Morphometry: assessing direct and indirect methods of measuring the diameters of tubular structures / Morfometría: evaluación de métodos directos e indirectos para medir los diámetros de las estructuras tubulares

SUMMARY: Knowledge of the diameter of a structure or particle is required for stereological calculations. However, there is no consensus on the methodology for its measurement. This study aims to assess the differences between direct and indirect methods of measuring diameter. It is hypothesised that kidneys were removed, fixed, processed, sectioned, and stained. The stained slides were imaged using a digital microscope. The images were processed using the ImageJ software. The diameters of the renal glomeruli and collecting tubules were measured using direct and indirect methods. The measured diameters were analysed using the SPSS software v20. The differences between the measurements were assessed using a Z-test and test of association, and P < 0.05 was considered significant. No significant differences were observed between the diameters of the glomeruli (P = 0.82) and proximal (P = 0.86) and distal (P = 0.55) convoluted tubules as measured via direct and indirect methods. There was a strong positive correlation between the diameters of glomeruli (P = 0.97) and proximal (P = 0.82) and distal (P = 0.93) convoluted tubules measured using the two methods, both of which are convenient, accurate and suitable. The P-values based on these measurements were more than 0.05. Therefore, the study hypothesis was rejected. There was no significant difference between the direct and indirect methods of measuring diameter, and the null hypothesis was rejected; thus, both methods can be applied either independently or jointly.

RESUMEN: Se requiere el conocimiento del diámetro de una estructura o partícula para los cálculos estereológicos. Sin embargo, no existe consenso sobre la metodología para su medición. Este estudio tuvo como objetivo evaluar las diferencias entre los métodos directos e indirectos de medición del diámetro de una estructura. Riñones de ratas Wistar fueron extirpados, fijados, procesados y seccionados, y luego se tiñeron con HE. Se tomaron imágenes de las muestras teñidas usando un microscopio digital. Las imágenes fueron procesadas utilizando el software ImageJ. Los diámetros de los glomérulos renales y túbulos colectores se midieron por métodos directos e indirectos. Los diámetros medidos se analizaron utilizando el software SPSS v20. Las diferencias entre las medidas se evaluaron mediante una prueba Z y una prueba de asociación, y se consideró significativa P < 0,05. No se observaron diferencias significativas entre los diámetros de los glomérulos (P = 0,82) y túbulos contorneados proximales (P = 0,86) y distales (P = 0,55) medidos mediante métodos directos e indirectos. Hubo una fuerte correlación positiva entre los diámetros de los glomérulos (P = 0,97) y los túbulos contorneados proximales (P = 0,82) y distales (P = 0,93) medidos con los dos métodos, ambos convenientes, precisos y adecuados. Los valores P basados en estas mediciones fueron superiores a 0,05. Por lo tanto, se rechazó la hipótesis del estudio. No hubo diferencia significativa entre los métodos directo e indirecto de medición del diámetro, y se rechazó la hipótesis nula; por lo tanto, ambos métodos se pueden aplicar de forma independiente o conjunta.

Glomerular loss after arteriovenous and arterial clamping for renal warm ischemia in a swine model.

PURPOSE:: To evaluate the glomerular loss after arteriovenous or arterial warm ischemia in a swine model. METHODS:: Twenty four pigs were divided into Group Sham (submitted to all surgical steps except the renal ischemia), Group AV (submitted to 30 minutes of warm ischemia by arteriovenous clamping of left kidney vessels), and Group A (submitted to 30 minutes of ischemia by arterial clamping). Right kidneys were used as controls. Weigh, volume, cortical volume, glomerular volumetric density (Vv[Glom]), volume-weighted glomerular volume (VWGV), and the total number of glomeruli were measured for each organ. RESULTS:: Group AV showed a 24.5% reduction in its left kidney Vv[Glom] and a 25.4% reduction in the VWGV, when compared to the right kidney. Reductions were also observed when compared to kidneys of sham group. There was a reduction of 19.2% in the total number of glomeruli in AV kidneys. No difference was observed in any parameters analyzed on the left kidneys from group A. CONCLUSIONS:: Renal warm ischemia of 30 minutes by arterial clamping did not caused significant glomerular damage, but arteriovenous clamping caused significant glomerular loss in a swine model. Clamping only the renal artery should be considered to minimize renal injury after partial nephrectomies.

