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Dromedary camels can survive and reproduce in desert areas. The unique anatomical structure of the kidney enables the camel to prevent water loss. The present study aimed to investigate the ultrastructure of the peroxisomes in the normal kidney of the adult dromedary camel. Tissue samples were taken from the cortex and outer medulla of the kidney of eight camels. The samples were then processed for histological and ultrastructural investigations. The epithelial cells of the proximal tubules displayed peroxisomes with varying sizes and shapes. The peroxisomes were observed in either dispersed or clustered arrangement. Each peroxisome exhibited a homogenous matrix enveloped by a single membrane. Several peroxisomes exhibited one or more dark marginal plates that were always strongly associated with the smooth endoplasmic reticulum. The intensity of the peroxisomal matrix differed significantly, either within the same cell or across different cells. The intensity was light or dark, with a few peroxisomes presenting a similar intensity to that of the mitochondria. Some peroxisomes contained nucleoids within their matrix. The peroxisomes in the first and second sections of proximal convoluted tubules were scattered and primarily located in the region between the microvilli and the underlying mitochondria. The peroxisomes in the third region were abundant and frequently aggregated in clusters throughout the cytoplasm. In the fourth region, the number of peroxisomes was low. The proximal straight tubule had a limited quantity of peroxisomes. In conclusion, peroxisomes in the proximal tubule in kidney of normal dromedary camel were similar in shape and size to other mammals; however, heterogeneity exists as a result of differences in species-specific peroxisomal proteins. Peroxisomes are suggested to be a major source of metabolic energy and act as hydrogen peroxide (H 2 O 2 ) scavengers, resulting in the release of water and oxygen., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Ca 2+ is an important second messenger that translates extracellular stimuli into intracellular responses. Although there has been significant progress in understanding Ca 2+ dynamics in organs such as the brain, the nature of Ca 2+ signals in the kidney is still poorly understood. Here, we show that by using a genetically expressed highly sensitive reporter (GCaMP6s), it is possible to perform imaging of Ca 2+ signals at high resolution in the mouse kidney in vivo. Moreover, by applying machine learning-based automated analysis using a Ca 2+ -independent signal, quantitative data can be extracted in an unbiased manner. By projecting the resulting data onto the structure of the kidney, we show that different tubular segments display highly distinct spatiotemporal patterns of Ca 2+ signals. Furthermore, we provide evidence that Ca 2+ activity in the proximal tubule decreases with increasing distance from the glomerulus. Finally, we demonstrate that substantial changes in intracellular Ca 2+ can be detected in proximal tubules in a cisplatin model of acute kidney injury, which can be linked to alterations in cell structure and transport function. In summary, we describe a powerful new tool to investigate how single cell behavior is integrated with whole organ structure and function and how it is altered in disease states relevant to humans.

Background: The kidney proximal convoluted tubule (PCT) reabsorbs filtered macromolecules via receptor-mediated endocytosis (RME) or nonspecific fluid phase endocytosis (FPE); endocytosis is also an entry route for disease-causing toxins. PCT cells express the protein ligand receptor megalin and have a highly developed endolysosomal system (ELS). Two PCT segments (S1 and S2) display subtle differences in cellular ultrastructure; whether these translate into differences in endocytotic function has been unknown., Methods: To investigate potential differences in endocytic function in S1 and S2, we quantified ELS protein expression in mouse kidney PCTs using real-time quantitative polymerase chain reaction and immunostaining. We also used multiphoton microscopy to visualize uptake of fluorescently labeled ligands in both living animals and tissue cleared using a modified CLARITY approach., Results: Uptake of proteins by RME occurs almost exclusively in S1. In contrast, dextran uptake by FPE takes place in both S1 and S2, suggesting that RME and FPE are discrete processes. Expression of key ELS proteins, but not megalin, showed a bimodal distribution; levels were far higher in S1, where intracellular distribution was also more polarized. Tissue clearing permitted imaging of ligand uptake at single-organelle resolution in large sections of kidney cortex. Analysis of segmented tubules confirmed that, compared with protein uptake, dextran uptake occurred over a much greater length of the PCT, although individual PCTs show marked heterogeneity in solute uptake length and three-dimensional morphology., Conclusions: Striking axial differences in ligand uptake and ELS function exist along the PCT, independent of megalin expression. These differences have important implications for understanding topographic patterns of kidney diseases and the origins of proteinuria., (Copyright © 2018 by the American Society of Nephrology.)

The kidneys in two viviparous species of Neotropical lizards, Notomabuya frenata and Aspronema dorsivittatum (Mabuyidae), were investigated by light and scanning electron microscopy in order to determine the presence of the sexual segment of the kidney (SSK) and to study its morphology. The individuals used in this study belong to the Herpetological Collection of the Herpetology Laboratory - Reptiles of the Federal University of Juiz de Fora (CHUFJF-Reptiles) and they were collected between the years 2008 and 2012 from the Cerrado region in the state of Minas Gerais, Brazil. The SSK was present only in sexually mature males (with sperm in the testes / epididymis), whereas it was absent in sexually immature males. The nephron in both species consists of renal corpuscle, proximal convoluted tubule, distal convoluted tubule, collecting duct and sexual segment of the kidney. The SSK of the analyzed species were coated with a simple columnar epithelium, with high cells, basal nucleus and in the apical portion innumerable secretory granules. This study adds to the knowledge on reproductive biology and structures related to reproductive strategies of both lizard species and viviparous Neotropical lizards.

Parkinson's disease (PD) is a progressive neurodegenerative disorder for which there is no existing therapeutic approach to delay or stop progression. Genetic, biochemical and pre-clinical studies have provided evidence that leucine-rich-repeat-kinase-2 (LRRK2) kinase is involved in the pathogenesis of PD, and small molecule LRRK2 inhibitors represent a novel potential therapeutic approach. However, potentially adverse target-related effects have been discovered in the lung and kidneys of LRRK2 knock-out (ko) mice and rats. It is unclear if the LRRK2 ko effect in the kidneys and lung is also induced by pharmacological inhibition of the LRRK2 kinase. Here, we show that treatment with the LRRK2 inhibitor PFE-360 in rats induces a morphological kidney phenotype resembling that of the LRRK2 ko rats, whereas no effects were observed in the lung. The PFE-360 treatment induced morphological changes characterised by darkened kidneys and progressive accumulation of hyaline droplets in the renal proximal tubular epithelium. However, no histopathological evidence of renal tubular injury or changes in the blood and urine parameters that would be indicative of kidney toxicity or impaired kidney function were observed after up to 12 weeks of treatment. Morphological changes were detected in the kidney after 2 weeks of treatment and were partially reversible within a 30 day treatment-free period. Our findings suggest that pharmacological LRRK2 inhibition may not have adverse consequences for kidney function., (Copyright © 2018 H. Lundbeck A/S. Published by Elsevier B.V. All rights reserved.)

