• The Korean Journal of Physiology and Pharmacology
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• 제5권3호
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• pp.231-241
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• 2001

The mammalian cortical collecting duct (CCD) plays a major role in regulating renal NaCl reabsorption, which is important in $Na^+$ and $Cl^-$ homeostasis. The M-1 cell line, derived from the mouse cortical collecting duct, has been used as a mammalian model of the study on the electrolytes transport in CCD. M-1 cells were grown on collagen-coated permeable support and short circuit current $(I_{sc})$ was measured. M-1 cells developed amiloride-sensitive current $5{\sim}7$ days after seeding. Apical and basolateral addition of ATP induced increase in $I_{sc}$ in M-1 cells, which was partly retained in $Na^+-free$ or $Cl^--free$ solution, indicating that ATP increased $Na^+$ absorption and $Cl^-$ secretion in M-1 cells. $Cl^-$ secretion was mediated by the activation of apical cystic fibrosis transmembrane regulator (CFTR) chloride channels and $Ca^{2+}-activated$ chloride channels, but $Na^+$ absorption was not mediated by activation of epithelal sodium channel (ENaC). ATP increased cAMP content in M-1 cells. The RT-PCR analysis demonstrated that M-1 cells express $P2Y_2,\;P2X_3\;and\;P2Y_4$ receptors. These results showed that ATP regulates $Na^+$ and $Cl^-$ transports via multiple P2 purinoceptors on the apical and basolateral membranes in M-1 cells.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1999년도 학술발표회 진행표 및 논문초록
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• pp.50-50
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• 1999

The mammalian cortical collecting duct (CCD) plays a major role in regulating renal NaCl absorption, which is important in controlling total body Na and Cl homeostasis. The M-1 cell line, derived from the mouse cortical collecting duct, is being used as a mammalian model of the CCD to study electrolytes transport.(omitted)

• 대한수의학회지
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• 제47권4호
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• pp.363-370
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• 2007

Mongolian gerbil (Meriones unguiculatus) has been as an model animal for studing the neurological disease such as stroke and epilepsy because of the congenital incompleteries in Willis circle, as well as the investigation of water metabolism because of the long time-survival in the condition of water-deprived desert condition, compared with other species animal. Aquaporin 2 (AQP2) expressed at the surface of principal cells in collecting duct results from an equilibrium between the AQP2 in intracellular vesicles and the AQP2 on the plasma membrane. Aquaporin 4 (AQP4), which is expressed in cell in a wide range of organ, is also present in the collecting duct principal cells where this is abundant in the basolateral plasma membranes and represent potential exit pathways from the cell for water entering via AQP2. In this research, we divide 3 groups of which each group include the 5 animals. In the study of 7 or 14 days water restricted condition, we investigated the AQP2 and AQP4 by using a quantitative immunohistochemistry in the kidney. The results obtained in this study were summarized as followings. AQP2 is abundant in the apical plasma membrane and apical vesicles in the collecting duct principal cell and at rare abundance in connecting tubules. In the water-deprived Mongolian gerbil kidney, expression of AQP2 was continuosly increased in the cortical collecting duct and inner medullary collecting duct. This increase was both the apical region and cytoplasm. AQP4 is mainly expressed in the inner medulla, although some expression is also noted in the more proximal segment. In the water-deprived Mongolian gerbil kidney, AQP4 was also increased in the inner medullary collecting duct. Immunoactivity was increased in entire inner medullary collecting duct and newly detected in cytoplasm of principal cell. These findings suggest that increased levels of AQP2 and AQP4 in the cortical and inner medulalry collecting duct may play a important role for maintain fluid balance in the water-deprived kidney.

• The Korean Journal of Physiology and Pharmacology
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• 제20권3호
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• pp.229-236
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• 2016