Glomerular loss after arteriovenous and arterial clamping for renal warm ischemia in a swine model

ABSTRACT PURPOSE: To evaluate the glomerular loss after arteriovenous or arterial warm ischemia in a swine model. METHODS: Twenty four pigs were divided into Group Sham (submitted to all surgical steps except the renal ischemia), Group AV (submitted to 30 minutes of warm ischemia by arteriovenous clamping of left kidney vessels), and Group A (submitted to 30 minutes of ischemia by arterial clamping). Right kidneys were used as controls. Weigh, volume, cortical volume, glomerular volumetric density (Vv[Glom]), volume-weighted glomerular volume (VWGV), and the total number of glomeruli were measured for each organ. RESULTS: Group AV showed a 24.5% reduction in its left kidney Vv[Glom] and a 25.4% reduction in the VWGV, when compared to the right kidney. Reductions were also observed when compared to kidneys of sham group. There was a reduction of 19.2% in the total number of glomeruli in AV kidneys. No difference was observed in any parameters analyzed on the left kidneys from group A. CONCLUSIONS: Renal warm ischemia of 30 minutes by arterial clamping did not caused significant glomerular damage, but arteriovenous clamping caused significant glomerular loss in a swine model. Clamping only the renal artery should be considered to minimize renal injury after partial nephrectomies.

Estudo morfométrico de rins em felinos domésticos (Felis catus) / Morphometric study on the kidneys of domestic cats (Felis catus)

A saúde renal tem ao longo dos últimos anos chamado atenção dos médicos veterinários, pois o comprometimento deste órgão na insuficiência renal crônica se apresenta como a maior causa de morbidade e mortalidade em felinos. O presente estudo propõe a caracterização biométrica dos rins de gatos, Felis catus, sob os aspectos macroscópicos (comprimento, altura, largura, peso e volume), mesoscópico (altura do córtex e da medula, e a sua inter-relação) e microscópica (volume glomerular), a fim de se estabelecer possíveis diferenças decorrentes de idade, sexo e simetria bilateral. Foram utilizados, rins de 30 animais da espécie Felis catus (gato deméstico), sendo 15 machos e 15 fêmeas com idade variando entre 3 meses a 15 anos, divididos em três grupos: grupo 1 (3-9 meses), grupo 2 (3-5 anos) e grupo 3 (acima de 10 anos)...

Renal health has over the last few years called attention of veterinarians, because the chronic failure of the kidneyu is a major cause of morbidity and mortality in cats. This paper proposes biometric characterization of the kidneys from cats, Felis catus, on the macroscopic aspects (length, height, width, weight and volume), mesoscopic (height of the cortex and medulla, and their inter-relationship) and microscopic (glomerular volume) in order to establish possible differences arising from age, gender and bilateral symmetry. It where used 30 kidneys of animals from specie Felis catus (cat), 15 males and 15 females with ages between 3 months to 15 years, divided into three groups: group 1 (3-9 months), group 2 (3 - 5 years) and group 3 (above 10 years)...

Assessment of glomerular and tubular function.

At birth, GFR and tubular function of neonates is compromised as compared to older children and adults. These functions are even less developed in premature infants. These facts have a direct bearing on drug dosing, fluid and electrolyte administration, and maintenance of acid-base balance in neonates. Although many detailed methods of assessing renal functions have been provided in this article, laboratory and radiologic studies available in most healthcare facilities are often sufficient to provide a clinically relevant data in most patients, including neonates.

Comparative study of the kidneys from dystrophic mice

The Duchenne Muscular Dystrophy (DMD) is a recessive genetic disease linked to chromosome X. Thisdisease is characterized by an absence or dysfunction in the expression of dystrophin. Experimental modelsmdxare widely used for the development of research addressing the DMD. The objective of this research is tocontribute to a detailed study of possible renal morphological changes resulting from DMD. We used five pairsof kidneys frommdxmice and five from normal mice, which were subjected to measurement, light microscopy,and scanning electron microscopy. The morphological findings of kidneys frommdxmice are within thepatterns described in animal studies with severe dehydration, which exhibit signs of diffuse hemorrhage inthe cortical and medullary area, while the glomeruli in the cortical region showed a decrease in urinary space,located between the Bowman’s capsule and the inner cell mass of the glomeruli. However, future experimentswith animals in different ages can assist in the proving of the morphological changes found here.

Aging in the glomerulus.

