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Clinical Science (2013) 124, 351370 (Printed in Great Britain) doi: 10.

1042/CS20120378

Insulin signalling to the kidney in health and disease


Lorna J. HALE and Richard J. M. COWARD
Academic and Childrens Renal Unit, University of Bristol, Learning and Research, Southmead Hospital, Bristol BS10 5NB, U.K.

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Abstract
Ninety-one years ago insulin was discovered, which was one of the most important medical discoveries in the past century, transforming the lives of millions of diabetic patients. Initially insulin was considered only important for rapid control of blood glucose by its action on a restricted number of tissues; however, it has now become clear that this hormone controls an array of cellular processes in many different tissues. The present review will focus on the role of insulin in the kidney in health and disease.
Key words: diabetes, diabetic nephropathy, insulin, intracellular signalling, kidney, metabolic syndrome

OVERVIEW OF CELLULAR INSULIN SIGNALLING


The role of insulin in the human body has been an active subject of interest since the discovery of insulin in 1921 by Banting, Best, Collip and Macleod. The critical importance of this nding was recognized by the Nobel Committee in 1923 when they awarded Banting and Macleod the Nobel Prize in Physiology or Medicine [1] just 2 years after their discovery. In the 50 years that followed the effects of insulin were intensely studied and revealed its glucose-controlling effects focusing on the liver, muscle and adipose tissue [2]. Insulin is a highly potent physiological anabolic hormone that promotes the synthesis and storage of lipids, carbohydrates and proteins, while also inhibiting their degradation and release back into the circulation. In mammals insulin is the main hormone controlling blood glucose; it achieves this by stimulating glucose inux and metabolism in muscles and adipocytes, and by inhibiting gluconeogenesis by the liver. These tissues have always been considered the classically insulin-sensitive organs of the body. However, insulin has the ability to modify the expression and/or activity of an assortment of enzymes and transport systems in a wide variety of cell types [3], as the present review will describe.

INSULIN AND IGF (INSULIN-LIKE GROWTH FACTOR) RECEPTORS


When discussing the receptors that insulin can signal through it is important to consider another closely related collection of hormones, namely the IGF family. The reason for this is that the IGF hormones, IGF-I and IGF-II, have structural similarity to insulin and their major functional receptor, IGF-IR (IGF-I receptor) [4,5], is also structurally similar to the IR (insulin receptor). The signicance of this is that insulin can signal through the IGFIR and likewise IGF-I/-II can signal via the IR, although with differing afnities. Indeed, it is even more complicated than this as hybrid receptors are formed, by combinations of the IR and IGF-IR, through which all of the hormones can signal but with differing afnities (Table 1). Insulin has the greatest afnity for the IR, so the rest of the present review will predominantly focus on this receptor. The IR in humans is located on chromosome 19 and is encoded by a gene containing 22 exons and 21 introns spanning 120 kb [6,7]. It is a heterotetrameric receptor consisting of two and two subunits [8], which are linked by disulde bonds in a --- conguration (Figure 1). The subunits are extracellular and have the insulin-binding domain, whereas the subunits

Clinical Science

, , Abbreviations: BK channel, large-conductance Ca2 + -activated K + channel; BP blood pressure; CAP Cbl-associated protein; DM, diabetes mellitus; DN, diabetic nephropathy; DOK, downstream of kinase; ECM, extracellular matrix; eNOS, endothelial NO synthase; ERK, extracellular-signal-regulated kinase; ESRD, end-stage renal disease; FSGS, focal segmental glomerulosclerosis; GBM, glomerular basement membrane; GFB, glomerular ltration barrier; GFR, glomerular ltration rate; GLUT, glucose transporter; GEnC, glomerular endothelial cell; Grb2, growth-factor-receptor-bound protein 2; IGF, insulin-like growth factor; IGF-IR, IGF-I receptor; IL, interleukin; IR, insulin receptor; IRS, insulin receptor substrate; ksp, kidney-specic; JNK, c-Jun N-terminal kinase; MAPK, mitogen-activated protein kinase; MPGN, membranoproliferative glomerulonephritis; mTOR, mammalian target of rapamycin; mTORC, mTOR complex; OMIM, Online Mendelian Inheritance in Man ; PH, pleckstrin homology; PI3K phosphoinositide 3-kinase; podIRKO mouse, podocyte-specic IR-decient transgenic mouse; PPAR , peroxisome-proliferator-activated receptor ; PTB, phosphotyrosine-binding; Raptor, regulatory associated protein of mTOR; RBF, renal blood ow; SGK1, serum- and glucocorticoid-induced protein kinase 1; SH2, Src homology 2; SOS, Son of Sevenless; TRPC, transient receptor potential cation channel; TSC, tuberous sclerosis complex. Correspondence: Dr Richard Coward (email Richard.Coward@bristol.ac.uk).

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Table 1

Receptor subtypes of the insulin/IGF system in mammals Variable assembly of receptors showing the primary ligand-binding afnities for each. Ligand(s) Receptor Homotetramers IR-B/IR-B IR-A/IR-A IGF-IR/IGF-IR Heterotetramers (hybrids) IR-B/IR-A IR-B/IGF-IR IR-A/IGF-IR HIR-AB HR-B HR-A Insulin and IGF-II IGF-I IGF-I, IGF-II and insulin IGF-II IR-B IR-A IGF-IR Insulin Insulin and IGF-II IGF-I and IGF-II IGF-II IGF-I IGF-I Insulin Name Level of afnity . . . High-level Mid-level Low-level

have three compartmental domains: extracellular, transmembrane and cytosolic domains. Tyrosine residues in the cytosolic domain of subunits are involved in signal transduction and are autophosphorylated when insulin binds to the receptor or by exogenous tyrosine kinase activity [8]. There is a further level of complexity within the IR as it exists in two different isoforms, A and B, which are formed due to the inclusion or exclusion of exon 11 of the IR gene [911]. IR-A lacks exon 11, whereas IR-B includes it. IR-A is widely expressed throughout the body but is importantly up-regulated during prenatal development and when cells become cancerous [7]. IR-B is expressed largely in the classically insulin-sensitive tissues of liver, skeletal muscle and adipose tissue. Interestingly IR-B is also expressed in the kidney [12,13]. The IR isoforms dimerize and can form either pure or hybrid receptors with each other or the IGF-IR. The receptor make-up dictates the afnity of the cell for insulin and/or the IGF ligands, as the different receptors have differing afnities for each of these molecules (Table 1). It should also be noted that IRs are not solely located in glucoseregulating insulin target tissues, but in many other tissue types, suggesting other functional roles of insulin signalling in multiple biological systems distinct from glucose homoeostasis. The IR and IGF-IR mediate the actions of IGF-I, IGF-II and insulin. The IGF-IR shares a high degree of homology with the IR [14,15] (Figure 1). It is therefore unsurprising that insulin is capable of activating the IGF-IR and vice versa. IGF-I has the greatest afnity for the IGF-IR, followed by IGF-II, with insulin having a 500-fold lower afnity in comparison with its primary ligands [14].