The kidney is the most important excretory organ in the body and plays an essential role in maintaining homeostasis in vivo by conserving body fluid and electrolytes and removing metabolic waste. In this study, three types of transgenic system were used to investigate the expression of the bitter taste receptor Tas2r105 in mouse renal tissue (Tas2r105-GFP/Cre, Tas2r105-GFP/Cre-DTA and Tas2r105-GFP/Cre-LacZ). The results suggest that bitter taste receptors Tas2r105 and Tas2r106 are expressed in the renal corpuscle and the renal tubule, including the proximal tubule and distal tubule. Expression of α-gustducin, an important component of taste signal transduction, was also detected in mouse kidney. Meanwhile, conditional diphtheria toxin (DTA) expression in Tas2r105+ cells caused an increase in size of the glomerulus and renal tubule, accompanied by a decrease in cell density in the glomerulus. This indicates that Tas2r105+ cells play an important role in maintaining the structure of the glomerulus and renal tubules. Overall, the current study collectively demonstrates that cells labeled by bitter taste receptor expression may play a critical role in controlling human health, and have properties far beyond the original concept of taste perception., (Copyright © 2015 Elsevier Inc. All rights reserved.)

We report the development of a small fluorescent molecule, BDNCA3-D2, herein referred to as PT-Yellow. Soaking zebrafish embryos in PT-Yellow or intraperitoneal injection into adults results in non-toxic in vivo fluorescent labeling of the renal proximal tubules, the major site of blood filtrate reabsorption and a common target of injury in acute kidney injury. We demonstrate the applicability of this new compound as a rapid and simple readout for zebrafish kidney filtration and proximal tubule reabsorption function.

MicroRNAs (miRNAs) are major posttranscriptional regulators of a wide variety of biological processes. However, redundancy among most miRNAs has made it difficult to identify their in vivo functions. We previously demonstrated that global inhibition of miRNA biogenesis in Xenopus resulted in a dramatically smaller pronephric kidney. This suggested that microRNAs play a pivotal role in organ size control. Here we now provide a detailed mechanistic explanation for this phenotype. We identified that the activation of the mechanistic target of rapamycin complex 1 (mTORC1) by Insulin and insulin-like growth factor (Igf) 2 is an important regulator in kidney growth, which in turn is modulated by microRNAs. Molecular analyses demonstrate that microRNAs set a threshold for mTORC1 signaling by down-regulating one of its core negative regulators, tuberous sclerosis 1 (Tsc1). Most importantly, this rheostat can be reprogrammed experimentally. Whereas knockdown of miRNAs causes growth arrest, concomitant knockdown of Tsc1 restores mTORC1 activity and proximal tubular size. Together, these data establish a previously unidentified in vivo paradigm for the importance of posttranscriptional regulation in organ size control.

D-Aspartate (D-Asp) is an endogenous substance in mammals. Degradation of D-Asp is carried out only by D-aspartate oxidase (DDO). We measured DDO activity in porcine tissues, and produced an anti-porcine DDO antibody to examine the cellular localization of DDO. All the tissues examined showed DDO activities, whereas the substrate D-Asp was not detected in kidney cortex, liver, heart, and gastric mucosa. In the kidney, intensive immunohistochemical staining for DDO was found in the epithelial cells of the proximal tubules. In the liver, the epithelial cells of interlobular bile ducts, liver sinusoid-lining cells with cytoplasmic processes, and the smooth muscle cells of arterioles were strongly stained for DDO. In the heart, cardiomyocytes and the smooth muscle cells of arterioles showed DDO-immunoreactivity. In the gastric mucosa, only the chief cells were DDO-positive. These newly identified DDO-positive cells seem to actively degrade D-Asp to prevent an excess of D-Asp from exerting harmful effects on the respective functions of porcine tissues.

This study details the gross and microscopic anatomy of the pelvic kidney in male Ambystoma maculatum. The nephron of male Ambystoma maculatum is divided into six distinct regions leading sequentially away from a renal corpuscle: (1) neck segment, which communicates with the coelomic cavity via a ventrally positioned pleuroperitoneal funnel, (2) proximal tubule, (3) intermediate segment, (4) distal tubule, (5) collecting tubule, and (6) collecting duct. The proximal tubule is divided into a vacuolated proximal region and a distal lysosomic region. The basal plasma membrane is modified into intertwining microvillus lamellae. The epithelium of the distal tubule varies little along its length and is demarcated by columns of mitochondria with their long axes oriented perpendicular to the basal lamina. The distal tubule possesses highly interdigitating microvillus lamellae from the lateral membranes and pronounced foot processes of the basal membrane that are not intertwined, but perpendicular to the basal lamina. The collecting tubule is lined by an epithelium with dark and light cells. Light cells are similar to those observed in the distal tuble except with less mitochondria and microvillus lamellae of the lateral and basal plasma membrane. Dark cells possess dark euchromatic nuclei and are filled with numerous small mitochondria. The epithelium of the neck segment, pleuroperitoneal funnel, and intermediate segment is composed entirely of ciliated cells with cilia protruding from only the central portion of the apical plasma membrane. The collecting duct is lined by a highly secretory epithelium that produces numerous membrane bound granules that stain positively for neutral carbohydrates and proteins. Apically positioned ciliated cells are intercalated between secretory cells. The collecting ducts anastomose caudally and unite with the Wolffian duct via a common collecting duct. The Wolffian duct is secretory, but not to the extent of the collecting duct, synthesizes neutral carbohydrates and proteins, and is also lined by apical ciliated cells intercalated between secretory cells. Although functional aspects associated with the morphological variation along the length of the proximal portions of the nephron have been investigated, the role of a highly secretory collecting duct has not. Historical data that implicated secretory activity concordant with mating activity, and similarity of structure and chemistry to sexual segments of the kidneys in other vertebrates, lead us to believe that the collecting duct functions as a secondary sexual organ in Ambystoma maculatum., (© 2010 Wiley-Liss, Inc.)

