scholarly journals Evaluation of the Role of Na+/ K+ ATPase Ion Transporters and Na+/K+/ 2Cl and Na+/H+ Exchanger Cotransporters in Nephrons of Periophthalmus Waltoni Using Immunohistochemistry and Histology

2021 ◽  
Author(s):  
Kave Esfandiari ◽  
Mohammad Babaei ◽  
Mina Amiri-Farahani ◽  
Ali Kalantari-Hesari ◽  
Hassan Morovvati
Keyword(s):  
Epithelial Cells ◽  
Kidney Tissue ◽  
Proximal Tubules ◽  
Renal Tubules ◽  
Ion Regulation ◽  
Renal Histology ◽  
Collecting Ducts ◽  
Distal Tubules ◽  
Periophthalmus Waltoni

Abstract Kidneys play an important role in regulating the balance of water and ions in freshwater and seawater fish. However, complex kidney structures impair a comprehensive understanding of kidney function. In this study, in addition to renal histology, Na+/K+/ATPase ion transporter proteins and Na+/K+/2Cl− and NHE3 cotransporters were located in Priophthalmus waltoni kidney tissue to evaluate the ion regulation abilities of epithelial cells in various parts of nephrons. The renal tubules are composed of proximal tubules and distal tubules, followed by collecting tubes and finally collecting ducts. Light microscope immunohistochemistry was utilized to locate Na+/ K+-ATPase along renal tubules and collecting ducts. However, the distribution of the Na+/K+-ATPase immune response varies in different sections. Na+/K+/CL− cotransporter positioning was reported only in collecting tubes and collecting ducts, and proximal tubes and distal tubes did not respond to Na+/K+/Cl− cotransporter immunolocalization. Immunohistochemical response for NHE3 localization was detected only at the apex of epithelial cells of proximal tubules and collecting tubes. The distal tubes showed negative reaction and the collecting ducts showed a weak response to NHE3 safety immunolocalization.

Ochratoxin A and Citrinin Induced Nephrosis in Beagle Dogs II. Pathology

1977 ◽  
Vol 14 (3) ◽  
pp. 261-272 ◽  
Author(s):  
D. N. Kitchen ◽  
W. W. Carlton ◽  
J. Tuite
Keyword(s):  
Ochratoxin A ◽  
Proximal Tubules ◽  
Lymphoid Tissues ◽  
Tubular Epithelial Cells ◽  
Beagle Dogs ◽  
Peripheral Lymph ◽  
Collecting Ducts ◽  
Gross Lesions ◽  
Colon And Rectum ◽  
Distal Tubules

Beagle dogs were given ochratoxin A (0.1 and 0.2 mg/kg) and citrinin (5 and 10 mg/kg) alone and in two dose combinations for 14 days. The gross lesions included focal peritonitis and intestinal intussusceptions in dogs given citrinin. Changes in the kidneys of dogs given ochratoxin A were degeneration and necrosis with desquamation of tubular epithelial cells, primarily in the straight segment of the proximal tubules. Dogs given 10 mg/kg citrinin had similar changes in the distal tubules and collecting ducts. Dogs given combined doses of citrinin and ochratoxin A had degeneration and necrosis in proximal and distal tubules, and in thin segments and the collecting ducts; there were desquamated cells and granular casts in the lumina. Dogs given ochratoxin A had necrosis of lymphoid tissues in the spleen, tonsil, thymus, peripheral lymph nodes and lymph nodules of the ileum, colon and rectum. There was ulceration of the mucosa of the intestine in dogs given large combined doses of ochratoxin A and citrinin.

Return of the secretory kidney

2002 ◽  
Vol 282 (1) ◽  
pp. F1-F9 ◽  
Author(s):  
Jared J. Grantham ◽  
Darren P. Wallace
Keyword(s):  
Glomerular Filtration ◽  
Extracellular Fluid ◽  
Fluid Secretion ◽  
Proximal Tubules ◽  
Renal Tubules ◽  
Functional Elements ◽  
Dry Land ◽  
Fluid Reabsorption ◽  
Collecting Ducts ◽  
Urine Formation

The evolution of the kidney has had a major role in the emigration of vertebrates from the sea onto dry land. The mammalian kidney has conserved to a remarkable extent many of the molecular and functional elements of primordial apocrine kidneys that regulate fluid balance and eliminate potentially toxic endogenous and xenobiotic molecules in the urine entirely by transepithelial secretion. However, these occult secretory processes in the proximal tubules and collecting ducts of mammalian kidneys have remained underappreciated in the last half of the twentieth century as investigators focused, to a large extent, on the mechanisms of glomerular filtration and tubule sodium chloride and fluid reabsorption. On the basis of evidence reviewed in this paper, we propose that transepithelial salt and fluid secretion mechanisms enable mammalian renal tubules to finely regulate extracellular fluid volume and composition day to day and maintain urine formation during the cessation of glomerular filtration.