CELLULAR INSULIN SIGNALLING PATHWAYS


The majority of work in this eld has been performed on adipocytes, liver and skeletal muscle, as these are crucial for postprandial glucose regulation in response to insulin. The insulin signal transduction pathway is highly conserved and responsible for the regulation of a number of aspects of cellular physiology, most notable of which is the metabolic effects of glucose uptake and its utilization within the cell. Following a meal, increased levels of insulin encourage enhanced glucose uptake, metabolism and storage within muscle and adipose cells

[16]. Insulin levels rapidly increase approximately 10-fold after a meal from a basal level of approximately 50 pmol to 600 pmol [17]. GLUTs (glucose transporters) are energy-independent and allow glucose to enter or leave the cell, passively down a concentration gradient, when they are incorporated into the cell membrane. The classic insulin-responsive glucose transporter is GLUT4 [18], which translocates from a cytoplasmic vesicular pool to the plasma membrane in response to insulin. This is the signature molecule of rapidly insulin-sensitive cells that absorb glucose. However, there is also robust evidence that GLUT1 [19] can also translocate in a similar manner from an intracellular pool to the plasma membrane and rapidly increase its plasma membrane concentration in response to insulin. GLUT1 is also a constitutional transporter in many cells [20]. Here it sits at the plasma membrane of cells continuously and allows a constant delivery of glucose for cellular function. The IR differs from many other receptor tyrosine kinases in that, instead of recruiting downstream effector molecules to its phosphorylated cytoplasmic domains, when activated it phosphorylates a number of scaffolding proteins which then in turn are responsible for recruiting various downstream effector proteins [21]. A number of intracellular substrates have been discovered, including the IRS (insulin receptor substrate) family (IRS1 IRS4), IRS5/DOK4 (downstream of kinase 4), IRS/DOK5, Gab1, Cbl, APS [adaptor protein with PH (pleckstrin homology) and SH2 (Src homology 2) domains] and Shc isoforms, and SIRP (signal regulatory protein) family members [22,23]. The best characterized have been the IRS family of proteins [24]. IRS proteins do not possess intrinsic catalytic activities, and are instead composed of multiple interaction domains and phosphorylation motifs. Four IRS proteins have been identied (IRS1IRS4), with IRS1 and IRS2 being the most widely expressed. Each IRS protein has the distinct characteristics of an N-terminus PH domain adjacent to a PTB (phosphotyrosine-binding) domain, ending in a variable length C-terminus. The C-terminal tail of each IRS protein contains tyrosine phosphorylation sites that serve as on/off switches and recruit the downstream signalling proteins. IRS1 and IRS2 have the longest tail therefore providing them with a greater number of possible phosphorylation sites (20) in comparison with IRS3 and IRS4 [24]. The IRS proteins form an important node of control for the regulation of insulin and IGF signal transduction in cells.

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Figure 1

Structure of the IR and IGF-IR Both the IR and the IGF-IR are transmembrane tyrosine kinase cell-surface receptors that mediate the actions of IGF-I, IGF-II and insulin [5]. The IGF-IR shares a high degree of homology with the IR [14] and, like the IR, forms a --- tetramer composed of two and two subunits joined by disulde bonds [15]. This Figure was adapted and reprinted from Brain Research Reviews, 44(23), Hawkes, C., and Kar, S., The insulin-like growth factor-II/mannose-6-phosphate receptor: structure, distribution and function in the central nervous system, 117140, Copyright (2004), with permission from Elsevier.

When the intrinsic tyrosine kinase activity of the receptor is triggered by insulin binding, three major signalling pathways have been described that are propagated in response: (i) CAP (Cbl-associated protein), (ii) the PI3K (phosphoinositide 3-kinase) pathway, and (iii) the MAPK (mitogen-activated protein kinase) pathway (Figure 2). The PI3K pathway is one of the best characterized downstream effectors of the IRS proteins and activates many of the metabolic functions of insulin. Association of the p85 regulatory subunit with IRS proteins leads to the activation of further downstream molecules, including Akt substrates (which form another functional node in the pathway) and mTOR (mammalian target of rapamycin) [25,26], eventually resulting in PI3K being targeted to the plasma membrane [22]. There is evidence that mTOR is important for kidney function and this will be discussed in more detail later in the present review. The stimulation of glucose uptake by insulin is mediated by PI3K-dependent and PI3K-independent pathways, which play a vital role in the translocation of GLUT4. This is highlighted by use of the PI3K inhibitor wortmannin, which is able to completely block the uptake of glucose into cells upon insulin stimulation [27]. However, despite the critical role of PI3K in insulinstimulated glucose uptake, activation of at least a second pathway distinct from PI3K is also necessary [28]. This is evident from a number of studies that have examined different elements of the pathway during insulin signalling. By overexpressing a constitutively active membrane-bound form of Akt in 3T3L1 adipo-

cytes, glucose transport and GLUT4 translocation increases in the absence of insulin [29]; conversely, the insulin-stimulated translocation of GLUT4 is inhibited by the expression of a dominant-negative Akt mutant [30]. These results indicate that Akt is required for insulin signalling. However, if the PI3K pathway is activated by factors other than insulin, such as PDGF (platelet-derived growth factor) or IL (interleukin)-4, although these factors can robustly activate PI3K and Akt, they do not possess insulins ability to stimulate GLUT4 translocation and glucose uptake [28]. This suggests that other pathways also need to be activated by insulin to elicit an effect. A number of studies have suggested that a separate signalling pathway exists for the IR in microdomains within the cell, such as lipid rafts [22]. It has been proposed that insulin can also activate the GTPase TC10, via lipid-raft localization of the CAPCblCrk complex and the guanine-nucleotide-exchange factor C3G [31], and initiate glucose uptake in cells. This process occurs independently of PI3K and has been shown to be crucial to insulin-stimulated GLUT4 translocation [32,33]. However, in contrast with this, ndings by Mitra et al. [34] have reported that Cbl/CAP isoforms are in fact not required components of insulin signalling to GLUT4 transporters; therefore the precise role of this pathway remains contentious. The nal pathway through which insulin can act is the MAPK pathway. This pathway controls a range of cellular activities, including differentiation, proliferation, transformation, survival and death [3537]. The mammalian MAPK family consists of

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Figure 2

Substrates of the IR family and the major cellular signalling pathways evoked There are a number of critical nodes which regulate the biological response to each stimulus. These include the CAP/Cbl pathway (1), the PI3K pathway (2) via Akt and the MAPKs (3). AS160, Akt substrate of 160 kDa; FOXO1, forkhead box O1; GSK-3, glycogen synthase-3; PIP2, phosphatidylinositol 4,5-bisphosphate; PIP3, phosphatidylinositol 3,4,5-trisphosphate; PDK1, phosphoinositide-dependent kinase 1; PKB, protein kinase B; PTP1B, protein tyrosine phosphatase 1B; pY, tyrosine phosphorylation.

p38, ERK (extracellular-signal-regulated kinase) and JNK (c-Jun N-terminal kinase), each of which exist in a number of isoforms: p38-, -, - and , ERK18 and JNK13 [35,38]. In terms of insulin signalling, the MAPK pathway is activated following the binding of Grb2 (growth-factor-receptor-bound protein 2) and SOS (Son of Sevenless) to phosphotyrosine residues on Shc and Gab1 [39]. Phosphorylation of certain tyrosine residues on Gab1 are required for binding to and activation of the protein tyrosine phosphatase SHP2 (SH2 domain-containing protein tyrosine phosphatase 2) [40,41], whereas phosphorylation of certain tyrosine residues on Shc allow the binding of Grb2/SOS to these sites [42]. This binding initiates activation of the GTPase Ras, followed shortly by Raf, leading to a kinase cascade resulting in the phosphorylation and activation of the MAPK pathway [39]. It is important to note that the p85/p110 PI3K complex also binds to Ras, thereby connecting two pathways which are often considered to be separate [43]. With regard to insulin signalling, the MAPK pathway is primarily associated with the regulation of mitogenesis [44]. Again, stimulation of the MAPK pathway in isolation is not able to induce glucose uptake in fat or muscle.

In summary, insulin transduces its signal through at least three different cellular pathways. There are also a number of critical nodes involved in insulin signalling, including the IRIGF-IR complexes, IRS molecules and Akt/MAPK substrates that are able to modify the biological effects of a ligand on a particular cell type. What is currently unclear is how cells are able to control which pathways are activated in response to specic ligands. This could be of great therapeutic benet as it may enable novel ways of overcoming cellular resistance or hyperstimulation of either the insulin or IGF pathways.

INSULIN SIGNALLING IS NOT ALL ABOUT GLUCOSE HOMOEOSTASIS


IR, IGF-IR and hybrid receptors are expressed throughout the body in most tissues and are not restricted to the classic insulinsensitive glucose uptake tissues, such as liver, muscle and fat [45]. It is therefore not surprising that insulin has other

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important biological effects on many different tissues independent of glucose uptake, as evolution rarely allows redundant systems to persist. Two good examples of important non-glucosecontrolling actions of insulin are found in the brain and the cardiovascular system. In the brain, insulin signalling is crucial for controlling appetite, regulating food intake [4648] and preventing obesity and infertility [45]. These are all features associated with insulin resistance. Similarly, within the cardiovascular system, insulin can directly signal to both the heart dynamically regulating metabolism [49] and also the peripheral vasculature controlling tone. Cardiovascular tone is controlled both centrally [50] and through local modulation of endothelial NO release by insulin [51]. This is potentially relevant in the cardiovascular morbidity and mortality associated with insulin-resistant states.