The three-dimensional architecture of nephrons in situ and their interrelationship with other nephrons are difficult to visualize by microscopic methods. The present study uses microcomputed X-ray tomography (micro-CT) to visualize intact nephrons in situ. Rat kidneys were perfusion-fixed with buffered formalin and their vasculature was subsequently perfused with radiopaque silicone. Cortical tissue was stained en bloc with osmium tetroxide, embedded in plastic, scanned, and reconstructed at voxel resolutions of 6, 2, and 1 microm. At 6 microm resolution, large blood vessels and glomeruli could be visualized but nephrons and their lumens were small and difficult to visualize. Optimal images were obtained using a synchrotron radiation source at 2 microm resolution where nephron components could be identified, correlated with histological sections, and traced. Proximal tubules had large diameters and opaque walls, whereas distal tubules, connecting tubules, and collecting ducts had smaller diameters and less opaque walls. Blood vessels could be distinguished from nephrons by the luminal presence of radiopaque silicone. Proximal tubules were three times longer than distal tubules. Proximal and distal tubules were tightly coiled in the outer cortex but were loosely coiled in the middle and inner cortex. The connecting tubules had the narrowest diameters of the tubules and converged to form arcades that paralleled the radial vessels as they extended to the outer cortex. These results illustrate a potential use of micro-CT to obtain three-dimensional information about nephron architecture and nephron interrelationships, which could be useful in evaluating experimental tubular hypertrophy, atrophy, and necrosis.

Aim: The renal tubular uptake of 125I-Aprotinin (*Ap) is on average located more superficially than its filtration site, causing transfer of some of *Ap filtered in deep to more superficial cortical zones. 125I-Cystatin C (*Cy) showed less uptake in deep cortical zones than Ap, suggesting a longer and/or a more superficial tubular uptake site. To test that hypothesis and to quantify the outward transfer of the filtered polypeptides, we estimated the tubular uptake pattern of the tracers in perfusion fixed rat kidneys after intravenous injection of *Cy and *Ap., Methods: Autoradiographs were made from 10 mum thick slices of Microfil nephron casts from outer (OC), middle (MC) and inner (IC) cortical zones to quantify cortical border-crossing *Ap transfer. Single nephron glomerular filtration rate (snGFR) was estimated as the zonal uptake of *Ap corrected for *Ap transfer, divided by its time-integrated plasma concentration and the zonal number of glomeruli., Results: *Ap and *Cy uptake fell exponentially along the proximal convoluted tubule (PCT), indicating an uptake proportional to luminal concentration. Uptake in IC exceeded that in MC and OC nephrons. The per cent PCT length with *Cy uptake (67.2 +/- 1.6) exceeded that of *Ap (54.6 +/- 1.8). The zonal border-crossing PCT length (29-34% of total PCT) from deep to more superficial cortical zones transferred 4-6% more *Cy than *Ap., Conclusion: Greater tubular uptake length of *Cy than of *Ap causes more cortical border-crossing of *Cy. The zonal snGFR estimated from Aprotinin uptake corrected for border-crossing agreed well with that obtained with the Hanssen ferrocyanide technique.

Background: Proximal tubule (PT) angiotensinogen (AGT) is part of a tubular renin-angiotensin system (RAS) that participates in the regulation of sodium reabsorption along the entire nephron. Physiologic maneuvers affecting AGT expression in PT also affect systemic RAS. Here, we tested the hypothesis that PT AGT is regulated by increased glomerular filtration rate (GFR)., Methods: Complete unilateral nephrectomy (UNX) in mice was used to induce a sustained increase in GFR in the remaining kidney. AGT expression was monitored by quantitative reverse transcription-polymerase chain reaction (RT-PCR). AGT protein in PT was investigated by semiquantitative histology. We also measured AGT concentration in plasma and in 24-hour urine by a specific enzyme-linked immunosorbent assay (ELISA)., Results: Seven weeks after nephrectomy, UNX animals exhibited a 2-fold increase in tubular AGT mRNA (P <.001) compared with sham-operated control animals. The proportion of PT sections exhibiting AGT immunostaining was significantly increased at day 3 (P <.05), and remained elevated at seven weeks (UNX = 0.63 +/- 0.09, sham = 0.38 +/- 0.02, P <.01), revealing recruitment of AGT-producing cells along the PT. AGT excretion in final urine corrected for creatinine and kidney weight was also elevated by UNX at seven weeks (UNX = 209 +/- 42 pmol/mg/g, sham = 147 +/- 29 pmol/mg/g, P <.05), with no difference in plasma AGT between UNX and control animals., Conclusion: These observations suggest that AGT expression in PT adapts in the long-term to changes in GFR. In the UNX model, urinary AGT excretion is also elevated as a consequence of increase in net tubular flow.

The DBA/2Cr mouse is characterized by the presence of giant lysosomes located in the proximal convoluted tubules of males and proximal straight tubules of females. However, it remains unclear whether these giant lysosomes in the proximal tubules are characteristic of DBA/2Cr specifically, or are common to other DBA/2 substrains and DBA/1. The present study investigated the morphology of kidneys from DBA/2CrSlc, DBA/2JJcl, DBA/2NCrj and DBA/1JNCrj mice of both sexes. Giant lysosomes in the renal proximal tubules were found to represent common morphological characteristic of both DBA/2 and DBA/1JN.

The review presents a modern view on the functional--morphological peculiarities of 12 segments of the nephron canaliculi possessing some functional, biochemical and histologic distinctions that should be taken into account to study physiology, biochemistry, morphology and pathophysiology of the kidney.

In the present study, we performed comparative histological observations of ICR, BALB/c, C57BL/6, C3H/HeN and DBA/2 mice kidneys. Sex and strain differences were observed in the appearance of vacuolar structures of the proximal convoluted tubules (toluidine blue-positive granules in osmium-postfixed epoxy-resin sections). These features were especially remarkable in male DBA/2 mice. The vacuolar structures in male DBA/2 mice showed heterogeneous staining with Sudan B in frozen sections and appeared under an electron microscope as multilammelar giant dense bodies. In addition, these dense bodies showed heterogeneous acid phosphatase reactions. Immunohistochemical analyses of these structures for apolipoprotein B showed strong positive reactions. These results suggested that vacuolar structures in the proximal convoluted tubules, which were remarkable in male DBA/2 mice, were giant lysosomes containing apolipoprotein B.

