Renal Vascular Development Research Articles

Origins and molecular mechanisms underlying renal vascular development.

Kidneys play a crucial role in maintaining homeostasis within the body, and this function is intricately linked to the vascular structures within them. For vascular cells within the kidney to mature and perform their functions effectively, it is imperative that there is a meticulous and well-coordinated spatial alignment between the nephrons and the intricate network of blood vessels within the organ. This spatial arrangement ensures efficient filtration of blood and regulation of the electrolyte balance, blood pressure, and fluid levels. Additionally, the kidneys play a vital role in the regulation of acid-base balance and the production of hormones involved in erythropoiesis and blood pressure regulation. Thus, the close interaction between the vascular system and the kidney's structural organization is essential for maintaining overall physiological balance and health. This article focuses on the vascular development of the kidneys, summarizing the current understanding of the origin and formation of the renal vasculature, and the crucial molecules involved. By elucidating the cellular and molecular mechanisms governing renal vascular development, this article aims to promote advancements in renal regenerative medicine and provide potential avenues for therapeutic interventions to address kidney disease.

Sex Differences in renal ischemia-reperfusion injury following deletion of Netrin-1 in the kidney

Netrin-1, an angiogenic guidance cue, is heavily expressed in the kidney. Our lab has previously reported that deletion of Netrin-1 in Foxd1-positive stromal progenitors ( Foxd1 GC/+ Netrin1 fl/fl ) results in abnormal vascular patterning during kidney development, including a reduction in interlobar branching. It remains unknown how these changes in vascular patterning affect kidney function and physiology. In the current study, we hypothesized that changes in developmental patterning of the kidney vasculature would protect from renal ischemia-reperfusion injury in a sex-specific manner due to ischemic preconditioning to the kidney tubular cells. To test this hypothesis, we performed warm, bilateral ischemia-reperfusion by clamping the renal pedicles for 26-minutes in wild-type ( Netrin1 fl/fl or Netrin1 fl/+ )(3M, 3F), heterozygous ( Foxd1 GC/+ Netrin1 fl/+ ) (2M, 4F), and mutant ( Foxd1 GC/+ Netrin1 fl/fl ) (3M, 1F) male (M) and female (F) mice. Following 24-hours of reperfusion, blood and kidneys were collected for analysis of kidney injury markers. We found that plasma creatinine (p 2-wayANOVA =0.021) and blood urea nitrogen (p 2-wayANOVA =0.009) were decreased in female mice compared to male mice regardless of genotype. In both male and female mutant mice, there was a decrease in these markers of injury compared to heterozygous and wild-type control mice, although this did not reach statistical significance. In agreement, immunofluorescent staining of kidney injury marker 1 (KIM-1) indicated less tubular injury in female mice compared to males. These data suggest that female mice are less susceptible to ischemic injury than male mice despite both sexes having similar alterations in vascular patterning following deletion of Netrin-1. Further, our results indicate a protective effect of these developmental patterning changes due to Netrin-1 deletion. We suspect that changes in the vasculature in our mutant mice provide a preconditioning-like environment to mediate this effect. Future studies will investigate the underlying mechanisms which drive this injury phenotype between sexes. Understanding how renal vascular development impacts kidney injury is critical to the development of new therapeutic strategies to prevent the sustained injury following periods of ischemia. NIH R01DK121014 to LO This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Mouse Metanephric Mesenchymal Cell–Derived Angioblasts Undergo Vasculogenesis in Three-Dimensional Culture