FC 014INFLUENCE OF GENETIC VARIATION IN SLC7A13/AGT1 IN HUMAN CYSTINURIA

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Ria Schönauer ◽  
Anna Seidel ◽  
Linda Pöschla ◽  
Elena Hantmann ◽  
Soumeya Bekri ◽  
...  
Keyword(s):  
Complex Formation ◽  
Proximal Tubules ◽  
Patient Specific ◽  
Incomplete Penetrance ◽  
Renal Tubules ◽  
Pathogenic Variants ◽  
Cystine Transporter ◽  
S3 Segment

Abstract Background and Aims Cystinuria (CU) is an inherited renal disorder based on urinary wasting of dibasic amino acids, urinary precipitation, and consecutive cystine stone formation. It is caused by pathogenic variants in two distinct disease genes, SLC3A1 and SLC7A9, both of which encode subunits of a heterodimeric tubular amino acid transporter, rBAT/SLC3A1 and BAT1/SLC7A9, located at the apical membrane of proximal renal tubules. CU is marked by incomplete penetrance and substantial disease variability. Recently, a novel cystine transporter, consisting of the light chain AGT1/SLC7A13 and its heterodimeric partner rBAT/SLC3A1 has been identified in the S3 segment of murine proximal tubules. In this study, we aim at evaluating the role of AGT1 in cystinuric patients with or without mutations in either SLC3A1 or SLC7A9, analyzing the role of AGT1/SLC7A13 as novel disease gene or genetic modifier in CU. Method A multicenter European CU-cohort comprising 132 individuals was screened for pathogenic variants in SLC3A1, SLC7A9, and SLC7A13 using high-throughput multiplex PCR-based amplification and next-generation sequencing (MiSeq Illumina) followed by multiplex ligation-dependent probe amplification (MLPA) of SLC3A1 and SLC7A9. For functional in vitro studies, epitope-tagged human and murine rBAT and AGT1 proteins were transiently expressed in different cell systems. Heterodimer complex formation was analyzed by co-immunoprecipitation and western blot studies and membrane trafficking was evaluated by immunofluorescence microscopy. Results Genectic analysis of our CU-cohort did not reveal indiviuals with SLC7A13 variation only, however we found three patients harbouring heterozygous missense variants in addition to pathogenic or VUS variants in SLC3A1 or SLC7A9. To evaluate their influence on the generation of functional cystine transporters in vitro, different cell models were transiently transfected with plasmids expressing wildtype or mutant proteins. In line with previous reports, co-expression of AGT1 and rBAT wildtype allowed efficient complex formation as AGT1-induced maturation of rBAT was detected by increased mature N-glycosylation, co-immunoprecipitation and membrane insertion. Whereas AGT1 patient variants p.Met452Thr (SLC7A13 c.1355T>C) and p.Ile174Phe (SLC7A13 c.520A>T) behaved comparable to wildtype AGT1, variants p.Asn45Lys (SLC7A13 c.135C>G) and p.Leu270Phe (SLC7A13 c.808C>T) led to clearly reduced glycosylation patterns and trafficking deficits of rBAT wildtype protein. Next, the mutual influence of pathogenic variation in both, AGT1 and rBAT, will unravel the consequences of patient-specific molecular interactions on the functional expression of cystine transporter complexes. Conclusion Here, we report three CU-patients with variants in SLC7A13 combined with either SLC3A1 or SLC7A9. For two of these variants, in vitro functional analysis revealed pathogenic molecular mechanisms disturbing complex formation, maturation and trafficking of rBAT. We hypothesize that specific pathogenic variants in SLC7A13 interfere with efficient membrane localization of heterodimeric cystine transporters, which results in modulation of cystine transport in the S3 segment of proximal tubules in CU-patients.