We developed a model of volume contraction in rabbits by using a furosemide/low-salt diet to follow changes, if any, in proximal tubule Na+/H+ exchanger 3 (NHE3) mRNA and brush-border protein. The rabbits' plasma renin, aldosterone, and urine sodium content confirmed the volume-contracted state. RNase protection assays demonstrated increases in treated-animal NHE3 mRNA as a percentage of control with 172 +/- 23, 154 +/- 15, 153 +/- 14, and 141 +/- 7 (SE) % (P < 0.05) at 1, 5, 10, and 31 days, respectively. Western analysis of brush-border membrane with NHE3 antibody revealed increased immunoreactivity in treated animals as a percentage of control with 120 +/- 30, 190 +/- 59, 307 +/- 72, and 427 +/- 41% (P < 0.05) at 1, 5, 10, and 31 days, respectively. There was no significant difference in serum potassium, bicarbonate, and cortisol in control vs. experimental animals. These data suggest that there is chronic upregulation of NHE3 in the volume-contracted state.

Seasonal variability in kidney morphology of the house sparrow Passer domesticus was examined using light microscopy. Sparrows were captured from the wild in winter, spring, summer and autumn. The kidneys were perfused with half-strength Karnovsky's fixative and processed for light microscopy by embedding in either paraffin wax or JB4 acrylic resin. Absolute volumes of the kidneys, their components (cortex, medulla and blood vessels), components of the nephron (renal corpuscles, proximal tubules, loops of Henle, distal tubules and collecting ducts) and the capillaries surrounding the nephron were quantified using stereology. Tissue processed in paraffin wax had a mean shrinkage of 17.7 % compared with 10.1 % in JB4 resin. Absolute volumes of the kidneys and nephrons were compared statistically between tissue processing methods and among seasons. Absolute volumes of the structures within the kidneys were not significantly different between treatments or among seasons. Within the nephron, the only measured variables to show significant differences were the absolute volumes of the distal tubules and cortical collecting ducts between tissue processing treatments. Thus, kidney morphology was relatively unaffected by changes in season. In addition, the results show that embedding tissue using acrylic resin causes less shrinkage, and it should therefore be the preferred embedding medium for quantitative morphologists.

The organization of the renal medulla of the Gambel's quail, Callipepla gambelii, kidney was examined to determine the number of loops of Henle and collecting ducts and the surface area occupied by the different nephron segments as a function of distance down the medullary cones. Eleven medullary cones were dissected from the kidneys of four birds, and the tissue was processed and sectioned for light microscopy. In addition, individual nephrons were isolated on which total loop thin descending segment and thick prebend segment lengths were measured. The results show no correlation between the absolute number of loops of Henle and the length of the medullary cones. The number of thick and thin limbs of Henle and collecting ducts decrease exponentially with distance toward the apex of the cones and the rate of decrease is similar for cones of different lengths. Initially there is a rapid decrease in the number of thin limbs of Henle, indicating that most nephrons do not penetrate the cones a great distance. Thick descending limbs of Henle (prebend segment) ranged in length from 50 to 770 microm, and there was little correlation with the total length of the loop of Henle. However, the length of the thin limb of Henle correlated well with total loop length. The cell surface areas of the limbs of the loop of Henle and the collecting ducts decreased toward the apex of the cones.

Glutamine synthesis, a major process for ammonia detoxification and the control of acid-base balance, occurs from various precursors in suspensions of rabbit proximal tubules. However, no data are currently available on the distribution of glutamine synthesis along the rabbit proximal tubule, and its modulation by changes of substrate concentration. Therefore we have microdissected and incubated the three parts (S1, S2 and S3) of rabbit proximal tubules and measured glutamine synthesis from alanine and aspartate. With a physiological concentration of alanine (0.25 mM) or aspartate (0.05 mM), glutamine synthesis in the S1 segment was about half of that in the S2 and S3 segments, and was greater from alanine than from aspartate along the entire proximal tubule. Elevation of alanine and aspartate concentrations to 5 mM increased glutamine synthesis in both a substrate- and segment-dependent manner. It is concluded that glutamine synthesis occurs from alanine and aspartate along the entire rabbit proximal tubule; however, contrary to what might have been expected on the basis of measurement of glutamine synthetase activity, the basal rate of glutamine synthesis and its adaptation to increased substrate availability are heterogeneous along this nephron segment.

Diabetes in now the commonest cause of renal failure in the western world. Furthermore the survival of diabetic patients requiring dialysis treatment for renal failure is far less than patients with renal failure secondary to all other diseases. It is therefore important to identify the factors that control the development of progressive renal disease to allow targeted therapeutic interventions which would have major implications both to patient well-being and also to the provision of health care world wide. In this review we discuss possible metabolic consequences of hyperglycemia and their role in the pathogenesis of diabetic nephropathy. We also focus on the involvement of the pro-fibrotic cytokine Transforming Growth Factor beta, and contrast its role in the pathogenesis of glomerular and tubulo-interstitial changes seen in diabetic nephropathy.

Effects of maternal uninephrectomy in rats, which was performed on day 5 of gestation, on development of proximal renal tubule of the pups were morphometrically investigated. One day after birth, both the proximal tubular length per unit volume (1 mm3) of whole kidney and total proximal tubular volume in the neonates from uninephrectomized mothers were significantly larger than those in the neonates from sham-operated mothers. Six weeks after birth, no significant differences in parameters of the kidney were observed between the pups from uninephrectomized and sham-operated mothers. These results suggest that maternal uninephrectomy in the rat accelerates the development of proximal renal tubule in the fetus and that the renotropic activity is not lasting during postnatal development of the kidney.