INSULIN AND THE KIDNEY


The kidney is the major organ that regulates uid balance, BP (blood pressure), acidbase status, haemoglobin production, electrolyte control and waste removal in the body. It is a highly vascular structure that receives 20 % of the circulating blood volume per min through the renal artery: the RBF (renal blood ow). From here, glomeruli within the kidney are perfused and their ltrate per min is dened as the GFR (glomerular ltration rate). Within each kidney are approximately 1 million nephrons. These are the functional units of the kidney. Each nephron has a primary ltering unit, the glomerulus, and a tubule, which is able to modify the primary ltrate produced from the glomeruli. There is now accumulating evidence that both the glomeruli and tubules are insulin sensitive.

glucose 6-phosphate by glucose-6-phosphatase. Originally, it was thought that the liver was the only organ in the body that could release glucose in times of starvation and stress. However, it is now clear that the kidneys are also capable of gluconeogenesis. This explains why patients with fulminant liver failure can maintain their circulating glucose concentrations [61]. It transpires that both the liver and the kidney possess sufcient gluconeogenic enzyme and glucose 6-phosphate activity to facilitate this production [62]. Furthermore, the kidney not only releases glucose in times of acidosis or after prolonged fasting as initially thought, but is also able to release signicant amounts of glucose in normal post-absorptive individuals [63]. Insulin reduces renal gluconeogenesis [64,65], so is an important factor in the control of glucose release from here. The importance of renal gluconeogenesis is discussed in depth in a review by Gerich et al. [62] and highlights that the kidney may be equally as important in the production and regulation of gluconeogenesis as the liver. The cellular location of gluconeogenesis in the kidney is not entirely clear; however, the proximal tubules have been shown to express glucose-6-phosphatase as have the parietal cells and podocytes [66]. Clinically, this may be relevant as congenital loss of glucose-6-phosphatase causes glycogen storage disease type I. These children have developmental delay, episodes of hypoglycaemia and, from a renal perspective, proximal renal tubular dysfunction and glomerular damage as illustrated by FSGS (focal segmental glomerulosclerosis) on renal biopsy [67]. Therefore, interestingly, the renal phenotype of this condition revolves around the two cell types that are known to express glucose-6phosphatase in the kidney.

Haemodynamic control of renal organ blood ow and local glomerular blood ow: the role of insulin
There are conicting ndings on the direct role of insulin in RBF through the major vessels and local glomerular blood ow within the kidney, which appears to be species-dependent. Early work on conscious dogs suggested that insulin decreased RBF and that this was independent of the effects of insulin on the sympathetic nervous system [68]. However, in humans, a number of studies [6972], but not all [73,74], have shown that RBF actually increases in response to insulin. It is possible that the differences reported in these studies are due to dual actions of insulin. Insulin is able to act systemically by stimulating a catecholamine response and activating the sympathetic nervous system [75]. However, it can also cause a local renal vasodilatory effect in the kidney. A more consistent nding is that insulin increases the GFR in insulin-sensitive subjects. This occurs through local renal vasodilation and is mediated by a prostoglandin-dependent pathway that can be blocked with indomethacin [76] and regulated by eNOS (endothelial NO synthase) [77]. Interestingly, a number of groups have shown that insulin increases the GFR in normal subjects, but this response is lost in insulin-resistant subjects [72,78,79]. However, and in contrast, a recent study has identied a human polymorphism in the IRS1 gene which is associated with an increased GFR [80], and the authors speculate that loss of renal insulin signalling may, in fact, be responsible for an increased GFR.

INSULIN AND THE KIDNEY: IN HEALTH


The active insulin molecule, after loss of the c-peptide, exists as a monomer; it contains 51 amino acids and is approximately 6 kDa in size, allowing it to freely traverse the GFB (glomerular ltration barrier) and pass into the tubular lumen [6]. This is important as it allows insulin rapid access to all of the cells in the glomerulus and nephron after it has been secreted into the circulation and passes through the renal artery. The major high-afnity receptor for insulin, the IR, is located throughout the kidney in all of the cells of the glomerulus [5255] and the entire length of the renal tubule, from the proximal tubule to the collecting ducts [5659]. Recently, it has also been shown that kidney, as a whole, abundantly expresses IR isoform B [60], which is the isoform found in the classically insulin-responsive, glucose-regulating, tissues of fat, skeletal muscle and liver. It is now also clear that insulin is involved in a number of homoeostatic physiological responses throughout the kidney and are described below.

Renal gluconeogenesis
Two main processes, glycogenolysis and gluconeogenesis, result in the release of glucose, both of which involve the hydrolysis of

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Figure 4 Transmission electron micrograph of the glomerular capillary cell wall Three interacting layers make up the glomerular capillary cell wall: GEnCs, whose fenestrations are denoted by , the GBM, and podocytes, whose tertiary foot processes are denoted by FP Examples of a . slit diaphragm between the foot processes are indicated by the thick arrows.

primary urine across the GFB during an average human lifetime and, as the primary urine is practically protein-free, this means that more than 200 000 kg of albumin has to be prevented from crossing [81].

Podocytes
Podocytes are unique cells found on the urinary side of the GFB (Figure 4). In the past 15 years, it has become clear that they are critically important in maintaining the integrity of the GFB and preventing leakage of albumin into the urine. There are now more than ten inherited human genetic mutations, all of which cause nephrotic syndrome and all of which code for proteins found predominantly in the podocyte (Table 2). Podocytes are embryonically derived from mesenchyme and are classied by most as epithelial cells, as they are polarized and sit on a basement membrane. However, they also have features of other cell types, including smooth muscle, as they are contractile and express smooth muscle markers [82], and neurons due to their processes, secretory capacity and, for the most part, inability to replicate when fully formed [83]. Podocytes adhere to the GBM through a network of anchoring proteins [8486] and have specialized modied adherens junctions, called slit diaphragms, formed between their foot processes. These contain a set of proteins that are crucial in maintaining the integrity of the GFB [81,87]. It is now clear that the podocyte depends on its actin cytoskeleton to maintain its structure and the integrity of the GFB and many of the slit diaphragm proteins are linked to this [88,89]. Although the majority of disease-causing mutations in the podocyte are related to actin-regulating functions, it is of note that gain-of-function mutations in the Ca2 + channel modulating TRPC6 [TRPC

Figure 3 The glomerulus The diagram in the upper panel is a simplied view of the glomerulus. This is shown in elegant detail using electron microscopy (lower panel), where the specic cell types and distinct areas of the glomerulus are highlighted in a mouse glomerulus from our laboratory. The upper panel was reproduced from Postgraduate Medical Journal, Vinen, C.S., and Oliveira, D.B, 79, 206213, 2003 with permission from BMJ Publishing Group Ltd.