Kidney function declines with age in the majority of the population. Although very few older people progress to end stage, the consequences of doing so are burdensome for the patient and very expensive for the society. Although some of the observed decline is likely due to changes in the vasculature, much is associated with the development of age-associated glomerulosclerosis. This article will review the well-established structural and functional changes in the glomerulus with age. The role of calorie restriction in modifying age-related pathology will be discussed. The importance of the podocyte as a critical cell in the aging process is considered using animal models and human biopsy material. Newer data on changes in gene expression driven by nuclear factor kappa beta (NFkB) and possible changes in biology in the glomerulus are discussed. The relationship between pathways involved in aging and the decline in kidney function is reviewed. There is speculation on the significance of these changes in relation to normal and pathological aging.

Magnetic resonance histology of age-related nephropathy in the Sprague Dawley rat.

Magnetic resonance histology (MRH) has become a valuable tool in evaluating drug-induced toxicity in preclinical models. However, its application in renal injury has been limited. This study tested the hypothesis that MRH could detect image-based biomarkers of chronic disease, inflammation, or age-related degeneration in the kidney, laying the foundation for more extensive use in evaluating drug toxicity. We examined the entire intact kidney in a spontaneous model of chronic progressive nephropathy. Kidneys from male Sprague Dawley rats were imaged at 8 weeks (n = 4) and 52 weeks (n =4) on a 9.4 T system dedicated to MR microscopy. Several potential contrast mechanisms were explored to optimize the scanning protocols. Full coverage of the entire kidney was achieved with isotropic spatial resolution at 31 microns (voxel volume = 30 pL) using a gradient recalled echo sequence. Isotropic spatial resolution of 15 microns (voxel volume < 4 pL) was achieved in a biopsy core specimen. Qualitative age-related structural changes, such as renal cortical microvasculature, tubular dilation, interstitial fibrosis, and glomerular architecture, were apparent. The nondestructive 3D images allowed measurement of quantitative differences of kidney volume, pelvis volume, main vessel volume, glomerular size, as well as thickness of the cortex, outer medulla, and inner medulla.

[Renal physiology]. / Physiologie rénale.

The kidneys are responsible for the urinary excretion of uremic toxins and the regulation of several body systems such as intra and extracellular volume status, acid-base status, calcium and phosphate metabolism or erythropoiesis. They adapt quantitative and qualitative composition of the urine to keep these systems in balance. The flow of plasma is filtered in the range of 120 mL/min, and depends on the systemic and renal hemodynamics which is subject to self-regulation. The original urine will then be modified in successive segments of the nephron. The proximal nephron is to lead the massive reabsorption of water and essential elements such as sodium, bicarbonates, amino-acids and glucose. The distal nephron includes the distal convoluted tubule, the connector tube and the collecting duct. Its role is to adapt the quality composition of urine to the needs of the body.

Advantages and controversies in the era of intrarenal volumetry.

BACKGROUND/AIMS: Ultrasound is the preferred imaging modality in nephrology. In many kidney diseases, however, more accurate methods are needed to distinguish between relevant intrarenal structures. MRI could be a solution, although the use of MRI contrast has caused severe complications in some cases, and invasive kidney biopsies may follow, even though such small specimens traditionally provide inaccurate quantitative data. We evaluated the usefulness of MRI volumetry and quantitative kidney biopsies to assess glomerular number and volume as well as cortical volume. METHODS: We specifically highlight an experimental study in which different MRI scans were performed in healthy pigs as well as pigs with unilateral ureteral obstruction to assess intrarenal volume. Single-kidney glomerular filtration rate (GFR) was measured using ureteral cannulation and (51)Cr-EDTA. Kidney biopsies were taken and evaluated employing stereological techniques to measure number and volume of glomeruli. Pigs were sacrificed and kidneys were removed for stereological analysis. Non-contrast-based MRI intrarenal volumes were - without significant difference to intrarenal volumes - obtained using contrast-enhanced MRI and ex vivo techniques. RESULTS: Kidney biopsies gave valid estimates regarding quantitative parameters, such as mean number and volume of glomeruli in the cortex. Different structural parameters correlated with kidney GFR with high, although varying, correlation coefficients. CONCLUSION: Non-contrast MRI is suitable for estimating intrarenal volumes in healthy and diseased kidneys. We advocate further research in diagnostic modalities combining MRI and biopsies. Major challenges are the cortical architecture and heterogeneous distribution of glomeruli within the kidney.