The subcellular distribution and segmental variations in location of gp330, a scavenger receptor for filtered proteins in renal proximal tubules, was analyzed. Kidney tissue from rats (4 different strains), rabbits and humans were analyzed by light- and electron microscope immunocytochemistry, using cryosections or Lowicryl sections from cryosubstituted tissue. Gp330 was located mainly in apical coated pits, small and large endocytic vacuoles and in dense apical tubules in the proximal tubule cells. The labeling density was markedly higher in segments 1 and 2 as compared to segment 3 of the proximal tubule. In addition to the location in the early part of the endocytic pathway, gp330 was also present in lysosomes, especially in segments 1 and 2. The lysosomal labeling was not restricted to the membrane, but was also seen in the matrix. Localization of gp330 in lysosomes was confirmed on sections from purified lysosomal fractions from rat renal cortex. The brush border localization of gp330 in proximal tubules exhibited a characteristic segmental variation. In the initial part of segment 1, there was virtually no brush border labeling. In the remaining part of segment 1 and in segment 2, there was a distinct but sometimes patchy labeling of the brush border. In segment 3, groups of microvilli of approximately 10 as seen in sections were intensively labeled from bottom to tip and there were often more than one of these groups on a single cell, the remaining microvilli were unlabeled. No differences in the cellular and subcellular localization of gp330 were observed between species or rat strains. In conclusion, the present study demonstrates that in addition to its location in the early endocytic and recycling pathway, gp330 is also present in microvilli and the protein and degradation products thereof is present in lysosomes, consistent with its role as a protein scavenger receptor.

Previous research in animals and humans has demonstrated that many nephrotoxic chemicals induce selective injury within the kidney affecting either renal proximal straight (PST) or proximal convoluted (PCT) tubules. Selective injury has also been observed following in vitro nephrotoxicant exposure to precision-cut renal slices and isolated PCT and PST segments. These in vitro models provide a means of comparing and contrasting basic mechanistic differences which render these segments innately susceptible to nephrotoxicant injury. In this article, methods for preparing precision-cut slices and isolating PST and PCT segments will be reviewed.

Oligonucleotide probes were designed to sequences of the rat alpha 1B- and alpha 2B-adrenergic receptor mRNA and used for in situ hybridization histochemistry on tissue sections of kidneys from control and thyroidectomized rats. Both alpha 1B- and alpha 2B-receptor mRNA labelling was demonstrated in proximal tubule cells in the outer stripe of the outer medulla, with tubular rays radiating into the cortex. Thyroidectomy induced a more than 4-fold increase in mRNA for the alpha 1B-receptor in the kidney, whereas no change in alpha 2B-receptor mRNA levels could be demonstrated in thyroidectomized rats as compared to control animals. The results suggest that thyroid hormone plays an important role in regulating expression of alpha 1B-receptors in renal tubule cells.

The ultrastructure of renal tubule cells was studied in the European lesser spotted dogfish by the evaluation of thin sections and freeze fracture replicas. Computer-assisted three-dimensional reconstruction of entire nephrons was performed. The distinction of nephron segments and collecting tubule was made using results of previous histological work. The first proximal tubule segment (PI) consists of two subsequent portions, PIa and PIb. PIa is a component of the lateral countercurrent bundle, and PIb, which displays an apical tubulovesicular apparatus and an extended lysosomal compartment, is located in the vicinity of the glomeruli. Rod-shaped intramembrane particles were detected in PIa. The second proximal tubule segment (PII) is a special segment in elasmobranch and teleost fish. PII differs largely from PI in cell morphology and function. The apical cytoplasm was filled with small clear vesicles, and an apical endocytic apparatus was lacking. In the apical cell membrane, rod-shaped particles were revealed by freeze fracture. The apical tight junctions of PI and PII consisted of seven to ten meandering strands. The distal nephron was subdivided into two major segments: early distal tubule (EDT) in the lateral countercurrent bundles and late distal tubule (LDT) in the mesial tissue. The EDT showed marked amplification of basolateral cell membranes. The tight junctions displayed a low number of continuous parallel strands, which is also characteristically found in the diluting segments of other vertebrates. LDT cells showed cytoplasmic studs and rod-shaped intramembraneous particles at the apical cell membrane, thereby resembling type A intercalated cells of collecting duct. The collecting tubule (CT) emerged from the LDT and was part of the countercurrent arrangement inside the lateral bundles. Tight junctions of LDT and CT consisted of many meandering strands in a honeycomb pattern. With immunohistochemistry, binding sites of a polyclonal antibody against an extraplasmic portion of rat gastric H(+)-K(+)-adenosine triphosphatase (ATPase) were observed at the apical cell membrane of PIa, PII, and LDT. From the colocalization of binding sites for the antibody against the transport enzyme with rod-shaped intramembrane particles, we assume that these might be the morphological correlate of gastric H(+)-K(+)-ATPase-like enzyme in the renal tubule.

The platypus kidney shows morphological similarities to those of other mammals. Macroscopically, the cortex is easily distinguishable from the fairly wide medulla. Within the medulla, no clear border is observed between the inner and outer zones. Light and transmission electron microscopically, the glomeruli show quite similar architecture to those of other mammals; however, the glomerular lobulation is very clear. The glomerular tufts are rather simple, but capillary lumen varies widely in size, which is one of the unique features of the platypus kidney. The urinary tubule is generally similar to that of human and other mammals in shape and segmentation; however, the staining specificities of histochemical reactions and the shape of epithelial cells of the Henle's loop differ from those of other mammals. The most conspicuous features are: 1) although no protein casts are found in the tubular lumina, epithelial cells of the proximal convoluted tubule (PCT) have numerous electron-dense vesicles as in human nephrotic kidneys; and 2) the platypus Henle's loop consists of the thick epithelial cells similar to the mammalian type nephron of birds. As compared to those of other mammals such as humans and rats, our observations suggest that the platypus kidney is less developed, in terms of evolution.

120 male rats belonging to two different strains (Han: WIST, Han: SPRD) and two different weight groups (group I: lighter body weight, group II heavier body weight) were killed on three different methods (decapitation, Nembutal -overdose greater than 250 mg/kg b.w. i.p. less than, exsanguination in Nembutal -anaesthesia greater than 100 mg/kg b.w. i.p.). Subsequently, changes in tubuli contorti I of the left kidneys were examined morphometrically. The mean total area, luminal area and epithelial area were evaluated as well as the mean diameter of the tubular nuclei. Concerning the areas of the proximal tubules only the luminal areas show differences between the Wistar and the Sprague-Dawley strain. The total, luminal and epithelial area in the rats belonging to the weight group II were significantly larger than in the rats belonging to weight group I. With one exception this could be noticed within both strains and all three modes of killing. In both strains and weight groups the rats were killed by Nembutal -overdose or exsanguination showed significantly smaller total, luminal and epithelial areas than the rats that were decapitated. Possible cause heretofore are discussed. Differences between the tubular areas of the rats that were killed by Nembutal -overdose and those that were killed by exsanguination after Nembutal -anaesthesia could not be established. The measurements of nuclei diameters in the proximal tubules did not lead to clearly different results between both strains, weight groups and the various modes of killing.

