The renal medullary endothelin system in control of sodium and water excretion and systemic blood pressure

PURPOSE OF REVIEW: Endothelin-1 is a multifunctional peptide that is produced by the kidney and may regulate a variety of renal functions. This review discusses recent developments in understanding the role of the medullary endothelin-1 system in regulating renal salt and water excretion and systemic blood pressure. RECENT FINDINGS: The renal medulla is the major site of endothelin-1 synthesis and receptor expression in the kidney. Endothelin-1 in vitro can inhibit sodium or water transport in the collecting duct and thick ascending limb through autocrine pathways. Endothelin-1 also can increase medullary blood flow. These effects of endothelin-1 are partially mediated by nitric oxide and cyclooxygenase metabolites which are produced by most medullary cells. Mice with collecting duct-specific knockout of the endothelin-1 gene have impaired sodium excretion in response to sodium loading and have hypertension which worsens with high salt intake. The mice also have heightened sensitivity to vasopressin and decreased ability to excrete an acute water load. Mice with collecting duct-specific endothelin A receptor knockout have normal blood pressure and sodium excretion, but have reduced vasopressin responsiveness. Medullary endothelin-1 content is reduced in many forms of experimental hypertension. SUMMARY: Medullary endothelin-1 regulates renal sodium and water transport and medullary blood flow. In particular, the medullary collecting duct is important in this process, but the medullary endothelin system involves complex interactions, through autocrine and paracrine pathways, between most cell types in the region. Medullary endothelin-1 is fundamentally important in physiologic regulation of renal sodium and water excretion and maintenance of normal systemic blood pressure.     

Uric acid crystal binding to renal inner medullary collecting duct cells in primary culture

Attachment of microcrystals to cellular membranes may be an important component in the pathophysiology of urolithiasis. This study characterizes the concentration-dependent binding of uric acid crystals to rat renal inner medullary collecting duct cells in primary culture. Collecting duct cell cultures grew as monolayers with interspersed aggregates of rounded cells. Cultures were incubated with 14C-uric acid crystals, and the crystals that bound were quantitated by adherent radioactivity. Uric acid crystal adherence demonstrated concentration dependent saturation with a 1/alpha value (maximum micrograms of crystals adhering to 1 cm2 of binding area) of 645 micrograms/cm2. The beta values (fraction of cross-sectional area which bound crystals) of uric acid (mean = 0.15) and calcium oxalate monohydrate (mean = 0.13) crystals did not differ significantly. Uric acid crystal binding was inhibited by pre-bound calcium oxalate monohydrate crystals in a concentration dependent manner. These data suggest that uric acid and calcium oxalate crystals exhibit similar binding patterns to rat renal inner medullary collecting duct cells in primary culture.     

Potential physiologic roles for epidermal growth factor in the kidney

Epidermal growth factor (EGF) is a 53-amino acid polypeptide that is known to produce a number of biologic effects both in vitro and in vivo. High concentrations of EGF are found in urine, and high concentrations of prepro-EGF mRNA have been detected in kidney, localized to thick ascending limb of Henle (TALH) and distal convoluted tubule. Specific high-affinity EGF receptors have been demonstrated in mesangial cells, proximal tubule, and cortical and inner medullary collecting duct, as well as in medullary interstitial cells. In the proximal tubule, EGF binding and EGF receptor-associated tyrosine kinase activity are localized to basolateral membrane, and functional responses in collecting duct are observed only with basolateral administration of EGF. A number of renal responses to administration of EGF have recently been described, including modulation of glomerular hemodynamics, renal metabolism, tubular transport functions, and eicosanoid synthesis. In addition, EGF has been shown to be a potent mitogen in vitro for a variety of cell types in the kidney and may be an important mediator of renal repair following injury.     

