7일간 피하 주사한 군 (n=6), Ⅳ군 (BUO+dDAVP군)은 24시간의 BUO를 만든 후 해제한 군 (n=6), Ⅲ군과 같은 방법으로 DDAVP를 투여한 군 (n=6)으로 하였다. 혈액 및 요 검사를 통해 네 군간의 생리학적 지표를 비교하였고 POD-7일에 해당하는 시점에서 네 군의 백서를 희생하여 면역조직화학적 검사와 Western blot 분석법을 이용하여 각 군의 신장의 AQP2와 NKCC2의 발현을 조사 비교하였다. 결과 : BUO군은 sham 수술 대조군에 비해 요량은 증가하였고 요 삼투질농도는 감소하였으며 (p<0.05) 신장의 AQP2 (29.1±4.2% vs. 100±10.0%, p<0.05)와 NKCC2 (40.2±5.4% vs. 100±7.9%, p<0.05)의 발현은 감소하였다. dDAVP군은 sham 수술 대조군에 비해 요량은 POD-5일까지 감소하였고 요 삼투질농도는 POD-7일까지 증가하였으며 (p<0.05) 신장의 AQP2 (206.5±19.0% vs. 100±10.0%, p<0.05)와 NKCC2 (180.6±13.3% vs. 100±7.9%, p<0.05)의 발현은 증가하였다. 그러나 BUO군은 BUO +dDAVP군과 비교하여 요량, 요삼투질농도에서 차이가 없었고 신장의 AQP2 (29.1±4.2% vs. 42.2±2.3%, p<0.84)와 NKCC2 (40.2±5.4% vs. 47.9±4.3%, p<0.91)의 발현에서도 차이가 없었다. 결론 : BUO와 POD는 신장의 AQP2와 NKCC2의 발현에 있어 감소를 보이고 장기간의 항이뇨호르몬의 투여에 대해 저항성을 나타내는데 이것이 다뇨와 요 삼투질농도의 감소와 같은 POD의 성상을 뒷받침하는 병태생리학적 기전이라 사료된다. Background : Bilateral ureteral obstuction (BUO) has been known to decrease the expression of renal aquaporin-2 (AQP2) and Na-K-2C1 cotransporter (NKCC2). The polyuria and urinary concentration defect in postobstructive diuresis (POD) may be explained by these molecular changes. By contrast chronic infusion of antidiuretic hormone (ADH) has been known to increase the expression of renal AQP2 and NKCC2, but there have been no studies regarding the chronic effect of ASH in molecular level when introducing to POD. We tried to identify the changes of renal expression of AQP2 and KNCC2 in 24 hour NUO rat at PID-7 day and the chronic effect of ADH to the expression of AQP2 and NKCC2 in sham operation rat and in 24 hour BUO rat, at sham operation 7 day and at POD-7 day, repectively. Methods : Twenty four Spraugue-Dawley rats were divided into four groups. Group Ⅰ (Control group) : sham operation rats(n=6). Group Ⅱ (BUO group) : 24 hour BUO and release of it (n=6). Group Ⅲ (dDAVP group) : dDAVP (1-dearmino-8d-arginine vasopressin : V2-receptot-selective agonist) infusion at rate of 20 ng/hour by somotic minipump subcutaneously for 7 days in sham operation rats (n=6) Group IV (BUO+dDAVP group) : dDAVO infusion at rate of 20 ng/hour by osmotic minipump by same method as Group Ⅲ in 24 hour BUO rats (n=6). All rats were sacrificed at POD-7 day (Group Ⅱ, Group Ⅳ) or sham operation-7 day (Group Ⅰ, Group Ⅲ) and renal expression of AQP2 and NKCC2 were analyzed by immunohistochemistry and by Western blot method. Blood and urinary chemistry examinat012ions were done concurrently. Results : BUO group showed increased urine output and decreased urine osmolality (p<0.05) and decreased expressions of AQP2 and NKCC2 compared with Control group {29.1±4.2% vs. 100±10.0% (p<0.05); 40.2±5.4% vs. 100±7.9% (p<0.05) respectively}. dDAVP group had decreased urine output from POD-1 day to POD-5 day and increased urine osmolaoity (p<0.05) POD-1 day to POD-7 day during and increased expressions of AQP2 and NKCC2 compared with Control group {206.5±19.0% vs. 100±10.0% (p<0.05); 180.6±13.3% vs. 100±7.9% (p<0.05) respectively}. But BUO group showed no difference in urine output and urine osmolality and expressions of AQP2 and NKCC2 compared with BUO+dDAVP group {29.1±4.2% vs. 42.2±2.3% (p<0.84); 40.2±5.4% vs. 47.9±4.3% (p<0.91) respectively}. Conclusion : BUO and POD show decreased expressions of AQP2 and NKCC2 and the unresponsiveness to chronic ADH infusion may be the pathophysiologic basis of POD such as increased urine output, decreased urine osmolality.