Renal excretion of uric acid in the rat: a micropuncture and microperfusion study

1976 ◽  
Vol 230 (3) ◽  
pp. 768-776 ◽  
Author(s):  
F Roch-Ramel ◽  
F Diezi-Chomety ◽  
D De Rougemont ◽  
M Tellier ◽  
J Widmer ◽  
...  
Keyword(s):  
Uric Acid ◽  
Renal Excretion ◽  
Proximal Tubules ◽  
Free Flow ◽  
Collecting Ducts ◽  
Order Of Magnitude ◽  
Plasma Urate ◽  
Distal Tubules

Free-flow micropuncture experiments were done in rats of three strains infused with small amounts of urate [plasma urate (P urate) = 95 +/- 8 muM]. Urate concentrations in tubular fluid were measured by an accurate chemical fluorometric ultramicromethod. In fluid from surface glomeruli, the glomerular fluid-to-plasma urate ratio [GF/P) urate] was 0.99 +/- 0.03 (n=11), i.e., lower than expected for total ultrafiltrability of plasma urate. Along proximal convolutions, net reabsorption of 55% of filtered urate was demonstrated. Small amounts of urate may have been reabsorbed between late proximal and early distal sites. Net transepithelial movements of urate did not occur in distal tubules or collecting ducts. In microperfusion experiments on proximal tubules, both a reabsorptive flow of urate (loss of perfused [2-14C]urate) and a secretory flow (entrance of cold urate into perfusate) of the same order of magnitude were demonstrated. Neither flow was influenced by simultaneous water movements. Microperfusion of Henle's loops indicated a significant but very small net reabsorption.

Characterization and localization of side population (SP) cells in zebrafish kidney hematopoietic tissue

Blood ◽  
2008 ◽  
Vol 111 (3) ◽  
pp. 1131-1137 ◽  
Author(s):  
Isao Kobayashi ◽  
Kazuyuki Saito ◽  
Tadaaki Moritomo ◽  
Kyosuke Araki ◽  
Fumio Takizawa ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Renal Tubule ◽  
Side Population ◽  
Flow Cytometric Analysis ◽  
Kidney Tissue ◽  
Renal Tubules ◽  
Hematopoietic Tissue ◽  
Hematopoietic Stem ◽  
Hoechst Dye ◽  
And Function

Abstract We previously showed that side population (SP) cells, characterized by specific Hoechst dye efflux pattern in flow cytometric analysis, were present in teleost kidney hematopoietic tissue, and that kidney SP cells were enriched in hematopoietic stem cells (HSCs). ABCG2/Abcg2 is an ATP-binding cassette (ABC) transporter that is known to be associated with Hoechst dye efflux activity of mammalian HSCs. In the present study, we examined the expression and function of Abcg2 in kidney SP cells from zebrafish (Danio rerio). Although the zebrafish genome has 4 paralogous copies of ABCG2 (zAbcg2a, b, c, and d), zAbcg2a and zAbcg2c mRNA was expressed in kidney SP cells. Transfection of COS-7 cells with zAbcg2a and zAbcg2c showed that zAbcg2a was directly associated with the SP phenotype. These results indicate that zAbcg2a mRNA is a useful marker for zebrafish HSCs. In situ hybridization in kidney tissue showed that zAbcg2a-positive cells were sporadically localized on the surface of renal tubules, and tightly adhered to renal tubule epithelial cells. This result suggests that teleost HSCs adhere to the surface of renal tubules, and that renal tubule epithelial cells are a key component of HSC niche in teleosts.

scholarly journals Light-Microscopic Immunocytochemistry for Gentamicin and Its Use for Studying Uptake of the Drug in Kidney

2009 ◽  
Vol 53 (8) ◽  
pp. 3302-3307 ◽  
Author(s):  
Kunio Fujiwara ◽  
Masashi Shin ◽  
Hayato Matsunaga ◽  
Tetsuya Saita ◽  
Lars-Inge Larsson
Keyword(s):  
Renal Toxicity ◽  
Rat Kidney ◽  
Proximal Tubules ◽  
Proximal Tubule Cells ◽  
Collecting Ducts ◽  
Immunocytochemical Method ◽  
Tubule Cells ◽  
Distal Tubules ◽  
Pretreatment Conditions ◽  
Single Intravenous Administration

ABSTRACT Gentamicin (GM) is a widely used antibiotic but shows renal toxicity. We produced a serum against GM (anti-GM) conjugated to bovine serum albumin with N-(gamma-maleimidobutyryloxy)succinimide. The antiserum was monospecific for GM and did not cross-react with the analog streptomycin, tobramycin, kanamycin, or amikacin. The antiserum also detected glutaraldehyde-fixed GM, and this enabled us to develop an immunocytochemical method for detecting the uptake of GM in rat kidney. Twelve hours after a single intravenous administration of GM, immunocytochemistry revealed that GM accumulated in the S1, S2, and S3 segments of the proximal tubules, as well as in the distal tubules and collecting ducts. By 12 h after injection, the drug was detected in cytoplasmic granules of the proximal tubule cells. However, early (1 h) after injection, drug accumulation was detected in the microvilli of these cells. The distal tubules and collecting ducts contained scattered swollen cells, reminiscent of necrotic cells, in which both the nuclei and the cytoplasm reacted strongly with GM. No staining occurred in the kidneys of saline-injected control rats. These results agree with previous studies showing that GM is endocytosed in the proximal tubules and accumulates in lysosomes. Additionally, our results show that GM also accumulates in the distal tubules and collecting ducts. This was achieved by systematically varying the pretreatment conditions—an approach necessary for detecting GM in different subcellular compartments. This approach should be useful for accurately detecting the uptake and toxicity of the antibiotic in different tissues.