Specic cellular actions of insulin throughout the nephron Glomerulus


Glomeruli are composed of three different resident cell types: podocytes, GEnCs (glomerular endothelial cells) and mesangial cells. It is now clear that all of these cells respond to insulin, but in different ways. Podocytes and GEnCs are separated by the GBM (glomerular basement membrane) and constitute the GFB. Mesangial cells are specialized smooth-muscle-like cells that are able to contract and regulate blood ow to the glomerulus (Figure 3). The glomeruli lter as much as 5 million litres of

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Table 2

Podocyte mutations associated with disease Protein(s) encoded Nephrin Structurally related protein to nephrin -Actinin-4 Wilms tumour protein LIM homeobox transcription factor 1- CD2-associated protein Podocin Transient receptor potential cation channel, subfamily C, member 6 Formin Phospholipase C, 1 Co-enzyme Q10 biosynthesis mono-oxygenase 6 Protocadherin Fat 1 Consequence(s) of mutation Massive proteinuria in utero, nephrosis at birth: congenital nephrotic syndrome of the Finnish type (NPHS1) [202] Proteinuria and perinatal lethality in mice at 8 weeks [203] Results in autosomal-domininant form of FSGS; adult onset nephrotic syndrome [204] Linked to DenysDrash syndrome, pseudohermaphroditism; proteinuria occurs early [205] Childhood-onset proteinuria, regulates expression of multiple podocyte genes required for differentiation and function; causes nail-patella syndrome [206] Congenital nephrotic syndrome [207] Steroid-resistant nephrotic syndrome; presents in early childhood [208] Autosomal-dominant FSGS; normally presents in late childhood/early adulthood [90] Autosomal-dominant FSGS; adolescence or adulthood presentation: often mild proteinuria initially [209] Early onset nephrotic syndrome with ESRD [93] Early onset nephrotic syndrome, sensorineural deafness and early lethality in some [92] Proteinuria and perinatal lethality in mice [210]

Gene names NPHS1 NEPH1 ACTN4 WT1 LMX1B CD2AP NHPS2 TRPC6 INF2 PLCE1 COQ6 FAT

(transient receptor potential cation channel), subfamily C, member 6] has also been discovered to cause nephrotic syndrome [90,91], as well as mitochondrial proteins [92] and enzymatic proteins [PLCE1 (phospholipase C, 1)] [93]. In recent years, our group [52,94] and others [95] have shown that the podocyte is a rapidly insulin-responsive cell. Our initial work employed a conditionally immortalized human podocyte cell line [96] to study insulin responses in this cell. This was helpful as the cell line contains a temperature-sensitive transgene that enables the cells to replicate and proliferate ad innitum at 33 C but when they are thermo-switched to 37 C they exit the cell cycle and are able to differentiate and express many of the markers of maturity. This is important as the thermo-switched cells resemble mature podocytes found in the normal glomerulus. We found that differentiated podocytes rapidly respond to insulin by doubling their glucose uptake within 15 min [97]. This is similar to the kinetics observed in muscle, which is not surprising given the muscle-like features and markers that podocytes exhibit [82]. Importantly proliferative immature 33 C human podocytes, human proximal tubular cells and human GEnCs did not respond to insulin in respect to glucose uptake. We went on to show that this process was dependent on the actin cytoskeleton and activated translocation of the glucose transporters GLUT1 and GLUT4 from cytoplasmic vesicles to the plasma membrane of the cell [97]. We extended these observations to show that the podocyte protein nephrin was also important in insulin signalling in the podocyte [98]. This was achieved by studying conditionally immortalized podocytes derived from children with the most severe form of congenital nephrotic syndrome called Finnish type congenital nephrotic syndrome. This occurs secondary to mutations in the protein nephrin. We developed a number of conditionally immortalized natural human knockout cell lines from nephrindecient (no nephrin protein made) or nephrin-mutant (protein

made but unable to target to the plasma membrane) kidneys. We found that these cells were completely unresponsive to insulin in respect to glucose uptake, but could be rescued by genetically reconstituting nephrin back into them. Mechanistically this was due to a failure of GLUT-rich vesicles to dock and become incorporated into the plasma membrane of the nephrin-decient/mutant podocytes. We went on to show that the C-terminus of nephrin was able to form a proteinprotein association with VAMP2 (vesicle-associated membrane 2), which is important for vesicle docking with the plasma membrane through SNARE (soluble Nethylmaleimide-sensitive fusion protein-attachment protein receptor) processes [98]. Although nephrin is only expressed in the podocyte in the kidney it is also found in pancreatic -cells in the body, where it has also been shown to be potentially involved in insulin release here in response to glucose [99]. Interestingly given the possible role of nephrin in both insulin release and its cellular action, children with nephrin mutations who receive kidney transplants do not appear to develop overt DM (diabetes mellitus). This suggests that, although nephrin may be involved in the control of insulin release, it is not critical. Recently, we have generated a podocyte-specic IR-decient transgenic (podIRKO) mouse, which has proven to be highly informative [94]. podIRKO mice develop albuminuria and loss of foot process architecture by 8 weeks of age. In light of this, we have also found that insulin is able to rapidly remodel the lamentous actin cytoskeleton of podocytes. This is via the IR and through modulation of small GTPases, which act as molecular switches for actin remodelling in cells. RhoA is activated and CDC42 is inhibited [94]. We have also shown that in the podocyte insulin stimulates the PI3K and MAPK signalling cascades, but not the CAP/Cbl pathway. Another recent exciting advance in our understanding of the homoeostatic role of insulin on podocyte biology has come from Dryers group. They have discovered that insulin rapidly increases

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Figure 5

Post-prandial actions of insulin in the podocyte Insulin is released into the circulation by the pancreas and then reaches the podocyte by passing through the GFB. It is small (6 kDa), so freely passes through here. It then causes the podocyte changes depicted in a short time frame of less than 15 min.

the membrane expression of Ca2 + and K + channels in the podocyte [100]. They have demonstrated that, within minutes, insulin causes TRPC6 to increase on the surface of podocytes, but TRPC5 channels decrease. Functionally this causes an increased inux of cations, particularly Ca2 + , into the cell. This elegant study went on to show that the effects of insulin on TRPC6 were mediated through the production of ROS (reactive oxygen species) via activation of NADPH oxidases [100]. The same group have also shown that, in addition to modulating TRPC channels in podocytes, insulin also rapidly causes Ca2 + -regulated K + channels to locate to the plasma membrane of this cell [101]. Collectively these ndings suggest that, when insulin stimulates the podocyte, as occurs after a meal, it causes the podocyte to rapidly take up a readily usable energy source, glucose, remodel its actin cytoskeleton and contract, which is facilitated by ionic ux into the cell. We think this makes biological sense in order for this cell to brace itself for the increased work this cell is required to perform at this time (Figure 5).

GEnCs
GEnCs line the capillaries of the glomerulus (Figure 4). They are highly fenestrated and therefore freely permeable to an array of molecules. These cells express IRs [55,102], IGF-IRs [103] and hybrid receptors [104], so have the receptor apparatus for insulin signalling. Early work has suggested that their primary role was to remove insulin from the circulation and degrade it [105]. However, we have shown that insulin activates the PI3K pathway rapidly in human GEnCs [94], but it does not have the same biological actions in these cells as in podocytes and does not cause rapid glucose uptake [52] or actin remodelling [94]. Using human GEnCs that have been conditionally immortalized

[106] in the same way as podocytes, we have found that insulin rapidly induces the production of eNOS (J. Hurcombe and R.J.M. Coward, unpublished work). This complements work by Mima et al. [107], who have demonstrated that glomeruli isolated from rats are insulin-sensitive and insulin also rapidly induces eNOS. Recently, it has been shown that reduced endothelial insulin signalling elsewhere in the body reduces local eNOS production and impairs the physical delivery of insulin to extra-capillary sites [108]. As insulin is secreted into the circulation from the pancreatic -cells, it needs to traverse the endothelial layer of blood vessels to reach extra-vascular targets. However, in the glomerulus, the GEnC is highly fenestrated, which may protect the underlying podocytes from insulin deciency when the GEnC is insulin-resistant. Elucidating the functional importance of insulin signalling in the GEnC in the intact glomerulus is currently challenging. This is partly due to a lack of transgenic tools to be able to genetically manipulate the GEnC. Specically, there is currently no way of targeting cre recombinase to this cell within the kidney, which is important when developing cell-specic-knockout mice models using cre-loxp technology [109]. However, there is now promise that this may be rectied in the future, as specic GEnC genes have recently been identied within the kidney which may be benecial in developing these mice [110] and genetically manipulating the GEnC in the intact glomerulus in the future.

