Expression of mRNA for natriuretic peptide receptor subtypes in bovine kidney

1994 ◽  
Vol 267 (2) ◽  
pp. F318-F324 ◽  
Author(s):  
T. Yamamoto ◽  
L. Feng ◽  
T. Mizuno ◽  
S. Hirose ◽  
K. Kawasaki ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Natriuretic Peptide ◽  
Collecting Duct ◽  
Biologically Active ◽  
Ribonuclease Protection Assay ◽  
Receptor Subtypes ◽  
Total Rna ◽  
Duct Cells ◽  
Glomerular Epithelial Cells ◽  
Collecting Duct Cells

The localization of mRNA for atrial natriuretic peptide (ANP) receptor subtypes (A, B, C) in the kidney was examined. Quantitative analysis of the ribonuclease protection assay showed that the numbers of type A receptor (ANPRA) mRNA were 6.9 x 10(7) in the glomeruli and 10.4 x 10(7) molecules/micrograms of total RNA in the inner medulla, and that of type C receptor (ANPRC) mRNA was 21.7 x 10(7) molecules/micrograms of total RNA in the glomeruli. The type B receptor (ANPRB) mRNA was present in smaller numbers (4.5-4.9 x 10(6) molecules/micrograms of total RNA) evenly throughout the kidney fractions. In situ hybridization demonstrated both ANPRA and ANPRC mRNA selectively in the glomerular epithelial cells and ANPRA mRNA in the collecting duct cells of the inner medulla. ANPRC was also localized on the foot processes of glomerular epithelial cells by immunohistochemistry using a specific antibody against the receptor. These results indicate that ANPRA is the major biologically active receptor for the ANP family of hormones in the kidney and is present selectively on the glomerular epithelial cells and inner medullary collecting duct cells. These cells are presumed to play a role in the regulation of glomerular filtration rate and sodium excretion induced by the family of ANP.

Expression of AQP family in rat kidneys during development and maturation

1997 ◽  
Vol 272 (2) ◽  
pp. F198-F204 ◽  
Author(s):  
T. Yamamoto ◽  
S. Sasaki ◽  
K. Fushimi ◽  
K. Ishibashi ◽  
E. Yaoita ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Kidney Development ◽  
Collecting Duct ◽  
Rat Kidney ◽  
Staining Intensity ◽  
Ribonuclease Protection Assay ◽  
Duct Cells ◽  
Apical Side ◽  
Basolateral Side ◽  
Collecting Duct Cells

The mRNA expression and localization of the aquaporin (AQP) family in rat kidney were examined by ribonuclease protection assay and immunohistochemistry. AQP1, AQP2, AQP3, and AQP4 mRNA were hardly detectable in 16-day gestation fetuses. AQP1 mRNA was explosively expressed at 1 wk, keeping the level throughout life. AQP2 mRNA expression was apparently noticed in 18-day fetuses and was enhanced gradually with age to reach a plateau at 4 wk. AQP3 and AQP4 mRNA expression was significantly found at birth but was not changed remarkably thereafter. AQP2 protein appeared first at the apical side of collecting duct cells in 18-day fetuses. The staining intensity at the site increased with age, and basolateral staining was added in adult rats. AQP3 was distinctly demonstrated at the basolateral side of collecting duct cells after birth, and the staining intensity was almost stable throughout life. The progressive induction of AQP2 expression in the first 4 wk after birth is presumed to contribute to the maturation of urinary concentrating capacity during the kidney development.

scholarly journals Telomerase immortalization of principal cells from mouse collecting duct

2010 ◽  
Vol 299 (6) ◽  
pp. F1507-F1514 ◽  
Author(s):  
Stacy L. Steele ◽  
Yongren Wu ◽  
Robert J. Kolb ◽  
Monika Gooz ◽  
Courtney J. Haycraft ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Collecting Duct ◽  
Whole Cell ◽  
Renal Epithelial Cells ◽  
Principal Cells ◽  
Duct Cells ◽  
Collecting Duct Cells ◽  
Well Differentiated