Effect of epidermal growth factor on the developing rat renal papilla

BACKGROUND: Apoptosis plays an important role in the morphogenesis of the renal papilla. During kidney development, ATL is derived from the TAL in the inner medulla by apoptotic deletion of a fraction of TAL cells and the transformation of the remaining TAL cells. EGF is an important regulator of apoptosis in the kidney. HYPOTHESIS: Exogenously administered EGF in postnatal rat affects renal papilla growth with cell proliferation and apoptosis in the loop of Henle. METHODS: Rat pups received subcutaneous injections of EGF (0.3 microg/g body weight) or saline four times a day from after birth. Rats were sacrificed and the kidneys were preserved for immunohistochemistry on day 4 and day 7. The TAL was identified with antibody directed against Na-K-ATPase or BSC1, and type A intercalated cells were identified with antibody to anion exchanger 1 (AE1). Apoptosis was detected with TUNEL method, and cell proliferation with immunostaining for PCNA. RESULTS--PRIMARY AND SECONDARY: EGF treatment resulted in the following: (1) reduced kidney weight; (2) shortened length of renal papilla; (3) delayed transformation of the cuboidal epithelium into the squamous epithelium of the ATL; (4) delayed elimination of type A intercalated cells in the medullary collecting duct; (5) decreased both apoptotic index and PCNA-positive cells in the TAL and the collecting duct of the renal medulla. CONCLUSION: These findings suggest that exogenous EGF delays the development of loop of Henle in the renal papilla by reducing both apoptosis and cell proliferation.     

Diabetes and radiocontrast media increase endothelin converting enzyme-1 in the kidney

Plasma endothelin-1 levels rise in diabetes and after exposure to contrast media suggesting a role in progressive diabetic and acute radiocontrast nephropathies. Here we studied individual and combined effects of streptozotocin-induced diabetes and contrast media on renal endothelin converting enzyme-1 levels in the rat. In vivo, medullary (but not cortical) endothelin converting enzyme protein gradually increased 4 to 5-fold following the induction of diabetes or after the administration of contrast media but rose 15-fold when diabetic rats were given contrast media. Changes in mRNA expression paralleled those of the protein. Immunohistochemistry confirmed that increased tubular and endothelial cell endothelin converting enzyme-1 were most pronounced in the medulla. In vitro, endothelin-1 levels increased 3-fold following incubation of endothelial cells with media high in glucose or with contrast and 4-fold with their combination. Endothelin converting enzyme-1 protein and mRNA expression changed in a similar pattern while prepro endothelin-1 mRNA increased with each insult but not in an additive way. Our study shows that diabetes and contrast media up-regulate renal medullary endothelin converting enzyme-1 expression and synthesis.     

Na/K-ATPase in intercalated cells along the rat nephron revealed by antigen retrieval

The Na/K-ATPase plays a fundamental role in the physiology of various mammalian cells. In the kidney, previous immunocytochemical studies have localized this protein to the basolateral membrane in different tubule segments. However, intercalated cells (IC) of the collecting duct (CD) in rat and mouse were unlabeled with anti-Na/K-ATPase antibodies. An antigen retrieval technique has been recently described in which tissue sections are pretreated with sodium dodecyl sulfate before immunostaining. This procedure was used to reexamine the presence of Na/K-ATPase in IC along the rat nephron using monoclonal antibodies against the Na/K-ATPase alpha-subunit. Subtypes of IC along the nephron were identified by their distinctive staining with polyclonal and monoclonal antibodies to the 31-kD vacuolar H+ -ATPase subunit, whereas principal cells (PC) were labeled with a polyclonal antibody to the water channel aquaporin-4 (AQP-4). In PC, the Na/K-ATPase and AQP-4 staining colocalized basolaterally. In contrast to previous reports, we found that IC of all types showed basolateral labeling with the anti-Na/K-ATPase antibody. The staining was quantified by fluorescence image analysis. It was weak to moderate in IC of cortical and outer medullary collecting ducts and most intense in IC of the initial inner medullary collecting duct. IC in the initial inner medulla showed a staining intensity that was equivalent or stronger to that in adjacent principal cells. Models of ion transport at the cellular and epithelial level in rat kidney, therefore, must take into account the potential role of a basolateral Na/K-ATPase in intercalated cell function.     