Mesangial cells
Mesangial cells are specialized contractile cells that support the glomerular capillaries. Their contractile properties show similarities to vascular smooth muscle cells [111,112], with the release of Ca2 + from stores within the ER (endoplasmic reticulum) upon the initiation of contraction. The released Ca2 + activates Cl

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channels, which then depolarize the cell membrane and activate VOCCs (voltage-operated Ca2 + channels), resulting in an increase in intracellular Ca2 + levels and the subsequent activation of BK (large-conductance Ca2 + -activated K + ) channels. Activation of BK channels then causes the cell membrane potential to hyperpolarize [113,114]. Both the IR and IGF-IR are expressed in mesangial cells, although at differing levels, with the IGF-IR being predominant [54]. Despite this, insulin at high levels is able to stimulate mesangial cell proliferation through activation of either receptor [115]. BK channels are expressed in abundance in mesangial cells, where they contribute to the relaxation of the cell and as a result an increased GFR [116]. Similar to podocytes, insulin has been shown to increase the density of BK channels in the plasma membrane of human mesangial cells via activation of the MAPK pathway [117]. Unlike the podocyte, mesangial cells do not rapidly increase glucose in response to insulin and, as a result, the intracellular glucose level observed more directly reects its plasma concentration. Excessive extracelluar glucose in the diabetic mileu can enter mesangial cells with ease via GLUT1 in an insulinindependent manner, which can result in glucotoxicity. High glucose levels have been shown to enhance GLUT1 expression in mesangial cells, which may result in progressive damage [118]; in addition, these glucose-induced effects can be mimicked in normal glucose conditions by specically overexpressing GLUT1 [119], resulting in excessive production of ECM (extracellular matrix) proteins [120]. Apoptosis is an important mechanism within a number of organs, tissues and cells, and the glomerulus is no exception. Apoptosis of glomerular cells is a closely regulated process; it can be benecial in allowing the removal of excess cells in order to resolve glomerular injury, but can also be detrimental if excessive apoptosis is allowed to occur leading to hypocellularity [121]. Insulin has been shown to be a pro-survival factor via activation of PI3K, which in turn allows the recruitment of Akt to the plasma membrane, where it phosphorylates a number of molecules to suppress apoptosis [122,123]. Both insulin and IGF-I protect mesangial cells from a variety of apoptotic triggers via the PI3K/Akt pathway using this mechanism [124]; in conjunction with these ndings, insulin has also been shown to reduce ERK1/2 activation and increase levels of the cyclin-dependent kinase inhibitor p21 during apoptosis, providing an additional level of protection.

The tubule consists of a number of different specialized polarized epithelial cells that are able to transport molecules to and from the tubular lumen into and from the circulation. Numerous groups have shown that the IR, IGF-IR and hybrid receptors are expressed throughout the tubule [56,59,125,126], including the proximal tubule, loop of Henle, distal convoluted tubule and collecting ducts. More insulin binds to the tubules in comparison with the glomeruli, although with less afnity as reported by some [127], but not all groups [57]. In our initial work, we studied immortalized proximal tubular cells (HK2) and found that they did not respond to insulin in respect to glucose uptake [52]. However, it is clear that the tubules are insulin-responsive, but that insulin elicits different cellular effects in this part of the kidney. Elegant work by Mima et al. [107] has shown that ex vivo renal tubules from rats are insulin-responsive in respect to activation of the PI3K and MAPK pathways. Interestingly, this insulin response is not lost in established diabetes, as was found with insulin responses in the glomeruli in these studies. Furthermore, in vitro studies of isolated tubular cell types suggest that insulin can rapidly modify a variety of transporter systems throughout the tubule [128131]. These include the NHE3 (Na + /H + exchanger type III) [132,133], which is the major Na + transporter in the proximal tubule and responsible for 65 % of Na + resorption here. Modulation of this channel is also able to alter the acidbase status in the body. Insulin also augments Na + resorption through other transporters throughout the tubules, including the loop of Henle, via the butamide-sensitive Na + K + 2Cl channels [131], and ENaC (epithelial Na + channel), Na + /K + ATPase and recently the Na + Cl co-transporter [134] in the collecting ducts. A number of studies examining the distal tubule have demonstrated that insulin binds to the IR and activates the PI3K pathway, which then phosphorylates and stimulates SGK1 (serum- and glucocorticoid-induced protein kinase 1) phosphorylation [135] that inhibits the breakdown of transporters through endocytic retrieval pathways and may also directly phosphorylate transporters resulting in enhanced actions [136]. This is interesting, as SGK1 seems to be a connection through which mineralocorticoids and insulin can modify Na + retention in the distal part of the nephron. In addition to Na + , insulin can also modulate the resorption of other ions in the proximal tubule, including PO4 3 (phosphate ion) through the Na + PO4 3 cotransporter type-II in the proximal tubule [137] and Mg2 + in the distal convoluted tubules [138]. A comprehensive review of the role of insulin in the renal tubules was performed in 2007 and we would recommend reading this excellent article [139].

Renal tubules
These are segregated into a number of dened regions, including the proximal tubule, loop of Henle, distal convoluted tubule and the collecting ducts. This part of the kidney is able to modulate the primary ltrate from the glomerulus by reabsorbing ions back into the blood or secreting ions into the urine. Important ions that are regulated include glucose, Na + and HCO3 resorption in the proximal tubule, Na + and water resorption in the loop of Henle, and K + and H + secretion in the distal convoluted tubule, together with water retention in the collecting ducts. Collectively, the tubules are able to control acidbase status, Na + and water resorption and hence BP regulation in the body.

INSULIN SIGNALLING AND THE KIDNEY: DISEASE Systemic insulin-resistant states


Insulin-resistant states are a major global healthcare problem in the 21st century, with an estimated 171 million diabetics present in the world. However, owing to current sedentary lifestyles, population aging and urbanization, the anticipated number of cases is predicted to more than double in the next 15 years [140]. The incidence of the insulin-resistant metabolic syndrome is even

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Figure 6

Histological changes in the kidney from a patient with DM These images demonstrate the characteristic changes observed in the glomeruli of patients with DN compared with those of a healthy patient. Upper panels show a normal glomerulus. Light microscopy Periodic AcidSchiff staining (left-hand panel), PAAg (Periodic Acid silver) staining (middle panel) and transmission electron microscopy (TEM) at 2900 magnication (right-hand panel). Lower panels show the classic features of DN. Glomerulosclerosis, mesangial matrix, mesangial hypercellularity and KimmelsteinWilson lesions (arrowed) are shown in the light microscopy pictures. Thickening of basement membrane are shown in the transmission electron microscopy pictures on the right. These pictures were generated and supplied by Dr Tibor Toth, Department of Pathology, Southmead Hospital, Bristol, U.K.

more pronounced, with an estimated prevalence of 20 % in the people over 20 years of age in the U.S.A. rising to over 40 % when over 60 years of age being reported [141]. Approximately one-third of all new cases of ESRD (end-stage renal disease) worldwide is accounted for by DM [142] and, in the U.S.A., this is even higher with over 50 % of new patients having DM (U.S. Renal Data System, USRDS 2011 Annual Data Report; http://www.usrds.org). Type 2 DM is due to insulin resistance of peripheral tissues, in contrast with Type 1 DM which that occurs secondary to insulin deciency caused by destruction of the cells of the pancreas [143]. DN (diabetic nephropathy) is the most common microvascular chronic complication of DM [144]. DN is a progressive disease which takes several years to develop; it occurs in 3040 % of patients with Type 1 DM and 810 % of patients with Type 2 DM [87]. Its natural history is dominated by progressive albuminuria. There is now accumulating evidence that a loss of insulin responses in the kidney may contribute to a number of the complications that occur in insulin-resistance states, including albuminuric glomerular disease and hypertension.