Requirement for class II phosphoinositide 3-kinase C2alpha in maintenance of glomerular structure and function.

An early lesion in many kidney diseases is damage to podocytes, which are critical components of the glomerular filtration barrier. A number of proteins are essential for podocyte filtration function, but the signaling events contributing to development of nephrotic syndrome are not well defined. Here we show that class II phosphoinositide 3-kinase C2α (PI3KC2α) is expressed in podocytes and plays a critical role in maintaining normal renal homeostasis. PI3KC2α-deficient mice developed chronic renal failure and exhibited a range of kidney lesions, including glomerular crescent formation and renal tubule defects in early disease, which progressed to diffuse mesangial sclerosis, with reduced podocytes, widespread effacement of foot processes, and modest proteinuria. These findings were associated with altered expression of nephrin, synaptopodin, WT-1, and desmin, indicating that PI3KC2α deficiency specifically impacts podocyte morphology and function. Deposition of glomerular IgA was observed in knockout mice; importantly, however, the development of severe glomerulonephropathy preceded IgA production, indicating that nephropathy was not directly IgA mediated. PI3KC2α deficiency did not affect immune responses, and bone marrow transplantation studies also indicated that the glomerulonephropathy was not the direct consequence of an immune-mediated disease. Thus, PI3KC2α is critical for maintenance of normal glomerular structure and function by supporting normal podocyte function.

Having one kidney does not accelerate the rate of development of diabetic nephropathy lesions in type 1 diabetic patients.

OBJECTIVE: Reduced nephron number is hypothesized to be a risk factor for chronic kidney disease and hypertension. Whether reduced nephron number accelerates the early stages of diabetic nephropathy is unknown. This study investigated whether the rate of development of diabetic nephropathy lesions was different in type 1 diabetic patients with a single (transplanted) kidney compared with patients with two (native) kidneys. RESEARCH DESIGN AND METHODS: Three groups of volunteers were studied: 28 type 1 diabetic kidney transplant recipients with 8-20 years of good graft function, 39 two-kidney patients with duration of type 1 diabetes matched to the time since transplant in the one-kidney group, and 30 age-matched normal control subjects. Electron microscopic morphometry was used to estimate glomerular structural parameters on 3.0 +/- 1.4 glomeruli per biopsy. RESULTS: In the one- versus two-kidney diabetic subject groups, respectively, serum creatinine (means +/- SD 1.3 +/- 0.4 vs. 0.9 +/- 0.2 mg/dl; P < 0.001), systolic blood pressure (133 +/- 13 vs. 122 +/- 11 mmHg; P < 0.001), and albumin excretion rate (median [range] 32.1 microg/min [2-622] vs. 6.8 microg/min [2-1,495]; P = 0.006) were higher. There were no differences in the one- versus two-kidney diabetic subject groups, respectively, in glomerular basement membrane width (median [range] 511 nm [308-745] vs. 473 nm [331-814]), mesangial fractional volume (mean +/- SD 0.30 +/- 0.06 vs. 0.27 +/- 0.07), mesangial matrix fractional volume (0.16 +/- 0.05 vs. 0.16 +/- 0.06), and mesangial matrix fractional volume per total mesangium (0.61 +/- 0.07 vs. 0.64 +/- 0.09). However, these glomerular structural parameters were statistically significantly higher in both diabetic subject groups compared with normal control subjects. Results were similar when patients receiving ACE inhibitors were excluded from the analyses. CONCLUSIONS: Reduced nephron number is not associated with accelerated development of diabetic glomerulopathy lesions in type 1 diabetic patients.

Tetraspan proteins: regulators of renal structure and function.

PURPOSE OF REVIEW: Members of the tetraspan family are widely expressed and poorly understood. An emerging literature suggests that through their interactions with other membrane proteins they play central or regulatory roles in a wide variety of physiological processes. This review will discuss selected tetraspan complexes and highlight their relevance to epithelial cells and the kidney. RECENT FINDINGS: Tetraspans regulate the signaling and trafficking properties of their partner proteins. Tetraspan complexes with integrin molecules, for example, modulate cell adhesion and mobility. Perturbations of tetraspan-integrin assemblies can have dramatic impacts on renal tissue morphogenesis, resulting in a disruption of normal glomerular architecture and selectivity. Tetraspan interactions with renal ion transport proteins appear to affect transporter function by enhancing or inhibiting the endocytic internalization of their transport protein partners. SUMMARY: Tetraspans constitute a novel class of proteins whose capacity to alter the cell biological and functional properties of their membrane protein partners is likely to have wide ranging and important physiological ramifications.