Resveratrol (RSV) may provide numerous protective effects against chronic inflammatory diseases. Due to local hypoxia and hypertonicity, the renal medulla is subject to extreme oxidative stress, and aldehyde products formed during lipid peroxidation, such as 4-hydroxy-2-hexenal (HHE), might be responsible for tubular injury. This study aimed at investigating the effects of RSV on renal and its signaling mechanisms. While HHE treatment resulted in decreased expression of Sirt1, AQP2, and nuclear factor erythroid 2-related factor 2 (Nrf2), mouse cortical collecting duct cells (M1) cells treated with HHE exhibited increased activation of p38 MAPK, extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and increased expression of NOX4, $p47^{phox}$, Kelch ECH associating protein 1 (Keap1) and COX2. HHE treatment also induced $NF-{\kappa}B$ activation by promoting $I{\kappa}B-{\alpha}$ degradation. Meanwhile, the observed increases in nuclear $NF-{\kappa}B$, NOX4, $p47^{phox}$, and COX2 expression were attenuated by treatment with Bay 117082, N-acetyl-l-cysteine (NAC), or RSV. Our findings indicate that RSV inhibits the expression of inflammatory proteins and the production of reactive oxygen species in M1 cells by inhibiting $NF-{\kappa}B$ activation.

• Applied Microscopy
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• 제37권3호
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• pp.167-174
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• 2007

Heat shock protein 70 (HSP70)은 다양한 질병상태와 치명적인 열 손상에서 세포 및 조직을 보호하는데 중요한 역할을 하며, 또한 외부의 stress로부터 세포내 단백질의 파괴와 변화를 감소시키는 단백질로 알려져 있다. 본 연구에서는 오랜 기간 동안 조혈기관의 치료제로 콩팥에서도 세포보호효과가 있는 것으로 알려진 Erythropoietin(EPO)을 투여하여 콩팥내의 HSP70의 발현변화를 세포수준에서 관찰하고자 하였다. Sprague-Dawley계 흰쥐를 사용하여 전자현미경적 면역조직화학법으로 rHuEPO투여군과 대조군에서 HSP70의 발현변화를 관찰하였다. 대조군에서 HSP70은 콩팥의 바깥수질과 속수질에서 관찰되었으며, 특히 속수질에서 강하게 발현되었고 그 부위는 속수질집합관세포와 헨레고리의 내림가는 부분이었다. EPO 투여군에서는 속수질과 바깥수질의 내림가는 부분에서는 발현변화가 관찰되지 않았으나, 바깥수질의 집합관 세포에서 발현이 급격히 증가함을 관찰되었다. 특히 대조군에서의 핵주변부위뿐 아니라 세포내 핵상부분을 비롯한 세포막주변부위에도 강한 면역 염색성을 나타내었다. 이러한 결과는 콩팥의 바깥수질에서 stress성 단백질인 HSP70의 조절기전이 EPO에 의해 매개됨을 보여주면, 세포 stress및 질병상태에서도 이러한 기전이 작용할 것으로 생각된다.

• Applied Microscopy
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• 제38권2호
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• pp.141-148
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• 2008

Nitric oxide는 콩팥에서 혈류의 조절, 사구체 혈액역동학 및 요세관의 운반에 중요한 역할을 하는 인자로서 최근 세포의 주기조절, 유사분열에도 관여하는 것으로 알려져 있다. 최근 연구 결과에 의하면 발생중인 콩팥에서의 nNOS의 발현은 성체와는 다른 것으로 알려져 있으나, 발생중인 콩팥에서의 발현 양상 및 발현시기 등에 대해서 알려진 바 없다. 따라서 본 연구에서는 발생중인 콩팥을 이용하여 분화중인 요세관에서 nNOS의 발현양상을 관찰하고자 하였다. 성체에서 nNOS의 면역반응성은 사구체 보우만주머니의 벽측상피와 치밀반, 내림가는부분 및 수질집합관에 발현되었다. 발생중인 콩팥에서는 임신 15일에 처음 발현하기 시작하였으며, 그 부위는 원위세관의 굵은오름부분이었다. 임신 20일에서 출산후 14일까지 새로이 형성되는 요세관의 굵은오름부분에서 관찰되었으나, 분화를 마친 요세관에서는 그 발현 양상이 감소하였다. 출생후 21일경에는 최종적으로 치밀반에만 국한되어 발현되었다. 속수질에서는 내림가는 부분과 속수질 집합관에서 발현되었으며 그 양상은 수질 중간부위에서 강하게 발현되었으며 시작부위와 말단부위에서는 약하게 발현되었다. 이상의 결과로 보아 굵은오름부분에서의 nNOS의 발현은 요세관의 분화정도와 일치하며 특히 그 발현양상이 분화가 활발한 시기에 강하게 발현되고 분화가 저하된 후에는 발현이 감소하는 것으로 보아 발생중인 콩팥의 성장에 중요한 역할을 할 것으로 생각된다.