The glomerulus, insulin, diabetes and the metabolic syndrome


Early renal manifestations of DN are focused on the glomerulus in the kidney consisting of glomerular hyperltration and microalbuminuria, alongside other changes, including GBM thickening, mesangial expansion and accumulation of ECM proteins such as laminin, collagen and bronectin [144]. Advanced DN is characterized by increased albuminuria (macroalbumin-

uria), glomerulosclerosis, interstitial brosis and ESRD [87,144] (Figure 6). In recent years, the podocyte has become an intense focus of research into this eld as loss of this cell has been found to be the best histological predictor of progression in DN [145]. Furthermore, as progressive albuminuria dominates the natural history of DN, this also makes the podocyte an attractive target cell in DN, because of its crucial role in preventing albuminuria, as discussed above (Table 2) [146]. The metabolic syndrome is also associated with microalbuminuria; indeed, it is part of the diagnostic criteria in some classications, including that of the World Health Organization. As described above, we have shown that the human podocyte is an insulin-sensitive cell [52] and we have developed podIRKO mice. These mice were highly informative as they developed a number of features of DN, including albuminuria, glomerulosclerosis, matrix accumulation (including type-IV collagen), thickening of the GBM and podocyte apoptosis. However, they all had normal blood glucose control, demonstrating that none of these features were driven by hyperglycaemia. This suggests that insulin signalling to the podocyte is critically important for normal glomerular function and may also have a role in some aspects of DN. It should be noted that the podIRKO mice only exhibited some features of DN and did not have enlarged kidneys nor did they have mesangial hypercellularity or the classic nodular KimmelsteinWilson lesions of DN (Figure 6). A potential explanation for this is that in diabetes there is a loss of insulin sensitivity of the podocyte and this results in some of the pathological consequences associated with DN, but other aspects of DN, for example renal hypertrophy and mesangial expansion,

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are driven by other factors. These may include pathways driven by high glucose levels or other growth factors such as IGF-I or IGF-II acting on the cells of the glomerulus. Our current working hypothesis is that, in both Type 1 and Type 2 DM, as well as the metabolic syndrome, in those patients who develop nephropathy, there is insulin resistance in the podocyte which contributes to the development of renal damage. Type 2 DM and the metabolic syndrome are intrinsic cellular insulin-resistant conditions, so this hypothesis would appear intuitively to be correct. Type 1 DM occurs due to a lack of insulin; however, there is compelling evidence that those patients with Type 1 DM who develop nephropathy are more likely to also be insulin-resistant. Prolonged Type 1 DM causes insulin resistance [147], and nephropathic compared with non-nephropathic patients with Type 1 DM require larger doses of insulin to control their diabetes [148], are more insulin-resistant when assessed by euglycaemic clamps [149] and are more likely to have a strong family history of cellular insulin resistance [150]. There is further experimental evidence to support this hypothesis. Tejada et al. [95] have examined podocyte insulin responses in the development of albuminuria in the db/db Type 2 DM mouse model and have shown that IR and PI3K signalling is lost early in the disease process. Furthermore, a recent excellent rodent study [107] examining Type 1 and Type 2 DM models has examined the effect of diabetes on insulin signalling in the kidney. Mima et al. [107] studied rats given streptozotocin inducing Type 1 DM and the Zucker obese model of Type 2 DM. They allowed the rats to develop diabetes and then examined their insulin signalling pathways in the glomerular and tubular compartments of the kidney. They found that insulin rapidly initiated PI3K and MAPK signalling in both isolated glomeruli and tubular fractions of control rats. However, when the rats had either Type 1 or Type 2 DM they both resulted in a loss of insulin signalling via the PI3K pathway specically in the glomerular and not in the tubular compartment. They went on to show that high glucose increased ubiquitination and hence loss of IRS1 in the glomeruli, which could be a mechanistic pathway through which Type 1 DM is able to directly modulate cellular insulin signalling in the glomerulus. Finally, building on previous work [151,152], they demonstrated that inhibiting PKC (protein kinase C) was able to reverse high-glucose-induced insulin resistance in GEnCs via eNOS. This adds weight to the assumption that GEnC as well as podocyte insulin resistance may be important in the development of glomerular complications in systemic insulin-resistant states. A consequence of increased insulin signalling in the podocyte is increased translocation of GLUT4 and GLUT1 to the plasma membrane of this cell, allowing more glucose to passively diffuse into the cell. Previously, it has been proposed that a major reason for cellular dysfunction in the setting of diabetes is glucose toxicity of cells [153]. In the glomerulus there is evidence that overexpression of GLUTs in the mesangial cell is detrimental to function [154]. However, this does not seem to be the case for podocytes. An elegant study by the Brosius group [155] has shown that increasing the glucose transporter GLUT1 specically in the podocyte in a model of Type 2 DM is not detrimental to glomerular function, but intriguingly seems to be protective against the development of some aspects of DN. GLUT1 is a

glucose transporter that is expressed at the cell surface constitutionally in many cells and allows basal glucose uptake, but is also found in insulin-responsive translocatable glucose transporter vesicular pools similar to GLUT4. One possibility why this mouse was protected from the development of DN is that it may have been protected from episodes of glucose deciency when stimulated by insulin [155]. Our research has also shown that nephrin is crucial for the insulin sensitivity of podocytes. Many groups have demonstrated that nephrin is reduced early in DN [156158]. Potentially this could be inducing insulin resistance in these cells resulting in pathological changes. However, it is also possible that the loss of nephrin in the development of DN is also a consequence of podocyte dysfunction and not a cause. What is currently unclear is the in vivo insulin responsiveness of the podocyte in systemic insulin-resistant states. Some groups have found that, similar to the classically metabolically insulin-responsive tissues of adipose [159], liver [160] and skeletal muscle [161], that the IR is down-regulated at the protein level in the kidneys of models of Type 2 DM and systemic insulin resistance. However, other groups have found in Type 2 DM that, although adipose, liver and skeletal muscle demonstrate diminished insulin binding and signalling, the kidney does not [57,162,163]. Therefore it is possible that early in the development of diabetes there is hyperstimulation of the insulin signalling axis and not loss of insulin sensitivity in the kidney, and this may also be having a detrimental effect. There is evidence that insulin can modulate the GFB, which may be through direct insulin ligand receptor binding. Approximately 30 years ago insulin-decient human subjects with Type 1 DM were given insulin under euglycaemic clamp conditions, i.e. maintaining the blood glucose constant, and this resulted in a transient increase in albumin excretion into the urine [164]. Furthermore, it was shown that giving a glucose load to healthy subjects and also patients with Type 1 DM only caused an increase in urinary albumin excretion in the healthy subjects [165]. This suggests that hyperglycaemia is not the major driver causing a loss of albumin into the urine, as this occurred in the patients with Type 1 DM who did not develop albuminuria (and could not produce insulin in response to the glucose challenge), but that in healthy subjects high glucose levels stimulated insulin release (which lowers their blood glucose levels) and caused albumin to leak into the urine. These insulin effects may explain the immediate post-prandial proteinuria that is widely described in human subjects and rodents [166,167]. If podocyte insulin resistance is a key factor in the development of nephropathy in diabetes then it follows that strategies to enhance podocyte cellular insulin sensitivity could be benecial in treating this condition. There is evidence that some agents which have insulin-sensitizing properties, including metformin and PPAR (peroxisome-proliferator-activated receptor ) agonists, are benecial in preventing kidney damage in models of DN in both Type 1 [168] and Type 2 [169,170] DM, as well as other non-diabetic chronic kidney diseases [171,172], which partially supports this premise. Indeed, we have shown that the PPAR agonist rosiglitazone is able to directly enhance insulin sensitivity of the podocyte in vitro [173].

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Figure 7

Insulin regulates the mTOR complex and is also regulated by it This simplied diagram demonstrates how insulin is able to activate mTORC1. It is now also clear that inhibiting mTOR chronically with rapamycin can induce insulin resistance in cells. This is through a loss of mTORC2, which causes decreased Akt signalling. MEK, MAPK/ERK kinase; PTEN, phosphatase and tensin homologue deleted on chromosome 10; Rictor, rapamycin-insensitive companion of mTOR; RSK, ribosomal S6 kinase.