Applicability of the glomerular size distribution coefficient in assessing human glomerular volume: the Weibel and Gomez method revisited.

Changes in glomerular volume (V(glom)) play an important role in the initiation and progression of various glomerulopathies. Estimation of V(glom) in the normal kidney provides baseline values for studies of glomerular hypertrophy in disease. The traditional model-based method of Weibel and Gomez is widely applied to estimate V(glom) in clinical biopsy specimens. Assumptions of glomerular size distribution and shape required by this method are potential sources of bias that have not been verified. We evaluated the applicability of the glomerular size distribution coefficient in estimating V(glom) in human kidneys. V(glom) of 720 non-sclerotic glomeruli in histologically normal kidneys of 24 males (20-69 years) was estimated by the unbiased disector/Cavalieri approach. Accurate glomerular diameters were calculated from Cavalieri estimates of V(glom) assuming glomerular sphericity. The coefficients of variation (CV) of glomerular diameters were compared with the corresponding values of the size distribution coefficient predicted by the Weibel and Gomez method. Mean (SD) glomerular diameter was 201 (28) mm (range 110-276 mm). The CV of glomerular diameter within each kidney ranged from 4.9 to 14.6%. Corresponding glomerular size distribution coefficients predicted by the formula of Weibel and Gomez ranged from 1.00 to just 1.03. The value of the size distribution coefficient required by the Weibel and Gomez technique when estimating V(glom) in normal human kidneys is remarkably constant. This is despite large variations in V(glom). Future studies should examine the extent of bias introduced by the glomerular shape assumptions of this method.

An in vivo rat model simulating imaging of human kidney by diagnostic ultrasound with gas-body contrast agent.

One kidney of anesthetized rats was imaged by diagnostic ultrasound with contrast agent under conditions simulating both the geometry and the attenuation encountered during human perfusion imaging. Contrary to earlier predictions, glomerular capillary rupture with blood loss into Bowman's space and proximal tubules occurred in our clinically relevant model system. Quantitative analysis of histologic sections showed that 37 +/- 5% of the glomeruli at the center of the scan plane had blood cells in Bowman's space after imaging for 1 min with 1.8 MPa (mechanical index equivalent, MIe = 1.5) with a 1 s image trigger interval during IV injection of 10 microl/kg/min of Definity contrast agent (as recommended by the manufacturer). This percentage decreased rapidly with decreasing peak rarefactional pressure amplitude to an apparent threshold of 0.73 MPa (MIe = 0.6). The percentage of glomeruli with hemorrhage decreased in proportion to dose when reduced below the recommended value, but leveled-off at doses above it. The percentage of glomerular hemorrhage increased with increasing numbers of image exposures, with an initial rate of 1.1% per image. The glomerular hemorrhage also depended on the frame trigger interval with no hemorrhage evident for continuous imaging but a maximal effect for trigger intervals greater than about 1 s. These results indicated that there is a potential for clinical diagnostic ultrasound with contrast agent to induce glomerular hemorrhage.

Para-capillary electron-dense deposits reduce glomerular filtration in patients with primary glomerular diseases.

BACKGROUND: Electron-dense deposits are often found around glomerular capillary lumens in patients with glomerulonephritis, forming a portion of the blood-urine barrier (BUB). METHODS: Four hundred and four patients with primary glomerular diseases or donors for living-related kidney transplantation who underwent both percutaneous renal biopsy and renal clearance tests were included in the study. Sodium thiosulfate and paraamino hippurate double-clearance studies were performed with catheterized urinary collection. The filtration fraction (FF) was determined as follows: FF = sodium thiosulfate clearance/paraamino hippurate clearance (CPAH: ). Histomorphometric analyses were performed in 53 patients with overt para-capillary electron-dense deposits (PCEDD) by electron microscopic observations. RESULTS: Patients with membranous nephropathy and membranoproliferative glomerulonephritis showed significantly lower levels of FF than the donors for living-rebated kidney transplantation (normal controls). FF levels were significantly lower in patients with PCEDD than in those without (P < 0.001), while the levels of mean blood pressure and CPAH: were comparable in the two groups. The PCEDD/BUB ratio demonstrated a significant negative correlation with FF (P < 0.0001; r(2) = 0.331). Patients with a ratio of 0.5 or more showed significantly lower FF levels than those with a ratio of 0.25 or less. CONCLUSIONS: PCEDD significantly affected FF levels in patients with primary glomerular diseases. FF may not be an accurate indicator of intraglomerular blood pressure in patients with overt PCEDD.