Invitro models for the investigation of renal vascular development are limited. We previously showed that isolated metanephric mesenchymal (MM) and ureteric bud (UB) cells grown in three-dimensional (3D) matrices formed organoids that consisted of primitive vascular structures surrounding a polarized epithelium. Here, we examined the potential of two principal effectors of vasculogenesis, vascular endothelial growth factor A (VEGF-A), and platelet-derived growth factor B chain (PDGF-BB), to stimulate MM cell differentiation. The results showed that MM cells possess angioblast characteristics by expressing phenotypic markers for endothelial and mesenchymal cells. UB cells synthesize VEGF-A and PDGF-BB proteins and RNA, whereas the MM cells express the respective cognate receptors, supporting their role in directional induction of vasculogenesis. VEGF-A stimulated proliferation of MM cells in monolayer and in 3D sponges but did not affect MM cell migration, organization, or vasculogenesis. However, PDGF-BB stimulated MM cell proliferation, migration, and vasculogenesis in monolayer and organization of the cells into primitive capillary-like assemblies in 3D sea sponge scaffolds invitro. A role for PDGF-BB in vasculogenesis in the 3D MM/UB co-culture system was validated by direct interference with PDGF-BB or PDGF receptor-β cell interactions to implicate PDGF-BB as a primary effector of MM cell vasculogenesis. Thus, MM cells resemble early renal angioblasts that may provide an ideal platform for the investigation of renal vasculogenesis invitro.

Expression of receptor-type protein tyrosine phosphatase in developing and adult renal vasculature.

Renal vascular development is a coordinated process that requires ordered endothelial cell proliferation, migration, intercellular adhesion, and morphogenesis. In recent decades, studies have defined the pivotal role of endothelial receptor tyrosine kinases (RPTKs) in the development and maintenance of renal vasculature. However, the expression and the role of receptor tyrosine phosphatases (RPTPs) in renal endothelium are poorly understood, though coupled and counterbalancing roles of RPTKs and RPTPs are well defined in other systems. In this study, we evaluated the promoter activity and immunolocalization of two endothelial RPTPs, VE-PTP and PTPμ, in developing and adult renal vasculature using the heterozygous LacZ knock-in mice and specific antibodies. In adult kidneys, both VE-PTP and PTPμ were expressed in the endothelium of arterial, glomerular, and medullary vessels, while their expression was highly limited in peritubular capillaries and venous endothelium. VE-PTP and PTPμ promoter activity was also observed in medullary tubular segments in adult kidneys. In embryonic (E12.5, E13.5, E15.5, E17.5) and postnatal (P0, P3, P7) kidneys, these RPTPs were expressed in ingrowing renal arteries, developing glomerular microvasculature (as early as the S-shaped stage), and medullary vessels. Their expression became more evident as the vasculatures matured. Peritubular capillary expression of VE-PTP was also noted in embryonic and postnatal kidneys. Compared to VE-PTP, PTPμ immunoreactivity was relatively limited in embryonic and neonatal renal vasculature and evident immunoreactivity was observed from the P3 stage. These findings indicate 1) VE-PTP and PTPμ are expressed in endothelium of arterial, glomerular, and medullary renal vasculature, 2) their expression increases as renal vascular development proceeds, suggesting that these RPTPs play a role in maturation and maintenance of these vasculatures, and 3) peritubular capillary VE-PTP expression is down-regulated in adult kidneys, suggesting a role of VE-PTP in the development of peritubular capillaries.

Kidney Dysfunction in Adult Offspring Exposed In Utero to Type 1 Diabetes Is Associated with Alterations in Genome-Wide DNA Methylation.