Conversely, we have found that factors that are increased systemically in the insulin-resistant metabolic syndrome render the podocyte insulin-resistant. These include non-esteried free fatty acids such as palmitate [174]. Other groups have also explored some of the molecular inhibitors of insulin signalling in the podocyte. An elegant study demonstrated that SHIP2 (SH2domain-containing inositol phosphatase) was able to inhibit insulin signalling in the podocyte and that it was up regulated in the glomeruli in a rat model of Type 2 DM [175]. Discovering these molecules is important, as they could potentially be good therapeutic targets when inhibited. Finally, there are ndings indicating that insulin can regulate mesangial cell function in the glomerulus and when this cell is rendered resistant it potentially contributes to matrix formation [176], mesangial expansion [177] and hyperltration [178]. In summary, there is now evidence that all three cell types found in the glomerulus can respond to insulin, but they respond in different ways. Furthermore, when insulin signalling is altered in the glomerulus it results in pathology.

mTOR, insulin and glomerular disease: another important pathway


Another potentially important role for insulin in the glomerulus is modulating the mTOR pathway. The mTOR signalling cascade controls cellular protein synthesis, growth, metabolism, autophagy and survival in response to growth factors, stress, energy and nutrient stimuli. mTOR is a protein kinase and the catalytic subunit of two functional complexes: mTORC1 (mTOR complex 1) and mTORC2 (mTOR complex 2) [179]. mTORC1 is a rapamycin-sensitive complex in which mTOR is associated with the Raptor (regulatory associated protein of mTOR) and regulates a number of cellular

processes, including protein synthesis, cell growth and proliferation [179,180]. Insulin is able to increase the activity of mTORC1 through PI3K and ERK1/2 pathways via TSC2 (tuberous sclerosis complex 2). Both pathways inhibit TSC2, which then prevents it from suppressing mTORC1 expression. This results in protein translation, ribosomal biogenesis and autophagy [181]. Interestingly, it has also recently become evident that mTOR inhibition causes cellular insulin resistance [182] and that this action is through the mTORC2 complex [183]. Therefore the mTOR pathway is both controlled by, and also controls, insulin signalling (Figure 7). Rapamycin (Sirolimus) is used in renal transplantation because inhibiting mTOR inhibits the response of B- and T-cells to IL-2 and hence prevents organ rejection. mTOR expression is low or undetectable in the normal kidney but following ischaemia/reperfusion injury it increases signicantly, presumably to enable cellular repair and regeneration to occur. In this setting, rapamycin is detrimental as it inhibits mTOR and results in delayed renal repair and recovery [184]. The function of mTOR in the glomerulus remains controversial; however, the ndings from a number of recent elegant murine transgenic studies [185187] have advanced the understanding of the role of mTOR in one cell type located in the kidney, namely the podocyte. Previous work has reported that systemic administration of rapamycin in mouse models of both Type 1 and Type 2 DM can prevent the progression of DN [188 190], suggesting it may be clinically benecial to inhibit mTOR in these settings. However, it has also been shown that mTOR inhibition with rapamycin can be detrimental in non-diabetic conditions and can cause proteinuria and glomerulosclerosis in both humans and rodents [191194]. Studies published recently by Inoki et al. [185], G del et al. [186] and Cina et al. [187] have highlighted the o

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critical importance of the mTOR pathway in podocyte biology. By manipulating mTORC1 and mTORC2 they have shown that loss of mTORC1 is detrimental to glomerular function [186] and this is exacerbated further when mTORC2 is also lost [186,187]. Interestingly increasing the activity of mTORC1 can also be harmful to the glomerulus causing albuminuric renal disease resembling DN [185]. Increased mTORC1 activity occurs in DN and can be rescued by genetic or pharmacological inhibition of mTORC1. In summary, insulin signalling can modify mTOR activity in the glomerulus, predominantly through mTORC1 but it can also be modied and suppressed when mTORC2 is inhibited by long-term rapamycin therapy.

The tubule, insulin, diabetes and the metabolic syndrome


As discussed above, insulin is able to modulate tubular function in a number of ways. One of the most important effects seems to be its action on Na + resorption from the tubule into the circulation. As hypertension is a prominent feature of the metabolic syndrome and DN, it is not surprising that a great deal of work has explored the potential role of tubular insulin sensitivity and its role in the control of BP. In 2008, Tiwari et al. [195] developed a mouse model in which the IR was knocked out in the renal tubules by crossing a oxed IR mouse with a ksp (kidney-specic)-cadeherin promoter-linked cre recombinase-producing mouse. Ksp-cadeherin is only found in the tubular epithelial cells in the kidney. It is predominantly expressed in the distal aspect of the tubules from the thick ascending loop of Henle through to the collecting ducts [196]. As discussed in the previous section, work based predominantly on cell culture models had suggested that a loss of tubular insulin signalling would result in a reduction of Na + resorption from the urinary ltrate and hence a naturesis with an associated lowering of the BP. However, intriguingly, this was not the case with this model. These mice had impaired urinary Na + excretion in response to a Na + load and were hypertensive in comparison with controls. The group went on to show that insulin signalling here was able to activate local NO production and reduce BP in wildtype normal animals, presumably through a vasodilatory effect, but in tubular IR-knockout mice this did not occur. This suggests another role of tubular insulin signalling in NO production and BP control. It may also be clinically relevant as hypertension is commonly associated with insulin-resistant states.

tions such as skeletal muscle and liver. These patients are often severely insulin-resistant at a cellular level. The most severe form of lipodystophy is the generalized form where the patients have no adipose tissue. The majority of patients with congenital and acquired forms of this condition have glomerular disease with albuminuria, but only a small subset have the classical features of DN. The rest have a variety of renal pathologies, including FSGS and MPGN (membranoproliferative glomerulonephritis). Another cohort of patients suffer from acquired partial lipodystrophies. These patients lose adipose tissue from their face, neck, upper extremities, thorax and upper abdomen, and have immunological abnormalities with low C3 complement levels and elevated C3 nephritic factor levels. The most common renal lesion found in these patients is MPGN type 2 (dense deposit disease). It has been hypothesized, but not categorically proven, that this is an immunological disease; however, there may be a contribution from insulin resistance. The renal phenotypes of extreme insulin resistance are extensively reviewed by Musso et al. [197], which we would recommend reading.

Diseases that target the IR Auto-antibodies against the IR (type B insulin resistance).
This is associated with extreme insulin resistance and DM in the majority, although paradoxically these auto-antibodies can also cause hypoglycaemia in some situations [198]. Patients usually have an underlying collagen vascular disease, most commonly systemic lupus erythromatosis. Again, more than 50 % of patients have albuminuria but their histology is normally that of one of the forms of lupus nephritis [198,199].

Mutations of the IR (type A insulin resistance). Two syndromes account for the majority of patients suffering from mutations of the IR: Donogue syndrome, previously known as Leprechaunism [OMIM (Online Mendelian Inheritance in Man ) 147670], and RabsonMendenhall syndrome (OMIM 262190). These patients have extreme insulin resistance, acanthosis nigrans, hirsuitism and are generally slender. These patients commonly, but not always, have DN [200], which may be secondary to the cellular insulin signalling defect they experience.

FUTURE DIRECTIONS
It is now clear that the kidney is an insulin-sensitive organ, but that different regions of the kidney respond to insulin in different ways. To elucidate the clinical relevance of insulin sensitivity of the kidney there are still some fundamental questions that need to be addressed and will be a focus of research in the upcoming years. These questions include the following. (i) Does insulin resistance in the GEnC and/or mesangial cell contribute to glomerular pathology? (ii) Do human renal cells in vivo become insulin-resistant in systemic insulin-resistant states? This has never been proven. (iii) Can we therapeutically manipulate the insulin signalling pathway in the kidney to prevent renal disease from developing? Ideally this would be kidney (cell)-specic to reduce off-target side effects, as have been experienced with

Rare renal disease associated with severe insulin resistance


In addition to the very common conditions of DM and the metabolic syndrome there are a number of rare syndromic forms of cellular insulin resistance that are associated with glomerular renal disease. It is interesting that these do not always result in classic DN, but rather in a spectrum of renal pathologies as follows.

Lipodystrophies
This group of disorders is caused by a failure to deposit fat in adipose tissues. This causes abnormalities in circulating adipokines and results in the deposition of fat in ectopic loca-

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other insulin-sensitizing drugs, such as the glitazones [201]. (iv) Why does insulin elicit different biological responses in different tissues? This is probably due to differences in important signalling nodes in the insulin and IGF pathways, but again this is not proven.