缺血再灌流肾组织内皮素—1及其受体基因表达的实验研究

为了探究内皮素１（ＥＴ１）对肾功能的影响和作用方式，采用斑点杂交和原位杂交方法对大鼠缺血６０分钟再灌注肾组织ＥＴ１及其受体亚型（ＥＴＡ、ＥＴＢ）的基因表达进行了研究。结果发现：再灌流１小时，ＥＴ１、ＥＴＡ、ＥＴＢｍＲＮＡ均明显升高；再灌流２４小时仍维持较高水平。ＥＴ１和ＥＴＡｍＲＮＡ杂交信号再灌流３小时达高峰。ＥＴ１ｍＲＮＡ主要分布肾皮质小血管内皮细胞、髓质肾小管和集合管，ＥＴＡ受体ｍＲＮＡ则分布于上述小血管的平滑肌细胞。ＥＴＢ受体ｍＲＮＡ于再灌流６小时达高峰，主要分布髓质肾小管、集合管。说明缺血再灌流肾内皮素受体亚型上调在皮质以ＥＴＡ为主，在髓质以ＥＴＢ为主，分别与增强表达的ＥＴ１结合导致肾皮质缺血和水钠代谢异常。     

Experimental tests of three-dimensional model of urinary concentrating mechanism.

Recently, a new model of the urinary concentrating process has been proposed that takes into account the three-dimensional architecture of the renal medulla. Under the assumptions of the model, computer simulations predicted significant axial osmolality gradients in the inner medulla without active transport by the inner medullary loop of Henle. Two of the model assumptions (which constitute hypotheses for this study) were: (1) the osmotic water permeability of the initial part of the inner medullary collecting duct (initial IMCD) is very low even in the presence of vasopressin; and (2) there is significant lateral separation of structures such that thin descending limbs are far from the collecting ducts at the same inner medullary level. The first hypothesis was addressed by perfusing rat initial IMCD segments in vitro and measuring osmotic water permeability. With the osmotic gradient oriented as predicted by the model (lumen greater than bath), vasopressin increased the osmotic water permeability from 286 to 852 microns/s. Three additional series of experiments confirmed the high water permeability in the presence of vasopressin. The second hypothesis was addressed by morphometric analysis of histologic cross-sections of the rat renal medulla. Mean distances of descending limbs to the nearest adjacent collecting duct were very small throughout the inner medulla (less than 6 microns) and substantially less than in the outer medulla (28 microns). It was concluded that the data are inconsistent with both hypotheses and therefore do not support the feasibility of the "three-dimensional" model of the renal inner medulla. The axial distributions of loops of Henle and collecting ducts in the rat renal medulla are also reported.     

Role and identification of protein kinase A anchoring proteins in vasopressin-mediated aquaporin-2 translocation

The antidiuretic hormone arginine vasopressin (AVP) regulates water reabsorption in renal principal cells by inducing a cAMP/protein kinase A-dependent translocation of water channels [aquaporin-2 (AQP2)] from intracellular vesicles into the apical cell membranes. Using primary cultured rat inner medullary collecting duct (IMCD) cells, it has been shown that AQP2 translocation in response to AVP stimulation occurs only if protein kinase A (PKA) is anchored to PKA anchoring proteins (AKAPs), which are present in various subcellular compartments. The identity of the AKAPs involved has not yet been elucidated. One potential candidate is a new splice variant of AKAP18, namely AKAP18 delta.     