BackgroundFetal exposure to hyperglycemia impacts negatively kidney development and function.ObjectiveOur objective was to determine whether fetal exposure to moderate hyperglycemia is associated with epigenetic alterations in DNA methylation in peripheral blood cells and whether those alterations are related to impaired kidney function in adult offspring.DesignTwenty nine adult, non-diabetic offspring of mothers with type 1 diabetes (T1D) (case group) were matched with 28 offspring of T1D fathers (control group) for the study of their leukocyte genome-wide DNA methylation profile (27,578 CpG sites, Human Methylation 27 BeadChip, Illumina Infinium). In a subset of 19 cases and 18 controls, we assessed renal vascular development by measuring Glomerular Filtration Rate (GFR) and Effective Renal Plasma Flow (ERPF) at baseline and during vasodilatation produced by amino acid infusion.ResultsGlobally, DNA was under-methylated in cases vs. controls. Among the 87 CpG sites differently methylated, 74 sites were less methylated and 13 sites more methylated in cases vs. controls. None of these CpG sites were located on a gene known to be directly involved in kidney development and/or function. However, the gene encoding DNA methyltransferase 1 (DNMT1)—a key enzyme involved in gene expression during early development–was under-methylated in cases. The average methylation of the 74 under-methylated sites differently correlated with GFR in cases and controls.ConclusionAlterations in methylation profile imprinted by the hyperglycemic milieu of T1D mothers during fetal development may impact kidney function in adult offspring. The involved pathways seem to be a nonspecific imprinting process rather than specific to kidney development or function.

Abstract 312: Jagged1 and Morphological Integrity of the Kidney

Renal vascular development is dependent on the participation of renin precursor cells, which localize to areas of new vessel formation and differentiate into smooth muscle cells (SMCs), mesangial and juxtaglomerular (JG) cells. However, the mechanisms that enforce this particular fate of renin precursor cells remain unclear. During embryogenesis, renin cells express a significant number of angiogenic factors including the Notch signaling ligand Jagged1. We therefore hypothesized that Jagged1 is necessary for renin precursors to differentiate into the various mural cells of the renal arterioles and that Jagged1 plays a critical role in renal vascular development. To generate conditional Jagged1 deletion in cells of the renin lineage, we crossed Jagged1 fl/fl mice with Ren1d cre/+ mice. We investigated kidney weight, morphology, and vascular architecture as well as the identity of cells composing the kidney arterioles including renin cells and SMCs. We performed quantitative RT-PCR on kidney cortex mRNA for Jagged1 and Renin expression levels and immunohistochemistry for renin. The expression of Jagged1 mRNA in conditional knockout (cKO) animals was reduced at 2 weeks (68.3% reduction) and 4 weeks (66.5% reduction) compared to control mice. We found no significant differences between control and cKO animals in their kidney weights, renin mRNA expression, and renin staining. The number of renin-positive juxtaglomerular apparatuses (JGA) was 49.25 +/- 3.301 (n=4) and 38.00 +/- 4.830 (n=4) for control and cKO mice respectively under physiologic conditions. Staining for α-smooth muscle actin (α-SMA) demonstrated an overall normal vascular anatomy in cKO kidneys, however there were focal areas containing activated pericytes and injured mesangial cells expressing α-SMA indicating an active fibrotic process. Mason’s trichrome staining confirmed areas of glomerular and interstitial fibrosis in cKO kidneys. This work suggests that while Jagged1 is dispensable for renin expression, its loss in renin cell descendants affects perivascular-interstitial integrity and glomerular structure. The findings indicate an important role of Jagged1 in the maintenance of the morphologic integrity of the kidney.

SDF-1/CXCR4 Signaling Preserves Microvascular Integrity and Renal Function in Chronic Kidney Disease

The progressive decline of renal function in chronic kidney disease (CKD) is characterized by both disruption of the microvascular architecture and the accumulation of fibrotic matrix. One angiogenic pathway recently identified as playing an essential role in renal vascular development is the stromal cell-derived factor-1α (SDF-1)/CXCR4 pathway. Because similar developmental processes may be recapitulated in the disease setting, we hypothesized that the SDF-1/CXCR4 system would regulate microvascular health in CKD. Expression of CXCR4 was observed to be increased in the kidneys of subtotally nephrectomized (SNx) rats and in biopsies from patients with secondary focal segmental glomerulosclerosis (FSGS), a rodent model and human correlate both characterized by aberration of the renal microvessels. A reno-protective role for local SDF-1/CXCR4 signaling was indicated by i) CXCR4-dependent glomerular eNOS activation following acute SDF-1 administration; and ii) acceleration of renal function decline, capillary loss and fibrosis in SNx rats treated with chronic CXCR4 blockade. In contrast to the upregulation of CXCR4, SDF-1 transcript levels were decreased in SNx rat kidneys as well as in renal fibroblasts exposed to the pro-fibrotic cytokine transforming growth factor β (TGF-β), the latter effect being attenuated by histone deacetylase inhibition. Increased renal SDF-1 expression was, however, observed following the treatment of SNx rats with the ACE inhibitor, perindopril. Collectively, these observations indicate that local SDF-1/CXCR4 signaling functions to preserve microvascular integrity and prevent renal fibrosis. Augmentation of this pathway, either purposefully or serendipitously with either novel or existing therapies, may attenuate renal decline in CKD.