Localization of tubular uptake segment of filtered Cystatin C and Aprotinin in the rat kidney.

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.

Compensatory kidney growth in estrogen receptor-alpha null mice.

Females are relatively protected in many progressive kidney diseases. Processes of kidney scarring and growth are intricately linked, and female kidneys are smaller than male kidneys. To better understand links between sex, growth, and the kidney, we examined compensatory kidney growth (CKG) after uninephrectomy (Unx) in wild-type and estrogen receptor-alpha null mice (ERKO). Mice (10 wk old) underwent Unx or sham procedure, with removal of all remaining kidney(s) 48 h later. Studies included kidney weight, renal content of protein, DNA, and insulin-like growth factor-I (IGF-I), serum IGF-I, mean glomerular area, and immunostaining for proliferating cell nuclear antigen (PCNA). Sham Unx produced no differences between left and right kidneys. Unx altered kidney weight, glomerular area, DNA content, IGF-I content, and PCNA regardless of sex or genotype. Females showed greater increases in kidney weight (26 vs. 19%) and glomerular area (73 vs. 51%) than males. Differences in kidney weight were restricted to wild-type females (32% increase); ERKO females showed an increase in kidney weight similar to males (19%). Genotype did not influence glomerular growth in this model. Both male and female mice exhibit hyperplastic growth 48 h after Unx, with more pronounced enlargement in females. Lack of estrogen receptor-alpha is associated with reduced CKG in females, probably via suppression of proliferation. ERKO mice did not demonstrate any alterations in compensatory glomerular enlargement. Kidney IGF-I content doubled after Unx, regardless of sex or genotype, implicating other mechanisms with regard to these findings.

Efeitos renais da haploinsuficiência do gene Pkd1 (Polycystic kidney disease 1) em camundongos / Renal effects of Pkd1 gene haploinsufficiency in mice

Vários estudos mostram que na doença renal policística autossômica dominante os cistos surgem a partir de um mecanismo de "dois-golpes". A patogênese das manifestações não-císticas, contudo, é pouco compreendida. Neste estudo usamos uma linhagem de camundongos endogâmica com uma mutação nula em Pkd1, onde animais heterozigotos apresentam formação cística renal mínima até 40 semanas de idade. O clearance de inulina e o número de glomérulos foram menores em machos Pkd1+/- que Pkd1+/+, enquanto o volume glomerular médio foi maior em heterozigotos. A excreção urinária de NO2/NO3 não diferiu significantemente entre os dois grupos. Avaliamos a osmolalidade urinária máxima em machos e fêmeas Pkd1+/- and Pkd1+/+, porém não foi detectada diferença significante entre os grupos heterozigoto e selvagem. Nossos resultados oferecem evidência direta de que a haploinsuficiência de Pkd1 resulta em anormalidades anatômicas e funcionais renais e sugerem que o estado haploinsuficiente de Pkd1 possa resultar na redução do número de néfrons por diminuir a ramificação tubular renal durante a nefrogênese.

Several studies show that in autosomal dominant polycystic kidney disease cysts arise through a "two-hit" mechanism. The pathogenesis of non-cystic features, however, is poorly understood. In this study we used an inbred mouse line with a null mutation of Pkd1, where heterozygotes had minimal renal cyst formation up to 40 weeks of age. Inulin clearance and the number of glomeruli were lower in Pkd1+/- than in Pkd1+/+ males, while a higher average glomerular volume was observed in heterozygotes. The urinary excretion of NO2/NO3 did not significantly differ between the two groups. Maximal urinary osmolality was evaluated in Pkd1+/- and Pkd1+/+ males and females, but no significant difference was detected between the heterozygous and the wild type groups. Our results provide direct evidence that haploinsufficiency for Pkd1 results in anatomic and functional abnormalities of the kidney and suggest that Pkd1 haploinsufficiency may result in a reduced number of nephrons by diminishing renal tubule branching during nephrogenesis.