Using the cheek pouch of the Syrian hamster as an experimental model for oral carcinogenesis, it has been shown that the expression of transforming growth factor-alpha (TGF-alpha) is consistently associated with the malignant transformation process. We have recently shown that production of TGF-alpha has been localized to normal hamster oral epithelium and bone marrow eosinophils. In this study we investigated the production of this cytokine in other normal hamster adult tissues. By using an EGF-radioreceptor assay, immunohistochemistry, Northern blot analysis, and in situ hybridization we have now further detected the presence of TGF-alpha mRNA and/or protein in the kidney, stomach, and pancreas of normal adult hamster. Together with the previously reported detection of TGF-alpha in oral mucosa and bone marrow eosinophils, these adult normal tissue/cellular sources can serve as sites of TGF-alpha production. The availability of hamster specific reagents (cDNA and monoclonal antibodies) and the delineation of the various adult tissues that could produce TGF-alpha make the Syrian hamster a suitable model for the study of how this multifunctional cytokine can influence normal and pathological processes.

In dual micropuncture experiments the shrinking drop technique was used to measure volume flux with artificial tubular fluid (AF) alternating with harvested tubular fluid (HTF) during saline volume-expanded (VE) and nonexpanded (NE) periods. In VE rats, volume flux (Jv) (nl.mm-1.min-1) with AF was 1.78 +/- 0.08 (means +/- SE) during NE and was reduced to 1.39 +/- 0.09 (P = 0.01) during subsequent VE, whereas with randomly alternating HTF during VE it was 1.07 +/- 0.08 (P less than 0.0001 and less than 0.03, respectively). Jv with HTF from NE rats tested in the VE rats was 1.20 +/- 0.06, which is significantly higher than that measured with their own HTF (Wilcoxon rank, P = 0.05). In NE rats Jv was 1.65 +/- 0.10 and 1.69 +/- 0.10 with AF and HTF and was 1.28 +/- 0.07 (P less than 0.001 from both) with HTF obtained from VE rats. Elemental analysis of reaspirated tubular fluids showed no significant differences in Na or Cl concentrations among any of the fluids. It is concluded that during VE, a transferable Na transport inhibitor appears in proximal tubular fluid, which, together with a changed proximal tubular epithelium, possibly due to physical forces, accounts for proximal tubular Na transport inhibition during VE.

The structure of the nephron in the ranid frog, Rana cancrivora, was studied by light and electron microscopy. This frog is the only amphibian species to live in mangrove swamps of very high salinity. The nephron consists of the following parts: renal corpuscle, ciliated neck segment, proximal tubule, ciliated intermediate segment, distal tubule, connecting tubule, and collecting duct. The distal tubule is located in the ventromedial region of the kidney, and the other tubules are situated in the dorsolateral region. Renal corpuscles are found between the two regions. Some renal corpuscles have a wide Bowman's space because of the small glomerulus within them. The proximal tubules are composed of columnar cells with a dense luminal brush border of long microvilli and numerous apical vesicles and vacuoles. The initial part of the distal tubule consists of heavily interdigitated cells, characterized by a very regular palisade arrangement of mitochondria. In the terminal part of the distal tubule, shorter mitochondria of the infolding cells are situated irregularly around the nucleus. The connecting tubule consists of principal cells and canaliculus cells. The collecting duct consists of columnar or cuboidal cells; cytoplasmic organelles are relatively sparse. The canaliculus cells are intercalated between principal cells from the terminal distal tubule to the proximal part of the collecting duct. Our findings indicate that the kidney of R. cancrivora is structurally similar to kidneys of other amphibians. These findings are discussed with regard to probable correlations between ultrastructure and function in R. cancrivora.

The literature on the role of the kidney and renal morphological modifications in places of limited water supply is reviewed. The anatomical structures for urine concentration found in animals living in desert or arid environments, although not all occurring in one particular animal, are wide medullae, long loops of Henle, long proximal tubules, long collecting tubules, small renal corpuscles, extension of the renal pelvis, well developed elongated papillae, occurrence of giant vascular bundles, specialized ultrastructure of Henle's loops, epithelial changes in the collecting tubule, zonation of the vasa recta and peculiarity of the arterial supply to the kidney. The renal renin content is moderately high in these species. The renin-angiotensin-aldosterone system is very active, retaining Na+ with water. The urine is concentrated at the expense of other electrolytes. Both the renal blood and urinary flow rates are lower than in species with access to unlimited water supply. The juxtaglomerular apparatus components are topographically intimate for effective tubuloglomerular autoregulation of renal blood flow.

As demonstrate the experiments performed in 45 male rats of Wistar line, a complete bile loss by the organism results in 24-72 h in certain distrophic changes of epithelium in the proximal parts of the nephron canaliculi. The brush border microvilli undergo destruction, the cellular membranes disorganize, cytoplasm of epitheliocytes vacuolizes up to their complete rupture. The PAS-positive reaction of the basal membrane in the canaliculi increases. In cytoplasm of epitheliocytes in the straight portions of the canaliculi of the distal parts of the nephron glycogene is deposited. Alkaline phosphatase, succinate dehydrogenase, cytochromoxydase activity decreases, and that of acidic phosphatase increases. The changes advance with increasing time from the beginning of bile loss.

The technique of microdissection and the structure of nephrons and collecting ducts from normal kidneys were studied. The structure of the assumedly normal nephrons and collecting ducts is characterized by the uniformity of the outlines of the tubules. The various parts of the tubules are distinguished by the differences in diameter and epithelial structure.

Late proximal rat tubular segments were microperfused with slightly hypo- or hypertonic artificial late proximal tubular fluid (ATF) at low (11-13 nl/min) or high (30-38 nl/min) perfusion rates. Volume reabsorption, net chloride and solute reabsorption were measured as a function of length. In addition, the transepithelial resistance and voltage (Vte) were measured as a function of the applied osmotic gradient. Hypertonic solutions equilibrated to isotonicity by solute outflow rather than water influx. With hypertonic ATF the lumen positive Vte was decreased compared with free flow or with hypotonic ATF. The resistance was not significantly different between the different groups. In contrast to hypotonic ATF, hypertonic or isotonic ATF was not significantly reabsorbed. In addition, hypotonic ATF maintained its hypotonicity along the perfused segments. Its reabsorption was flow-dependent. Hypotonicity appeared to enhance solute reabsorption.