Renal concentrating and diluting function in deficiency of specific aquaporin genes

Aquaporins (AQP) are water-transporting proteins expressed in many fluid-transporting epithelia and endothelia. In kidney, AQP1 is expressed in plasma membranes of proximal tubule, thin descending limb of Henle and descending vasa recta, AQP2 in collecting duct luminal membrane, AQP3 and AQP4 in collecting duct basolateral membrane, AQP6 in intercalated cells, and AQP7 in the S3 segment of proximal tubule. Human mutations in AQP2 cause hereditary non-X-linked nephrogenic diabetes insipidus. Transgenic mice lacking the renal aquaporins have been useful in defining their role. Mice deficient in AQP1 are polyuric and unable to form a concentrated urine because of defective proximal tubule fluid absorption and countercurrent multiplication. Mice lacking AQP3 are markedly polyuric due to low water permeability across the cortical and outer medullary collecting duct. However, mice lacking AQP4, which is expressed mainly in inner medullary collecting duct, manifest only a mild defect in maximum urinary concentrating ability. The aquaporin null mice have normal urinary diluting ability. From many renal and extrarenal phenotype studies of aquaporin null mice, we conclude that aquaporins are important for rapid near-isosmolar transepithelial fluid absorption/secretion and for rapid vectorial water movement driven by osmotic gradients. The renal phenotype in aquaporin null mice suggests the utility of aquaporin blockers as novel aquaretic-diuretic agents.     

Expression of the ammonia transporter, rh C glycoprotein, in normal and neoplastic human kidney

Recent studies have identified the presence of a novel Mep/Amt/Rh glycoprotein family of proteins that may play an important role in transmembrane ammonia transport. One of the mammalian members of this family, Rh C glycoprotein (RhCG), transports ammonia, is expressed in distal nephron sites that are critically important for ammonia secretion, exhibits increased expression in response to chronic metabolic acidosis, and originally was cloned as a tumor-related protein. The purpose of our studies was to determine the localization of RhCG in the normal and neoplastic human kidney. Immunoblot analysis of human renal cortical protein lysates demonstrated RhCG protein expression with a molecular weight of approximately 52 kD. Immunohistochemistry revealed both apical and basolateral Rhcg expression in the distal convoluted tubule, connecting segment, and initial collecting tubule and throughout the collecting duct. Co-localization with calbindin-D28k, H(+)-ATPase, aquaporin-2, and pendrin showed that distal convoluted tubule and connecting segment cells, A-type intercalated cells, and non-A, non-B cells express RhCG and that B-type intercalated cells, principal cells, and inner medullary collecting duct cells do not. In renal neoplasms, RhCG was expressed by chromophobe renal cell carcinoma and renal oncocytoma but not by clear cell renal cell carcinoma or by papillary renal cell carcinomas. These studies suggest that RhCG contributes to both apical and basolateral membrane ammonia transport in the human kidney. Furthermore, renal chromophobe renal cell carcinoma and renal oncocytoma seem to originate from the A-type intercalated cell.     

The Rh gene family and renal ammonium transport

PURPOSE OF REVIEW: Renal acid-base homeostasis, to a very large extent, depends on renal ammonia production and transport. A putative ammonia transporter family of proteins has recently been identified, and at least two members of this family are expressed in the renal connecting segment and collecting duct. The purpose of this review is to discuss key features of renal ammonia metabolism and transport, with particular emphasis on the transporters involved in this process. RECENT FINDINGS: The putative ammonia transporter family members, RhBG and RhCG, are expressed in the renal connecting segment and collecting duct. Basolateral RhBG is expressed by all cells in the connecting segment and cortical collecting duct, and by intercalated cells in the outer medullary and inner medullary collecting duct. Apical RhCG is expressed in the same distribution and also in the outer stripe of the outer medullary collecting duct principal cells. In all regions, the expression of RhBG and RhCG is greater in intercalated cells than in principal cells. The related protein, RhAG, appears to be an erythroid-specific protein that mediates ammonium/hydrogen ion (NH4/H) exchange. RhBG and RhCG appear to be sodium and potassium ion-independent ammonia transporters. Whether they mediate electrogenic ammonia transport or electroneutral ammonia/hydrogen ion exchange remains an active area of investigation. Finally, transport studies have identified that electroneutral ammonium/hydrogen ion exchange is present in the collecting duct. SUMMARY: The Rh glycoproteins, RhBG and RhCG, appear to mediate important roles in renal ammonia transport, and therefore in acid-base homeostasis.     