SU‐E‐I‐17: Micro CT Analysis of Results of Protein Deficiency On Rat Offspring

Purpose: To evaluate the use of image based metrics in the study of renal vascular development in offspring from low protein fed pregnant rats. Methods: Using a custom, self made micro CT imaging system, we imaged two groups of rat kidneys. The control group consisted of eight one year old Wistar rat offspring exposed to normal maternal rat diet during pregnancy. The second group, of the same breed and age, were eight offspring of a low protein maternal diet during pregnancy. Both groups were euthanized and a warm barium sulfate gelatin suspension was pressure infused into the arterial circulation, creating radio opacity for identification of the arterial vasculature. Approximately one hour after administration, the kidneys were harvested, placed in formalin, then stored at 4°C. Each kidney was imaged using our micro CT machine set from 47 to 55 kVp and 40 to 60 Watts. Results: Clearly the renal vasculature development differed greatly between the two groups. In particular, the protein deficient group developed segmental renal arteries with larger diameters. There was also a minimal difference in the percentage of arteries to tissue volume for the two groups. After examining the 3D reconstructions, we set gray level threshold values for vasculature and for tissue. We then determined the corresponding volumes and volume ratio using LabView. Utilizing the projection images, we also measured the diameter of three vessels near the hilum of the kidney, segmental arteries. On average, the control group and protein deficient artery diameters were 375.71±49.10 516.33±52.58 μm, respectively. A comparison of histograms of the vessel diameters demonstrated a minimally overlapping classification of the two groups of animals. Conclusion: Using our custom made Micro CT scanner images, we were able to quantify the physiological effect of a maternal protein deficient diet on the developmental morphology of the offspring kidney vasculature. Support: NIH Grant 2R01EB002873

Protein‐energy malnutrition during early gestation in sheep blunts fetal renal vascular and nephron development and compromises adult renal function

A nutritionally poor maternal diet can reduce nephron endowment and pre-empt premature expression of markers for chronic renal disease in the offspring. A mechanistic pathway from variation in maternal diet through altered fetal renal development to compromised adult kidney structure and function with adult-onset obesity has not been described. We show that maternal protein-energy malnutrition in sheep blunts nephrogenic potential in the 0.44 gestation (65 days gestation, term ∼147 days) fetus by increasing apoptosis and decreasing angiogenesis in the nephrogenic zone, effects that were more marked in male fetuses. As adults, the low-protein-exposed sheep had reduced glomerular number and microvascular rarefaction in their kidneys compensated for, respectively, by glomerular hypertrophy and increased angiogenic support. In this study, the long-term mild anatomical deficits in the kidney would have remained asymptomatic in the lean state, but when superimposed on the broad metabolic challenge that obesity represents then microalbuminuria and blunted bilateral renal function revealed a long-term physiological compromise, that is only predicted to worsen with age. In conclusion, maternal protein-energy malnutrition specifically impacts fetal kidney vascular development and prevents full functionality of the adult kidney being achieved; these residual deficits are predicted to significantly increase the expected incidence of chronic kidney disease in prenatally undernourished individuals especially when coupled with a Western obesogenic environment.