Recently, the use of overexpression of telomerase reverse transcriptase (TERT) has led to the generation of immortalized human cell lines. However, this cell immortalization approach has not been reported in well-differentiated mouse cells, such as renal epithelial cells. We sought to establish and then characterize a mouse collecting duct cell line, using ectopic expression of mTERT. Isolated primary cortical collecting duct (CCD) cell lines were transduced with mouse (m)TERT, using a lentiviral vector. mTERT-negative cells did not survive blasticidin selection, whereas mTERT-immortalized cells proliferated in selection media for over 40 subpassages. mTERT messenger RNA and telomerase activity was elevated in these cells, compared with an SV40-immortalized cell line. Flow cytometry with Dolichos biflorus agglutinin was used to select the CCD principal cells, and we designated this cell line mTERT-CCD. Cells were well differentiated and exhibited morphological characteristics typically found in renal epithelial cells, such as tight junction formation, microvilli, and primary cilia. Further characterization using standard immunofluorescence revealed abundant expression of aquaporin-2 and the vasopressin type 2 receptor. mTERT-CCD cells exhibited cAMP-stimulated/benzamil-inhibited whole cell currents. Whole cell patch-clamp currents were also enhanced after a 6-day treatment with aldosterone. In conclusion, we have successfully used mTERT to immortalize mouse collecting duct cells that retain the basic in vivo phenotypic characteristics of collecting duct cells. This technique should be valuable in generating cell lines from genetically engineered mouse models.

scholarly journals Pax2 and Pax8 Proteins Regulate Urea Transporters and Aquaporins to Control Urine Concentration in the Adult Kidney

2020 ◽  
Vol 31 (6) ◽  
pp. 1212-1225 ◽  
Author(s):  
Ann M. Laszczyk ◽  
Atsuko Y. Higashi ◽  
Sanjeevkumar R. Patel ◽  
Craig N. Johnson ◽  
Abdul Soofi ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Target Genes ◽  
Collecting Duct ◽  
Expression Patterns ◽  
Renal Medulla ◽  
Urine Concentration ◽  
Renal Epithelia ◽  
Duct Cells ◽  
Collecting Duct Cells ◽  
Collecting Tubules

BackgroundAs the glomerular filtrate passes through the nephron and into the renal medulla, electrolytes, water, and urea are reabsorbed through the concerted actions of solute carrier channels and aquaporins at various positions along the nephron and in the outer and inner medulla. Proliferating stem cells expressing the nuclear transcription factor Pax2 give rise to renal epithelial cells. Pax2 expression ends once the epithelial cells differentiate into mature proximal and distal tubules, whereas expression of the related Pax8 protein continues. The collecting tubules and renal medulla are derived from Pax2-positive ureteric bud epithelia that continue to express Pax2 and Pax8 in adult kidneys. Despite the crucial role of Pax2 in renal development, functions for Pax2 or Pax8 in adult renal epithelia have not been established.MethodsTo examine the roles of Pax2 and Pax8 in the adult mouse kidney, we deleted either Pax2, Pax8, or both genes in adult mice and examined the resulting phenotypes and changes in gene expression patterns. We also explored the mechanism of Pax8-mediated activation of potential target genes in inner medullary collecting duct cells.ResultsMice with induced deletions of both Pax2 and Pax8 exhibit severe polyuria that can be attributed to significant changes in the expression of solute carriers, such as the urea transporters encoded by Slc14a2, as well as aquaporins within the inner and outer medulla. Furthermore, Pax8 expression is induced by high-salt levels in collecting duct cells and activates the Slc14a2 gene by recruiting a histone methyltransferase complex to the promoter.ConclusionsThese data reveal novel functions for Pax proteins in adult renal epithelia that are essential for retaining water and concentrating urine.

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