Urea Restrains Aldosterone-Induced Development of Peanut Agglutinin–Binding on Embryonic Renal Collecting Duct Epithelia

Schumacher, Karl; Strehl, Raimund; Minuth, Will W.

doi:10.1097/01.asn.0000090744.88722.ff

Cited by 6 publications

(6 citation statements)

References 27 publications

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“…Lack of immunolabel for mab Ki67 reveals in analogy to the adult kidney that the cells within the epithelium stop to divide after day 6 of perfusion culture and remain in the functional interphase (data not shown). As demonstrated in recent investigations the generation of a flat CD epithelium with 6 mm in diameter derived from renal CD ampulla cells appears as a reliable model to investigate basic cellbiological questions such as epithelial polarization, upregulation, modulation and maintenance of CD features [18,22].…”

Section: Resultsmentioning

confidence: 99%

“…1D). The holders were then placed for 24 hours in a 24-well plate in a CO 2 -incubator containing 5% CO 2 /95% air atmosphere [17,18]. During culture of the explants in Iscove´s Modified Dulbecco´s Medium (IMDM; order # 21980 -032; Gibco BRL Life Technologies, Eggenstein, Germany) including 10% fetal bovine serum (Boehringer, Mannheim, Germany) an outgrowth of cells from the CD ampullae was observed.…”

Section: Generation Of Flat CD Epithelia In Perfusion Culturementioning

confidence: 99%

“…Aldosterone was applied at a concentration of 1 x 10 -7 M (SigmaAldrich-Chemie, Deisenhofen, Germany) and 1% antibioticantimycotic solution (Gibco BRL-Life Technologies) was added to all culture media. Furthermore, up to 50 mmol/l HEPES (Gibco BRL-Life Technologies) was used in the medium to maintain a constant pH of 7.4 in perfusion culture under atmospheric air containing 0.3% CO 2 [18].…”

Section: Generation Of Flat CD Epithelia In Perfusion Culturementioning

confidence: 99%

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Generation of Renal Tubules at the Interface of an Artificial Interstitium

Minuth

Sorokin

Schumacher

2004

Cell Physiol Biochem

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During kidney development a multitude of tubular portions is formed. Little knowledge is available by which cellbiological mechanism a cluster of embryonic cells is able to generate the threedimensional structure of a tubule. However, this know-how is most important in tissue engineering approaches such as the generation of an artificial kidney module or for the therapy of renal diseases using stem cells. To obtain cellbiological insights in parenchyme development we elaborate a new technique to generate under in vitro conditions renal tubules derived from the embryonic cortex of neonatal rabbits. The aim of the experiments is to establish a specific extracellular environment allowing optimal threedimensional development of renal tubules under serum-free culture conditions. In the present paper we demonstrate features of the renal stem cell niche and show their isolation as intact microcompartiments for advanced tissue culture. Perfusion culture in containers exhibiting a big dead fluid volume results in the development of a flat collecting duct (CD) epithelium at the surface of the tissue explant. In contrast, by fine-tuning the dead fluid volume within a perfusion culture container by an artificial interstitium made of a polyester fleece shows the generation of tubules. It is an up to date unknown morphogenetic information which tells the cells to form tubular structures.

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Section: Resultsmentioning

confidence: 99%

Section: Generation Of Flat CD Epithelia In Perfusion Culturementioning

confidence: 99%

Section: Generation Of Flat CD Epithelia In Perfusion Culturementioning

confidence: 99%

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Generation of Renal Tubules at the Interface of an Artificial Interstitium

Minuth

Sorokin

Schumacher

2004

Cell Physiol Biochem

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“…In addition, we found that embryonic renal tissue is a target for aldosterone, since it promotes the functional maturation of collecting duct epithelium [27]. Furthermore, we detected that aldosterone exerts a tubulogenic effect promoting the development of structured tubules derived from renal stem/progenitor cells [18][19][20].…”

Section: Discussionmentioning

confidence: 79%

The tubulogenic effect of aldosterone is attributed to intact binding and intracellular response of the mineralocorticoid receptor