A model for renal arterial branching based on graph theory

The kidney is one of the most complicated organs in terms of structure and physiology, in part because it is highly vascularized. The renal vascular development occurs through two mechanisms that sometimes overlap: vasculogenesis and angiogenesis. Here, we consider angiogenesis to model the renal arterial tree with the two processes of vascular angiogenesis: sprouting and splitting. We recognize the vessels are not tubes with ends that get glued but physiological factors are relevant into the vascular development. Our contribution integrates the graph theory and physiological information to derive a quantitative model for the vascular tree in the sense that the vertices and edges represent, respectively, a branching point and a vessel. From such a premise, development of the arterial vascular tree of the kidney is mathematically expressed, including physiological processes as the effect of the vascular endothelial growth factor (VEGF) on the vessel length. A definition of the graph is used to visualize the topology of vascular tree in kidney providing physiological information into the edges. Thus, renal arterial branching is modeled as a graph where edges are labeled and oriented.

PECAM-1 isoform-specific regulation of kidney endothelial cell migration and capillary morphogenesis

Platelet endothelial cell adhesion molecule-1 (PECAM-1) has been implicated in angiogenesis through its involvement in endothelial cell-cell and cell-matrix interactions and signal transduction. Recent studies indicate that the cytoplasmic domain of PECAM-1 plays an important role in its cell adhesive and signaling properties. However, the role PECAM-1 isoforms play during angiogenic events such as cell adhesion and migration requires further delineation. To gain insight into the role PECAM-1 plays during vascular development and angiogenesis, we examined the expression pattern of PECAM-1 isoforms during kidney vascularization. We show that multiple isoforms of PECAM-1 are expressed during renal vascular development with different frequencies. The PECAM-1 that lacks exons 14 and 15 (Delta14&15) was the predominant isoform detected in the renal vasculature. To further study PECAM-1 isoform-specific functions we isolated kidney endothelial cells (EC) from wild-type and PECAM-1-deficient (PECAM-1-/-) mice with B(4)-lectin-coated magnetic beads. PECAM-1-/- kidney EC showed reduced migration, inability to undergo capillary morphogenesis in Matrigel, dense peripheral focal adhesions, and peripheral cortical actin distribution compared with wild-type cells. PECAM-1-/- kidney EC secreted increased amounts of fibronectin and decreased amounts of tenascin-C and thrombospondin-1. Reexpression of Delta14&15, but not full-length, PECAM-1 in PECAM-1-/- kidney EC restored cell migration and capillary morphogenesis defects. Thus PECAM-1 may regulate the adhesive and migratory properties of kidney EC in an isoform-specific fashion through modulation of integrin activity and extracellular matrix protein expression. Our results indicate that regulated expression of specific PECAM-1 isoforms may enable EC to accommodate the different stages of angiogenesis.