The single nephron glomerular filtration rate (SNGFR) was determined in superficial (S) and juxtamedullary (JM) nephrons of 10 anesthetized rabbits by the 14C ferrocyanide infusion technique. The length of the proximal tubules and the volume of the glomeruli were also determined for the same nephrons. SNGFR was higher in JM than in S: 28.6 +/- 3.4 versus 22.6 +/- 3.0 nl/min, P less than 0.001. In JM nephrons, glomeruli were larger than in S: 1.3 +/- 0.1 versus 0.9 +/- 0.1 nl, P less than 0.001, whereas there was no difference between proximal tubule length in either category (s, 8.7 +/- 0.3 and JM, 8.9 +/- 0.5 mm). In 6 out of 8 animals, SNGFR was significantly correlated to glomerular volume. Lack of correlation was observed between SNGFR and length of proximal tubule in all animals but one. These results show that the rabbit, as well as small rodents and the dog, has a higher SNGFR in juxtamedullary than superficial glomeruli. Although this functional difference is not related to the length of the proximal tubule in each individual animal, the ratio between the mean SNGFR value and the mean length of the proximal tubule in superficial rabbit nephrons is similar to the ratio found in other species.

1. In vivo micropuncture techniques, with and without peritubular capillary perfusion, were used to study the effects of high extracellular Na and Cl concentrations on transepithelial volume (Jv) and sodium (JNa) fluxes in rat proximal tubules. 2. In a double blind manner, the shrinking drop technique of Gertz was used to measure Jv; JNa was calculated from this and the tubular fluid Na concentration. 3. At both 184 and 279 mmol/l pericellular Na concentrations (both inside and outside the tubular epithelium), net Jv decreased significantly by 15 and 64%, respectively. Net JNa remained constant at 184 but decreased by 29% at 279 mmol/l Na concentration. 4. Thus, at both Na concentrations, when translated to free flow conditions, fractional Na reabsorption must have decreased. These findings, also supported by previous results at these Na concentrations, indicate that active Na transport was inhibited by high pericellular Na concentrations. 5. When intratubular Cl concentration was varied between 108 and 138 mmol/l while peritubular Cl was maintained constant (blood perfusing the capillaries), neither Jv nor JNa changed. Thus, at zero tubular flow, differential Cl/HCO3 concentrations do not provide significant driving forces for net Jv or JNa. 6. When only intratubular but not peritubular Na was elevated to 279 mmol/l, Jv and JNa increased markedly by 50 and 187%, providing evidence that a true solvent drag (solute drag) effect does exist in rat proximal tubules. 7. These findings offer a mechanism to explain why Na reabsorption is not increased when the filtered load of Na is increased with an elevation of plasma Na.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphologic and physiologic studies have established that filtered proteins are absorbed in the proximal tubule by endocytosis and transported to the lysosomes for degradation. The tubular absorption, hydrolysis and accumulation of albumin were examined in all three segments of the rabbit proximal tubule. S1, S2 and S3 segments were dissected and perfused in vitro with tritiated albumin at a physiologic concentration of 0.0364 mg/ml, and with [14C]inulin to determine fluid reabsorption. In addition, the three segments were fixed for ultrastructural examination after perfusion under conditions similar to those in the physiologic studies. The fluid reabsorption was similar in S1 and S2 but lower in S3. Albumin absorption was unexpectedly similar in the three segments. A lower percentage of absorbed albumin was hydrolyzed in the S3 segment compared with the earlier segments. The values were 70 +/- 15%, 61 +/- 11%, and 30 +/- 4% for S1, S2, and S3, respectively. The cellular accumulation of protein was highest in the S3 segment. The ultrastructure of the three segments was similar to that described in in vivo preserved kidneys, and no abnormalities were observed in the endocytic-lysosomal compartment. These results reveal axial heterogeneity in the hydrolysis of absorbed albumin by the rabbit proximal tubule and suggest that under normal physiologic conditions the S3 segment has a lower lysosomal proteolytic activity. Although the S3 segment maintains a high capacity for protein absorption, the earlier proximal segments likely have a greater role in protein degradation.

The present studies tested the hypothesis that osmotic disequilibrium between luminal and peritubular fluids is the driving force for net volume absorption in the isolated proximal straight tubule. Isolated tubule segments from superficial rabbit renal cortex were perfused at varying rates with a high chloride and bicarbonate-free solution as they were bathed with a normal bicarbonate-Krebs-Ringer buffer solution at 38 degrees C. Increasing the perfusion rate from congruent to 4 to congruent to 30 nl/min produced a monotonic increase in net volume absorption (Jv) from 0.18 +/- (sem) 0.03 to 0.62 +/- 0.08 nl . min-1. The chloride concentration in collected fluid samples rose from congruent to 137 to congruent to 147 mEq/liter over the same perfusion rate range. Ouabain (10(-4) m) added to the bathing solution inhibited Jv by a rate which varied from 0.20 to 0.28 nl . min-1 . min-1, depending on the perfusion rate. A mathematical model of the axial flows and transepithelial transport processes was developed. This model, and the experimental data, is consistent with the view that the driving force for isotonic fluid absorption in these tubules depends on the axial maintenance of osmotic disequilibrium between the perfusate and the bathing solution. Increasing the perfusion rate opposes osmotic equilibration by minimizing the extent to which dissipative fluxes of chloride and bicarbonate ions change the transepithelial chloride and bicarbonate concentration gradients, and by minimizing the tendency of the luminal cryoscopic osmolality to increase as volume absorption occurs.

This review contrasts the glucose transport mechanisms at opposing surfaces of the renal proximal convoluted tubule: the Na+-dependent D-glucose transporter localized at the brush border membrane and the Na+-independent transporter localized at the basolateral surface. The two sugar transport mechanisms are discussed from the point of view of their specificity, kinetic, and regulatory behaviors. Recent results focussing on molecular characterization of these different carrier proteins are also described, including some newer information on purification of the Na+-dependent glucose carrier from the brush border membrane.