Minuth

Denk

et al. 2007

Open Life Sciences

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Abstract:Little is known about the extra-and intracellular stimuli inducing renal stem/progenitor cells to develop into three-dimensionally structured tubules. To study this specific development in a controlled environment, we used an advanced culture technique. Embryonic tissue derived from neonatal rabbit kidney was placed in a perfusion culture container at the interface of an artificial interstitium made of a polyester fleece. Culture was carried out in chemically defined Iscove's Modified Dulbecco's Medium (IMDM) for 13 days. Development of tubules was histochemically detected on cryosections labeled with Soybean Agglutinin (SBA). The experiments showed that aldosterone exerts a specific tubulogenic effect. Application of aldosterone (1 × 10 −7 M) raised numerous SBA-labeled tubules, while in the absence of the steroid hormone the development of tubules was lacking. Specificity of hormone action was analyzed by the use of aldosterone antagonists. Administration of spironolactone (1 × 10 −4 M) and canrenoate(1 × 10 −5 M) completely inhibited the development of tubules. Finally, disrupting the intracellular molecular complex of the mineralocorticoid receptor (MR) and heat shock proteins by geldanamycin (2 µg/ml) prevented the development of tubules. Our results suggest that the tubulogenic effect induced by aldosterone is attributed to both hormone binding and an undisturbed intracellular response of the MR.

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“…23 In perfusion culture the cells of the CD ampulla have been shown to develop into a functional epithelium showing features of adult principal and intercalated cells. 17 It has been shown earlier that the development of embryonic collecting duct cells can be influenced by the application of aldosterone [24][25][26] as well as by urea 27 and increased electrolyte concentrations. 28 Astonishingly, the present experiments show that chronically decreasing oxygen partial pressure in a defined manner during culture can trigger similar changes in the differentiation profile of the embryonic epithelia.…”

Section: Discussionmentioning

confidence: 99%

Controlled Respiratory Gas Delivery to Embryonic Renal Epithelial Explants in Perfusion Culture

2004

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During generation of artificial tissues high levels of oxygen are usually available whereas after implantation into a recipient's body the implant is not vascularized immediately, which leads to low oxygen partial pressures within the implanted tissue. Under these conditions cells will experience an oxygen shortage, contrasting with the abundance of oxygen during culture. It is uncertain whether tissues can be trained to tolerate such an acute hypoxic situation so that nonphysiological stress reactions and tissue necrosis can be avoided. To investigate the effects of varying oxygen levels on embryonic renal tissue in vitro we have been developing a model system combining continuous medium renewal with the ability to control levels of oxygen and carbon dioxide by gas equilibration through gas-permeable tubing. Renal embryonic tissue from neonatal rabbit was cultured in serum-free Iscove's modified Dulbecco's medium at 45, 90, 115, and 160 mmHg oxygen partial pressure for 14 days under continuous medium exchange in such a setup. After a 14-day culture period tissue sections were analyzed by cell biological methods and compared with fresh tissue histology. Surprisingly, embryonic renal explants survive and maintain good morphology for 14 days under all O(2) conditions tested. Expression of cytokeratin 19 within the established epithelium remains unchanged, indicating a structurally intact tissue. However, Na/K-ATPase is clearly downregulated under low O(2) conditions, whereas COX-2 expression increases drastically. An antiparallel effect of decreased O(2) concentrations on glycoprotein expression can be demonstrated with the lectin Dolichos biflorus agglutinin. Scanning electron microscopy reveals oxygen-dependent changes in cellular surface differentiation of developed collecting duct epithelium.

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Urea Restrains Aldosterone-Induced Development of Peanut Agglutinin–Binding on Embryonic Renal Collecting Duct Epithelia

Cited by 6 publications

References 27 publications

Generation of Renal Tubules at the Interface of an Artificial Interstitium

Generation of Renal Tubules at the Interface of an Artificial Interstitium

The tubulogenic effect of aldosterone is attributed to intact binding and intracellular response of the mineralocorticoid receptor

Controlled Respiratory Gas Delivery to Embryonic Renal Epithelial Explants in Perfusion Culture

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