Kidney development and gene expression in the HIF2α knockout mouse

The hypoxia-inducible transcription factor-2 (HIF2), a heterodimer composed of HIF2alpha and HIF1beta subunits, drives expression of genes essential for vascularization, including vascular endothelial growth factor (VEGF) and VEGF receptor-2 (VEGFR-2, Flk-1). Here, we used a HIF2alpha/LacZ transgenic mouse to define patterns of HIF2alpha transcription during kidney development and maturation. Our results from embryonic heterozygotes showed HIF2alpha/LacZ expression by apparently all renal endothelial cells. At 4 weeks of age, glomerular mesangial and vascular smooth muscle cells were also positive together with endothelial cells. These expression patterns were confirmed by electron microscopy using Bluo-gal as a beta-galactosidase substrate. Small numbers of glomerular and tubular epithelial cells were also positive at all stages examined. Light and electron microscopic examination of kidneys from HIF2alpha null embryos showed no defects in renal vascular development or nephrogenesis. Similarly, the same amounts of Flk-1 protein were seen on Western blots of kidney extracts from homozygous and heterozygous HIF2alpha mutants. To examine responsiveness of HIF2alpha null kidneys to hypoxia, embryonic day 13.5 metanephroi were cultured in room air or in mild (5% O(2)) hypoxia. For both heterozygous and null samples, VEGF mRNA levels doubled when metanephroi were cultured in mild hypoxia. Anterior chamber grafts of embryonic HIF2alpha knockouts were morphologically indistinguishable from heterozygous grafts. Endothelial markers, platelet endothelial cell adhesion molecule and BsLB4, as well as glomerular epithelial markers, GLEPP1 and WT-1, were all expressed appropriately. Finally, we undertook quantitative real-time polymerase chain reaction of kidneys from HIF2alpha null embryos and wild-type siblings and found no compensatory up-regulation of HIF1alpha or -3alpha. Our results show that, although HIF2alpha was widely transcribed by kidney endothelium and vascular smooth muscle, knockouts displayed no detectable deficits in vessel development or VEGF or Flk-1 expression.

Normal renal vascular development is dependant on Crim1 and its regulation of VEGFA.

Crim1 is a cell-surface transmembrane protein expressed in many organ systems in the developing embryo. We have shown that Crim1 binds intracellularly to a number of growth factors including VEGFA and acts to tether the growth factor to the cell surface thereby regulating its release. In the kidney Crim1 is expressed in the arterial vasculature. Our analyses of a Crim1 LacZ knockin mouse line (KST264), which has severe renal defects, have shown that Crim1 is essential for normal glomerular vascular development by regulating the release of VEGFA from the glomerular epithelial cells. We are now investigating the role of Crim1 in the development of the peritubular capillary network. Kidneys from embryonic and adult KST264 homozygotes show defects in the tubules suggesting chronic ischemia. This is supported by studies using hypoxyprobe which has identified ischemia in the tubules of both embryonic and adult homozygotes. In cell-based studies, Crim1 is upregulated along with VEGFA during hypoxia. Immunohistochemistry and electron microscopy of kidney sections from adult and perinatal homozygotes revealed areas in which peritubular capillaries are absent or abnormal. These studies suggest that Crim1 is essential for normal vascular development in the kidney and may act to regulate VEGFA induced angiogenesis. This work was supported by an NHMRC Australia grant (455972).

Neonatal losartan treatment suppresses renal expression of molecules involved in cell-cell and cell-matrix interactions.

Lack of neonatal angiotensin II type 1 receptor (AT(1)) stimulation produces renal abnormalities characterized by papillary atrophy and impaired urinary concentrating ability, but the mechanisms involved are still unclear. DNA microarray was used to identify genes that are differentially expressed in renal medulla in response to neonatal treatment with AT(1) receptor antagonist losartan (30 mg/kg per d), which commenced within 24 h after birth. The data showed that losartan treatment for 48 h downregulated 68 genes, approximately 30% of which encode various components of cytoskeleton and cytoskeleton-associated proteins, extracellular matrix, and enzymes involved in extracellular matrix maturation or turnover. With the use of immunohistochemistry and Western immunoblot, the microarray data were confirmed and it was demonstrated that losartan suppressed renal expression of syndecan 2, alpha-smooth muscle actin, MHC class II, and leukocyte type 12-lipoxygenase by day 4. In addition, losartan inhibited medullary expression of integrin alpha6 and caused relocalization of integrins alpha6 and alpha3. Moreover, losartan inhibited cell proliferation in medullary tubules by day 9, as detected by Ki-67 immunostaining. This study provides new data supporting the contention that a lack of AT(1) receptor stimulation results in abnormal matrix assembly, disturbed cell-cell and cell-matrix interactions, and subsequent abnormal tubular maturation. Moreover, regulation of the expression of leukocyte type 12-lipoxygenase and alpha-smooth muscle actin by the renin-angiotensin system in the immature kidney adds new knowledge toward the understanding of renal vascular development.