Rabbits were anaesthetized and kidneys removed directly with no flush (group NF), or alternatively kidneys were flushed (group F) with sodium phosphate buffered 140 mmol/l sucrose (PBsuc 140), Collins C2 (C2) or Euro-Collins (EC) transplant preservation solutions and stored at 4 degrees C for 0-4, 24 or 48 h. Mid-cortical proximal convoluted tubule (PCT) segments were dissected from all groups and set up for microperfusion in vitro. Observations were made of tubule morphology, fluid reabsorption rate (Jv), bath leak of the glomerular marker iothalamate and transmural potential difference (p.d.), both at 37 degrees C and at 15-20 degrees C, in order to compare the relative effectiveness of the solutions in the preservation of tubule integrity and immediate function. Tubules from kidneys flushed with EC and C2 contained luminal debris and frequently had high bath leaks of iothalamate, both indicative of poor preservation of tubule integrity. In contrast, tubules from NF kidneys and PBsuc 140-flushed kidneys were free of luminal debris with a lower incidence of high iothalamate leaks. Tubules from PBsuc 140-flushed and EC-flushed kidneys after 0-4 and 24 h storage had Jv that were similar to those of NF kidneys. Tubules from all groups of flushed kidneys after 48 h cold storage and all tubules from C2-flushed kidneys showed reduced Jv values. In all cases Jv was reduced to approximately zero when the temperature was lowered to 15-20 degrees C. Transmural p.d. was similar in all groups except for particularly low p.d. values observed in tubules from C2-flushed kidneys after 48 h storage. These observations suggest that PBsuc 140 is more effective in the preservation of tubule function during prolonged cold storage than the glucose-based Collins solutions. The marked difference in the effectiveness of C2 and EC contraindicates the inclusion of magnesium (present in C2) for preservation of kidneys, as judged by experiments on the rabbit.

Proximal convoluted tubules (PCTs) from rabbit cortical slices were perfused after preservation in Collins' solution with the in vitro microperfusion technique. Under these conditions, after maneuvers of in vitro preservation, the following findings were observed: Tubules were best preserved, functionally and morphologically, when bathed with Collins' solution peritubularly. Tubular preservation was inadequate when the Collins' solution contacted only the luminal or luminal and peritubular sides. These observations indicate a difference in the reactions of various cellular sides to the preservation process, suggesting that during preservation of the whole kidney, there are differences in the preservation of the various tissues of the organ.

The nephron and collecting ducts of the little skate (Raja erinacea) and spiny dogfish shark (Squalus acanthias) have been investigated by light microscopy of semi-thin sections. Parts of the tubules (collecting ducts and distal segments) were identified after tubular injections with Microfil or carbon. The bundle zone was studied in serial sections. In the sinus zone transitions between the different segments were recorded. Thus, a complete reconstruction of the nephron, its subdivision into segments, and their localization in the kidney was accomplished. The nephron makes 4 loops. Beginning at Bowman's capsule, which sits between the bundle zone and sinus zone, the first loop is in the bundle zone. The nephron then extends into the sinus zone and turns back forming the second loop. This is followed by a third loop in the bundle zone which descends again into the sinus zone to form the last loop. The tail of the last loop (distal tubule) goes into the bundle zone and joins the collecting ducts. These collecting ducts are in the subcapsular connective tissue and progressively fuse to form a collecting tube. In the skate this tube traverses the thickness of the kidney between adjacent renal lobes to exit on the ventral kidney surface. In the shark, the large collecting ducts run on the surface of each lobe toward the medial margin of the kidney. Loops one and three and the early distal segment--all belonging to the same nephron--and a network of anastomosing capillaries form a bundle enclosed by a sheath of overlapping squamous cells termed "peritubular sheath." This anatomical unit forms the renal countercurrent system of the marine elasmobranch. The tubular bundle has a straight portion in which the nephron segments are arranged in a highly parallel fashion. The remainder of the bundle and of the encasing peritubular sheath are convoluted. The sequence of the tubule morphology beginning at Bowman's capsule is: neck segment (early and late), proximal tubule (four portions), intermediate (six portions), distal tubule (early and late), collecting duct (early and late).

The microscopic structure of the various nephron segments comprising the medullary counterflow system is described. The architectural organization of the tubules and blood vessels in the inner and outer medulla is also described. Species differences are noted. A hypothesis that the thicker parts of the descending limbs of Henle actively secrete salt is advanced.

Although each of the major experimental techniques applied to the study of renal physiology has provided its fair share of new information, the technique of in vitro microperfusion of nephron segments is notable for two major contributions. First, it has supplied a more direct and controlled means of studying epithelial transport processes, some of which already have helped us to understand certain aspects of kidney function and others of which have yet to find their application in unraveling the mysteries of the kidney. Second, in the process of delineating these transport characteristics, it has served to emphasize the epithelial specialization present in the kidney, providing functional counterparts to the already recognized anatomic heterogeneity present in the kidney. In this second role microperfusion has spawned the application of biochemical analysis of the hormonal responses of various nephron segments and contributed to the impetus for work in culturing the various cell types present in each nephron segment. This review outlines the functional characteristics of the 11 major segments of the nephron, incorporating what has been learned from some of the biochemical work on hormone response and correlating the latter with transport events.

ALP and gamma-GT are 2 brush border enzymes that can be individually demonstrated on adjacent sections by the histochemical methods of Mayahara (ALP) and Rutenberg (gamma-GT). On the basis of each enzyme activity, it was possible to recognize different categories of tubules in the mouse nephron. In fact, both enzymes were heterogeneously distributed along the proximal tubule, but in opposite gradients. The various staining intensities probably corresponded to proximal segmentation, but were sometimes difficult to evaluate. A technique was perfected to localize both enzymes in the same tissue section. Since each enzyme produced a distinct type of colored precipitates (ALP: black, gamma-GT: red), 4 categories of tubules could be identified, according to staining characteristics: 1. black tubules where ALP activity was predominant, corresponded to S1 segments, 2. black and red tubules where the 2 activities were about equivalent, were considered as parts of S2, 3. red ones where gamma-GT activity was high, were identified as portions of S3, 4. negative tubules where no activity was apparent, represented distal and straight collecting tubules. In addition to economize time and tissue, this simple technique permits to easily estimate variations in enzyme activities that probably correspond to structural and functional differences in the segments of the proximal tubule.