Oxalate toxicity in cultured mouse inner medullary collecting duct cells

PURPOSE: Oxalate, a metabolic end product and a major constituent of the majority of renal stones, has been shown to be toxic to renal epithelial cells of cortical origin. However, to our knowledge it is unknown whether inner medullary collecting duct (IMCD) cells, which are physiologically exposed to higher concentrations of oxalate, also behave in a similar manner. MATERIALS AND METHODS: A line of IMCD cells was exposed to oxalate (0.2 to 10 mM) for various time points. Trypan blue, and hematoxylin and eosin stains were used to assess cell morphology and membrane integrity. The production of reactive oxidative species was determined using the nitro blue tetrazolium reaction and crystal violet staining was used to measure cell density. RESULTS: Exposure of IMCD cells to oxalate produced time and concentration dependent changes in the light microscopic appearance of the cells. Long-term exposure to oxalate resulted in alterations in cell viability with net cell loss following exposure to concentrations of 2 mM and greater. Free radical production was time and concentration dependent. Crystal formation occurred in less than 1 hour and cells in proximity to crystals lost membrane integrity. Compared to IMCD cells LLC-PK1 and HK2 cells showed significant toxicity starting at lower oxalate concentrations (0.4 mM and above). CONCLUSIONS: To our knowledge the results provide the first direct demonstration of toxic effects of oxalate in IMCD cells, a line of renal epithelial cells of the inner medullary collecting duct, and suggest that cells lining the collecting duct are relatively resistant to oxalate toxicity.     

Immunocytochemical localization of vitamin D-dependent calcium-binding protein in renal tubules of rabbit, rat, and chick

Vitamin D-dependent calcium-binding protein (CaBP) was localized in tissue sections of kidneys from rabbits, rats, and chicks using antiserum specific for chick intestinal CaBP. In rabbit kidney, CaBP was present in all cells of the distal convoluted tubule and most cells of the connecting tubule. Fewer, but still a majority, of the cells of cortical collecting ducts contained CaBP. The intensity of immunochemical staining and the number of stained cells decreased markedly in medullary collecting ducts, and only a few collecting duct cells contained CaBP at the junction of the inner and outer medulla. In the rat kidney, CaBP was present in all distal convoluted tubule cells, but the immunochemical staining was less intense than in the rabbit. The protein also was found in most connecting tubule cells of the rat; however, only a few collecting duct cells in the superficial corte of the rat contained CaBP. CaBP was essentially absent from mid- to deep-cortical collecting duct cells, while a very few collecting duct cells always contained CaBP at the junction of the inner and outer stripes of the outer medulla. In the chick, CaBP was present in distal convoluted tubule cells as the distal convoluted tubule coursed adjacent to the central vein. CaBP was absent from chick collecting duct cells. In all three species CaBP was not detected in the other portions of the nephron.     

烧伤后肾组织内皮素—1及其受体基因的表达

OBJECTIVE AND METHODS: We studied expression of messenger RNAS for endothelin-1 (ET-1) and its receptor subunits (ETA, ETB) by northern blot hybridization and in situ hybridization in rat kidney following 30% TBSA full thickness burns. RESULTS: Expressions of that ET-1, ETA, and ETB mRNA expression are all evidently increased at 1 hour postburn. ET-1 and ETA reached its maximal mRNA expression at 3 hour postburn. ET-1 mRNA distributed primarily on glomerular endothelial cells and tubular epithelial cells in cortex. ETA mRNA predominantly presented on smooth muscle cells of small arterioles in renal cortex. However, the peak time of ETB mRNA expression is at 6 hour postburn and mainly localized on renal medullary tubular epithelial and collecting duct cells. CONCLUSION: These findings suggest that 1. ET-1 acts principally as a local paracrine/autocrine peptide. 2. Increased ET-1 that activated ETA receptors accentuated expression on vascular smooth muscle cells will result in renal cortex ischemia. 3. Accentuated expression of ETB receptor on tubular epithelial cells may induce renal natriuretic action and reduction of water execretion. 4. Increased ET-1 and its receptors in kidney play an important role in the pathogenesis and development of renal function impairment following severe burn.     