scholarly journals Structural and ultrastructural study of the rabbit kidney exposed to carbamate insecticide

2014 ◽  
Vol 83 (4) ◽  
pp. 295-298 ◽  
Author(s):  
Viera Almášiová ◽  
Katarína Holovská ◽  
Viera Cigánková
Keyword(s):  
Epithelial Cells ◽  
Normal Structure ◽  
Degenerative Changes ◽  
Rabbit Kidney ◽  
Kidney Cortex ◽  
Histological Structure ◽  
Collecting Ducts ◽  
Distal Tubules ◽  
Tubular Sections ◽  
Collecting Tubules

The aim of the present study was to determine the influence of orally administered insecticide bendiocarb on the structure and ultrastructure of the kidney parenchyma in rabbits. Bendiocarb in the form of capsules (96% Bendiocarb, Bayer), at a dose of 5 mg/kg of body weight was fed daily for 3 days. After sampling, kidney sections of experimental and control animals were evaluated. Under a light and electron microscope the diffuse degenerative changes in kidney cortex and medulla were noted. Light microscopy revealed that the renal corpuscles had normal structure, but other nephron components and the collecting ducts were invariably changed. The epithelial cells inside the proximal and distal tubules and collecting ducts possessed increased quantity of cytoplasmic vacuoles and some tubular sections showed cellular sloughing and necrotization. The cells within the thin limbs of the Henley’s loops had normal histological structure except for sporadic necrotizing cells within some segments. The ultrastructural evaluation showed extensive cytoplasmic vacuolisation and degenerative changes, such as mitochondrial swelling and shortening of basal infoldings within proximal and distal tubules, and microvilli reduction within proximal tubules. Cells of the collecting tubules exhibited a higher number of vacuoles and some cells had apparently reduced organelles. The cells of the thin limbs of the Henle’s loop showed more vacuolised cytoplasm, some tubular sections revealed cellular detachment between the adjacent epithelial cells and rare necrotising epithelial cells were observed. The described findings addressed in the present study indicate an adverse effect of bendiocarb on the kidney parenchyma in rabbits.

scholarly journals Outbreaks of Renal Failure Associated with Melamine and Cyanuric Acid in Dogs and Cats in 2004 and 2007

2007 ◽  
Vol 19 (5) ◽  
pp. 525-531 ◽  
Author(s):  
Cathy A. Brown ◽  
Kyu-Shik Jeong ◽  
Robert H. Poppenga ◽  
Birgit Puschner ◽  
Doris M. Miller ◽  
...  
Keyword(s):  
Renal Failure ◽  
Cyanuric Acid ◽  
Interstitial Fibrosis ◽  
Renal Tissue ◽  
Proximal Tubules ◽  
Hepatic Enzyme ◽  
Pet Food ◽  
Histologic Change ◽  
Collecting Ducts ◽  
Distal Tubules

Sixteen animals affected in 2 outbreaks of pet food-associated renal failure (2 dogs in 2004; 10 cats and 4 dogs in 2007) were evaluated for histopathologic, toxicologic, and clinicopathologic changes. All 16 animals had clinical and laboratory evidence of uremia, including anorexia, vomiting, lethargy, polyuria, azotemia, and hyperphosphatemia. Where measured, serum hepatic enzyme concentrations were normal in animals from both outbreaks. All animals died or were euthanized because of severe uremia. Distal tubular lesions were present in all 16 animals, and unique polarizable crystals with striations were present in distal tubules or collecting ducts in all animals. The proximal tubules were largely unaffected. Crystals and histologic appearance were identical in both outbreaks. A chronic pattern of histologic change, characterized by interstitial fibrosis and inflammation, was observed in some affected animals. Melamine and cyanuric acid were present in renal tissue from both outbreaks. These results indicate that the pet food-associated renal failure outbreaks in 2004 and 2007 share identical clinical, histologic, and toxicologic findings, providing compelling evidence that they share the same causation.