Angiopoietin growth factors and Tie receptor tyrosine kinases in renal vascular development.

Angiopoietin-1 (Ang-1) is a secreted growth factor which binds to and activates the Tie-2 receptor tyrosine kinase. The factor enhances endothelial cell survival and capillary morphogenesis, and also limits capillary permeability. Ang-2 binds the same receptor but fails to activate it: hence, it is a natural inhibitor of Ang-1. Ang-2 destabilises capillary integrity, facilitating sprouting when ambient vascular endothelial growth factor (VEGF) levels are high, but causing vessel regression when VEGF levels are low. Tie-1 is a Tie-2 homologue but its ligands are unknown. Angiopoietin and Tie genes are expressed in the mammalian metanephros, the precursor of the adult kidney, where they may play a role in endothelial precursor growth. Tie-1-expressing cells can be detected in the metanephros when it first forms and, based on transplantation experiments, these precursors contribute to the generation of glomerular capillaries. During glomerular maturation, podocyte-derived Ang-1 and mesangial-cell-derived Ang-2 may affect growth of nascent capillaries. After birth, vasa rectae acquire their mature configuration and Ang-2 expressed by descending limbs of loops of Henle would be well placed to affect the growth of this medullary microcirculation. Finally, preliminary data implicate angiopoietins in deregulated vessel growth in Wilms' kidney tumours and in vascular remodelling after nephrotoxicity.

Influence of fetal environment on kidney development.

Several lines of evidence, mostly derived from animal studies, indicate that changes in fetal environment may affect renal development. Besides maternal hyperglycemia or drug exposure, that were recently found to alter nephrogenesis, changes in vitamin A supply to the fetus may prove to be responsible for most of the variations in nephron number found in the population. A low vitamin A status in the fetus may be a major cause of inborn nephron deficit, either as a feature of intrauterine growth retardation or independently of growth retardation. The possibility that vitamin A status may also influence renal vascular development is raised. We suggest that low vitamin A supply to the fetus plays a role in the intrauterine programming of chronic renal disease and hypertension.

Role of angiotensin in renal vascular development

All components of the renin-angiotensin system (RAS) are expressed in the developing kidney in a temporospatial pattern that suggests a role for this system in kidney morphogenesis. Pharmacological blockade of angiotensin actions in fetal and newborn animals results in striking alterations in kidney architecture, including immature glomeruli and papillae, dilated tubuli, and arrested vascular development. Inactivation of angiotensinogen or angiotensin converting enzyme genes in mice results in similar anomalies that begin as subtle alterations in early life and become more pronounced as extrauterine life progresses. However, inactivation of each angiotensin receptor subtype does not result in obvious morphological abnormalities, suggesting functional redundancy at the receptor level. Crossing of mice lacking the various receptor subtypes should be revealing. Overall, the available information suggests that the RAS is necessary for the normal morphological and functional development of the kidney and the preservation of kidney architecture in adult life.

TGF-β type II receptor in rat renal vascular development: Localization to juxtaglomerular cells

To further define the role of transforming growth factor-beta (TGF-beta) receptors in renal vascular development, detailed immunohistochemical studies of TGF-beta receptor expression were performed from gestational day 15 through adulthood. On gestational day 15, TGF-beta type II receptor immunoreactivity was restricted to perirenal stromal and vascular cells. On gestational day 17 TGF-beta type II receptor immunoreactive stromal cells were observed within the kidney, with the same distribution as stromal alpha-smooth muscle actin and renin immunoreactive cells, and intense stromal TGF-beta type II receptor immunoreactivity continued through postnatal day 5. As vascular development progressed, TGF-beta type II receptor, alpha-smooth muscle actin and renin immunoreactivity became progressively restricted to small renal arteries and arterioles. Expression of TGF-beta type II receptors and renin was very intense in afferent glomerular arterioles during postnatal days 5 to 15, and then became progressively restricted only to juxtaglomerular cells in the mature kidney. TGF-beta type I receptor (ALK-5, ALK-1 and ALK-2) immunoreactivity was not detected in stromal or vascular elements during development or in the mature kidney. Intense TGF-beta type II receptor expression in renal stromal vascular smooth muscle cell precursors and developing blood vessels suggests a role for the TGF-beta type II receptors in the formation of the renal vascular smooth muscle compartment. The continued intense expression in juxtaglomerular cells argues for a role in renin synthesis and/or release. The absence of ALK-5, ALK-1, and ALK-2 in developing vascular smooth muscle and mature juxtaglomerular cells indicates that the canonical view of TGF-beta signaling may not hold in these locations.