Origin and fate of pendrin-positive intercalated cells in developing mouse kidney

Pendrin is an apical anion exchanger found in type B and nonA-nonB intercalated cells that is involved in bicarbonate secretion. The purpose of this study was to establish the origin and fate of pendrin-positive intercalated cells in the mouse kidney. Using immunohistochemistry, we found that pendrin-positive cells first appeared in the connecting tubule at embryonic day 14 (E14) and subsequently in the medullary collecting duct at E18. Most of the pendrin-positive cells in the connecting tubule were nonA-nonB intercalated cells, wheras those in the medullary collecting duct were type B intercalated cells. In the cortical collecting duct, pendrin-positive cells appeared in the inner part at day 4 after birth and in the outer part at day 7. Pendrin-positive cells gradually disappeared by apoptosis from the inner part of the medullary collecting duct two weeks after birth. Using 5-bromo-2'deoxy-uridine (BrdU) to follow cell proliferation, we determined that selective proliferation of pendrin-positive intercalated cells does not occur; instead, these cells may arise from undifferentiated precursor cells from separate foci, one in the connecting tubule and one in the collecting duct.     

Carcinoma of the Collecting Ducts of Bellini and Renal Medullary Carcinoma: Clinicopathologic Analysis of 52 Cases of Rare Aggressive Subtypes of Renal Cell Carcinoma With a Focus on Their Interrelationship

Carcinoma of the collecting ducts of Bellini and renal medullary carcinoma are rare aggressive neoplasms of putative distal nephron origin. First described in 1949, case reports and review articles constitute a major source of information on collecting duct carcinoma, whereas Davis and colleagues and the pediatric tumor registry have contributed the seminal works on renal medullary carcinoma. Here we present a detailed study of collecting duct carcinoma (n=39) and renal medullary carcinoma (n=13), characterizing these rare neoplasms and analyzing their interrelationship. Both collecting duct carcinoma and renal medullary carcinoma exhibited significant similarities, such as predilection for the right kidney, tumor mass with an epicenter in the renal medulla, and a mean size of 7 cm. Overall, both tumors exhibited a poorly differentiated adenocarcinoma histology with desmoplastic stromal response (100%), inflammatory infiltrate (100%), frequent perinephric extension (collecting duct carcinoma: 97%; renal medullary carcinoma: 83%), lymphovascular invasion (100%), intraluminal mucin (collecting duct carcinoma: 42%; renal medullary carcinoma: 73%), high nuclear grade (97%), overlapping immunoreactivity for Ulex europaeus agglutinin 1 (collecting duct carcinoma: 75%; renal medullary carcinoma:55%), CK7 (collecting duct carcinoma: 44%; renal medullary carcinoma: 71%), and high-molecular weight cytokeratin (collecting duct carcinoma: 26%; renal medullary carcinoma: 29%), and nonimmunoreactivity for Ksp-cadherin. Histologically, collecting duct carcinoma frequently had tubular, tubulopapillary, or irregular glandular architecture, whereas renal medullary carcinoma commonly demonstrated islands of anastomosing tubules and cords forming irregular microcystic spaces. Multiple metastases to the lymph nodes, lung, bone, and liver were observed in both categories at presentation (collecting duct carcinoma: 17%; renal medullary carcinoma: 36%). Only patients with organ-confined small tumors were disease free beyond the median survival time. Differential clinical features between collecting duct carcinoma and renal medullary carcinoma included proclivity for younger male individuals of African ancestry with hemoglobin abnormalities and a shorter median survival of 17 weeks (vs. 44 wk for collecting duct carcinoma) for renal medullary carcinoma. The markedly overlapping clinical features, histology, immunophenotype, metastasis patterns, and uniformly aggressive outcome in collecting duct and renal medullary carcinomas suggest that renal medullary carcinoma is a distinctive clinicopathologic subtype within the entity of collecting duct carcinoma. The extremely poor prognosis and ongoing clinical trials with specific therapeutic protocols argue for their accurate distinction from other renal cell carcinoma subtypes.     