Immunolocalization of aquaporin-8 in rat kidney, gastrointestinal tract, testis, and airways

2001 ◽  
Vol 281 (6) ◽  
pp. F1047-F1057 ◽  
Author(s):  
Marie-Louise Elkjær ◽  
Lene N. Nejsum ◽  
Veronika Gresz ◽  
Tae-Hwan Kwon ◽  
Uffe B. Jensen ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Southern Blotting ◽  
Rat Kidney ◽  
Cell Types ◽  
Myoepithelial Cells ◽  
Proximal Tubules ◽  
Basal Cells ◽  
Rt Pcr ◽  
Collecting Ducts ◽  
Outer Stripe

First published August 8, 2001; 10.1152/ajprenal.00158.2001.—The purpose of this study was to determine the cellular and subcellular localization of aquaporin-8 (AQP8) in rat kidney and other organs by RT-PCR analyses and by immunoblotting and immunohistochemistry using peptide-derived rabbit antibodies to rat AQP8. RT-PCR and Southern blotting revealed the presence of AQP8 mRNA in all kidney zones. LLC-PK1 cells transfected with a rat AQP8 construct exhibited strong labeling with the affinity-purified antibodies, whereas controls using cells transfected with the vector, but without the insert, were negative. The labeling was almost exclusively associated with intracellular vesicles. Immunoblotting of kidney membrane fractions revealed a predominant single band of 26–28 kDa. AQP8 immunoreactivity was mainly present in the cortex and outer stripe of the outer medulla. Sequential ultracentrifugation of rat kidney membrane revealed that AQP8 resides predominantly in intracellular vesicular fractions. Immunocytochemistry revealed modest labeling of proximal tubules and weak labeling of collecting ducts in cortex and medulla of rat kidney. The labeling was confined to cytoplasmic areas with no labeling of the brush border. Immunoblotting and RT-PCR/Southern blotting also revealed the presence of AQP8 protein and mRNA in rat liver, testis, epididymis, duodenum, jejunum, colon, and bronchi/trachea. Consistent with this, immunohistochemistry revealed AQP8 labeling in the hepatocytes and spematogenic cells in testis and in the basal cells in ductus epididymis, trachea, and bronchial epithelia. Moreover, AQP8 labeling was observed in the myoepithelial cells in salivary, bronchial, and tracheal glands with no labeling of acini or ductal epithelial cells. AQP8 is also present in the surface epithelial cells in duodenum, jejunum, and colon. In conclusion, AQP8 is expressed at low levels in rat kidney proximal tubules and collecting ducts, and it is present in distinct cell types in liver, testis, epididymis, duodenum, jejunum, colon, trachea, and principal bronchi as well as in multiple glands, including salivary glands.

The effect of chronic hydrochlorothiazide administration on renal function in the rat

1986 ◽  
Vol 70 (4) ◽  
pp. 379-387 ◽  
Author(s):  
S. J. Walter ◽  
D. G. Shirley
Keyword(s):  
Filtration Rate ◽  
Distal Tubule ◽  
Inhibitory Effect ◽  
Proximal Convoluted Tubule ◽  
Proximal Tubules ◽  
Fluid Delivery ◽  
Collecting Ducts ◽  
Single Nephron ◽  
Distal Tubules ◽  
The Difference

1. Hydrochlorothiazide was administered at two doses to Long-Evans rats for 7–10 days. Both doses resulted in an initial natriuresis and diuresis. After 1 day of treatment the natriuresis abated, but the diuresis persisted. 2. The mechanisms responsible for these chronic effects were investigated by performing clearance and micropuncture studies on all animals; collections were made from late proximal tubules and from early and late regions of distal tubules. 3. Values for total glomerular filtration rate and single-nephron filtration rate in thiazide-treated rats were not significantly different from those in control animals. 4. The delivery of sodium to the end of the proximal convoluted tubule was considerably reduced in each group of thiazide-treated rats. Although sodium delivery to the early distal tubule was also significantly lower than in control animals, the difference had disappeared by the late distal tubule. 5. It is concluded that the return of sodium excretion to control levels during chronic hyrochlorothiazide administration is a consequence of increased fractional reabsorption by the proximal tubules, secondary to a thiazide-induced sodium depletion. This results in less sodium being delivered to the nephron site at which thiazides exert their major inhibitory effect. 6. Fluid delivery to the end of the proximal convoluted tubule and to the early distal tubule was significantly reduced in thiazide-treated rats; in animals given the higher dose of diuretic it was also significantly reduced at the end of the distal tubule. Nevertheless, in both thiazide-treated groups urine flow rate was elevated, suggesting that reabsorption of water from the collecting ducts is reduced during chronic thiazide administration.

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