Renin-expressing cells are associated with branching of the developing kidney vasculature.

To define the relationship between renal vascular development and renin distribution during kidney ontogeny, the complete renal arterial tree of Sprague Dawley rats during fetal (20 d) and postnatal (1 to 90 d) life was microdissected and immunostained for renin. A shift in renin distribution from interlobar and arcuate arteries in the fetus to the afferent arterioles in the adult was observed. In addition, seven types of renin distribution along the afferent arterioles were identified. In type I, renin was distributed continuously along the whole length of the afferent vessel. This pattern was most frequently observed in the fetus. In type II, renin extended upstream from the glomerulus but did not occupy the whole length of the arteriole. This type was relatively constant throughout postnatal life. In type III, renin was present as bands along the afferent vessel; it was most frequently observed in the fetal and early perinatal periods. In type IV, renin was restricted to the "classical" juxtaglomerular localization. It was the most frequent type observed in the adult rat. In type V, no renin was found in the arteriole. It was the second most frequent type observed in the adult rat. In addition, two "mixed" patterns, type III/IV and type III/II, were occasionally observed. The distribution of renin-expressing cells was spatially and temporally associated with the development of blood vessels. Development of a new arterial branch was preceded by the appearance of renin-expressing cells at the point of branching. This was followed by an outpouching of the arterial wall that progressively elongated to form a new arteriole. During this process, renin-expressing cells were distributed along the whole of the newly formed vessel. As the vessel matured, renin-expressing cells became restricted to the juxtaglomerular portion of the afferent arteriole. It is concluded that throughout life and within each individual arterial tree, expression of renin is heterogeneous, following patterns that are unique for each developmental stage. Furthermore, the association of renin-expressing cells with branching of renal arterioles suggests a role for these cells in the development of the kidney vasculature.

Evolution of chronic nitric oxide inhibition hypertension: relationship to renal function.

We conducted longitudinal measurements of blood pressure and renal function in the conscious, chronically catheterized rat before and during acute nitric oxide synthase inhibition (N-nitro-L-arginine methylester [L-NAME], 37 micromol/kg IV) and then chronic administration of oral L-NAME (approximately 37 micromol/kg per 24 hours). These studies specifically investigate the impact on plasma and renal renin as well as volume status during the evolution of this hypertension in rats not subjected to acute experimental stress. Blood pressure progressively increased with chronic administration of L-NAME and reached values greatly above those seen with acute administration of L-NAME. There were parallel increases in renal vascular resistance and development of proteinuria, and glomerular filtration rate began to decline at day 21, coincident with the appearance of renal damage. Twenty-four-hour urinary nitrite and nitrate excretion remained depressed, reflecting reduced nitric oxide synthesis. The plasma renin activity was variable and only increased transiently at 21 days, thus the angiotensin II dependence of this hypertension is not caused by stimulated plasma renin activity. Despite severe hypertension, sodium intake and excretion were unchanged over the 21 days of L-NAME administration. Plasma volume was significantly reduced at days 2 and 12 of L-NAME administration; thus the prolonged plasma volume contraction must result from the acute natriuretic response to the initial acute L-NAME administration.