Expression of RhCG,a new putative NH(3)/NH(4)(+) transporter,along the rat nephron

Two non-erythroid members of the erythrocyte Rhesus (Rh) protein family, RhBG and RhCG, have been recently cloned in the kidney. These proteins share homologies with specific NH(3)/NH(4)(+) transporters (Mep/Amt) in primitive organisms and plants. When expressed in a Mep-deficient yeast, RhCG can function as a bidirectional NH(3)/NH(4)(+) transporter. The aim of this study was to determine the intrarenal and intracellular location of RhCG in rat kidney. RT-PCR on microdissected rat nephron segments demonstrated expression of mRNAs encoding RhCG in distal convoluted tubules, connecting ducts, and cortical and outer medullary collecting ducts but not in proximal tubules and thick ascending limbs of Henle's loop. Immunolocalization studies performed on rat kidney sections with rabbit anti-human RhCG 31 to 48 antibody showed labeling of the apical pole of tubular cells within the cortex, the outer medulla, and the upper portion of the inner medulla. All cells within connecting tubules had identical apical staining. In cortical collecting ducts, a subpopulation of cells that has either apical staining (alpha-intercalated cells) or diffuse staining (beta-intercalated cells) for the beta1 subunit of the H(+)-ATPase, was heavily stained at their apical pole with the RhCG antibody while principal cells identified as H(+)-ATPase negative cells showed a faint apical staining for RhCG that was much less intense than in adjacent intercalated cells. In the outer medulla and the upper portion of the inner medulla, RhCG labeling was restricted to a subpopulation of cells within the collecting duct that apically express the beta1 subunit of the H(+)-ATPase, indicating that RhCG expression in medullary collecting ducts is restricted to intercalated cells. No labeling was seen in glomeruli, proximal tubules, and limbs of Henle's loop. Immunoblotting of apical membrane fractions from rat kidney cortex with the rabbit anti-human RhCG 31 to 48 antibody revealed a doublet band at approximatively 65 kD.     

氨转运蛋白Rh b型糖蛋白在低蛋白饮食大鼠肾脏的表达

目的 研究低蛋白饮食对氨转运蛋白Rh b型糖蛋白（Rhbg ）在大鼠肾脏表达的影响。 方法 利用Western印迹分别检测Rhbg在低蛋白饮食组和正常对照组大鼠肾脏皮质、内髓、外髓的表达。利用单标记免疫组化法、双标记免疫组化法和定量免疫组化法检测Rhbg在两组中肾小管主细胞和暗细胞的表达。 结果 与正常对照组比较,在大鼠肾脏皮质,低蛋白饮食组Rhbg 的蛋白水平显著增高（954778±509288比275701±262374,P < 0.05）;在大鼠肾脏内髓和外髓,低蛋白饮食组Rhbg 的蛋白水平无明显变化;在大鼠肾脏皮质集合管主细胞,低蛋白饮食组免疫组化Rhbg表达像素值（1310±357）显著高于对照组（896±154,P < 0.05）;在大鼠肾脏皮质集合管暗细胞,低蛋白饮食组免疫组化Rhbg表达像素值（1550±497）显著高于对照组（926±251,P < 0.05）;在大鼠肾脏内髓集合管和外髓集合管的主细胞或暗细胞中,两组间免疫组化Rhbg表达无显著差异。 结论 饮食中蛋白的限制可能会增加Rhbg在大鼠肾脏皮质集合管的主细胞和暗细胞的表达。