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CONCLUSIONS
It is now clear that the kidney is an insulin-responsive organ in a variety of different ways. Manipulating these responses may have great therapeutic potential in treating glomerular disease and hypertension associated with DM and other insulin-resistant states.
ACKNOWLEDGMENT

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We thank Dr Tibor Toth (Department of Pathology, Southmead Hospital, Bristol, U.K.) for producing Figure 6.

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Our own work was supported by Kidney Research UK and the Medical Research Council.

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REFERENCES
1 Banting, F. G., Best, C. H., Collip, J. B., Campbell, W. R. and Fletcher, A. A. (1922) Pancreatic extracts in the treatment of diabetes mellitus. Can. Med. Assoc. J. 12, 141146 White, M. F. (2003) Insulin signaling in health and disease. Science 302, 17101711 White, M. F. and Kahn, C. R. (1994) The insulin signaling system. J. Biol. Chem. 269, 14 Megyesi, K., Kahn, C. R., Roth, J., Neville, Jr, D. M., Nissley, S. P Humbel, R. E. and Froesch, E. R. (1975) The NSILA-s ., receptor in liver plasma membranes. Characterization and comparison with the insulin receptor. J. Biol. Chem. 250, 89908996 Kim, J. J. and Accili, D. (2002) Signalling through IGF-I and insulin receptors: where is the specicity? Growth Horm. IGF Res. 12, 8490 De Meyts, P (2004) Insulin and its receptor: structure, function . and evolution. BioEssays 26, 13511362 Belore, A., Frasca, F., Pandini, G., Sciacca, L. and Vigneri, R. (2009) Insulin receptor isoforms and insulin receptor/insulin-like growth factor receptor hybrids in physiology and disease. Endocr. Rev. 30, 586623 Lee, J. and Pilch, P F. (1994) The insulin receptor: structure, . function and signaling. Am. J. Physiol. 266, C319C334 Kasuga, M., Zick, Y., Blithe, D. L., Crettaz, M. and Kahn, C. R. (1982) Insulin stimulates tyrosine phosphorylation of the insulin receptor in a cell-free system. Nature 298, 667669 Ebina, Y., Ellis, L., Jarnagin, K., Edery, M., Graf, L., Clauser, E., Ou, J. H., Masiarz, F., Kan, Y. W., Goldne, I. D. et al. (1985) The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling. Cell 40, 747758 Ullrich, A., Bell, J. R., Chen, E. Y., Herrera, R., Petruzzelli, L. M., Dull, T. J., Gray, A., Coussens, L., Liao, Y. C., Tsubokawa, M. et al. (1985) Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature 313, 756761

20

21

2 3 4

22

23

24 25 26

6 7

27

8 9

28

10

29

11

30

Bailyes, E. M., Nave, B. T., Soos, M. A., Orr, S. R., Hayward, A. C. and Siddle, K. (1997) Insulin receptor/IGF-I receptor hybrids are widely distributed in mammalian tissues: quantication of individual receptor species by selective immunoprecipitation and immunoblotting. Biochem. J. 327, 209215 Moller, D. E., Yokota, A., Caro, J. F. and Flier, J. S. (1989) Tissue-specic expression of two alternatively spliced insulin receptor mRNAs in man. Mol. Endocrinol. 3, 12631269 Samani, A. A., Yakar, S., LeRoith, D. and Brodt, P (2007) The . role of the IGF system in cancer growth and metastasis: overview and recent insights. Endocr. Rev. 28, 2047 Ullrich, A., Gray, A., Tam, A. W., Yang-Feng, T., Tsubokawa, M., Collins, C., Henzel, W., Le Bon, T., Kathuria, S., Chen, E. et al. (1986) Insulin-like growth factor I receptor primary structure: comparison with insulin receptor suggests structural determinants that dene functional specicity. EMBO J. 5, 25032512 Kahn, B. B. (1992) Facilitative glucose transporters: regulatory mechanisms and dysregulation in diabetes. J. Clin. Invest. 89, 13671374 Gannon, M. C., Nuttall, J. A. and Nuttall, F. Q. (2002) Oral arginine does not stimulate an increase in insulin concentration but delays glucose disposal. Am. J. Clin. Nutr. 76, 10161022 Pessin, J. E., Thurmond, D. C., Elmendorf, J. S., Coker, K. J. and Okada, S. (1999) Molecular basis of insulin-stimulated GLUT4 vesicle trafcking. Location! Location! Location! J. Biol. Chem. 274, 25932596 Clarke, J. F., Young, P W., Yonezawa, K., Kasuga, M. and . Holman, G. D. (1994) Inhibition of the translocation of GLUT1 and GLUT4 in 3T3-L1 cells by the phosphatidylinositol 3-kinase inhibitor, wortmannin. Biochem. J. 300, 631635 Olson, A. L. and Pessin, J. E. (1996) Structure, function and regulation of the mammalian facilitative glucose transporter gene family. Annu. Rev. Nutr. 16, 235256 Saltiel, A. R. and Kahn, C. R. (2001) Insulin signalling and the regulation of glucose and lipid metabolism. Nature 414, 799806 Chang, L., Chiang, S. H. and Saltiel, A. R. (2004) Insulin signaling and the regulation of glucose transport. Mol. Med. 10, 6571 Watson, R. T., Kanzaki, M. and Pessin, J. E. (2004) Regulated membrane trafcking of the insulin-responsive glucose transporter 4 in adipocytes. Endocr. Rev. 25, 177204 White, M. F. (2002) IRS proteins and the common path to diabetes. Am. J. Physiol. Endocrinol. Metab. 283, E413E422 Cantley, L. C. (2002) The phosphoinositide 3-kinase pathway. Science 296, 16551657 Wang, Q., Somwar, R., Bilan, P J., Liu, Z., Jin, J., Woodgett, J. R. . and Klip, A. (1999) Protein kinase B/Akt participates in GLUT4 translocation by insulin in L6 myoblasts. Mol. Cell. Biol. 19, 40084018 Okada, T., Kawano, Y., Sakakibara, T., Hazeki, O. and Ui, M. (1994) Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adipocytes. Studies with a selective inhibitor wortmannin. J. Biol. Chem. 269, 35683573 Isakoff, S. J., Taha, C., Rose, E., Marcusohn, J., Klip, A. and Skolnik, E. Y. (1995) The inability of phosphatidylinositol 3-kinase activation to stimulate GLUT4 translocation indicates additional signaling pathways are required for insulin-stimulated glucose uptake. Proc. Natl. Acad. Sci. U.S.A. 92, 1024710251 Kohn, A. D., Summers, S. A., Birnbaum, M. J. and Roth, R. A. (1996) Expression of a constitutively active Akt Ser/Thr kinase in 3T3-L1 adipocytes stimulates glucose uptake and glucose transporter 4 translocation. J. Biol. Chem. 271, 3137231378 Cong, L. N., Chen, H., Li, Y., Zhou, L., McGibbon, M. A., Taylor, S. I. and Quon, M. J. (1997) Physiological role of Akt in insulin-stimulated translocation of GLUT4 in transfected rat adipose cells. Mol. Endocrinol. 11, 18811890

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48

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Saltiel, A. R. and Pessin, J. E. (2002) Insulin signaling pathways in time and space. Trends Cell Biol. 12, 6571 Baumann, C. A., Ribon, V., Kanzaki, M., Thurmond, D. C., Mora, S., Shigematsu, S., Bickel, P E., Pessin, J. E. and Saltiel, A. R. . (2000) CAP denes a second signalling pathway required for insulin-stimulated glucose transport. Nature 407, 202207 Chiang, S. H., Baumann, C. A., Kanzaki, M., Thurmond, D. C., Watson, R. T., Neudauer, C. L., Macara, I. G., Pessin, J. E. and Saltiel, A. R. (2001) Insulin-stimulated GLUT4 translocation requires the CAP-dependent activation of TC10. Nature 410, 944948 Mitra, P Zheng, X. and Czech, M. P (2004) RNAi-based analysis ., . of CAP Cbl and CrkII function in the regulation of GLUT4 by , insulin. J. Biol. Chem. 279, 3743137435 Dhillon, A. S., Hagan, S., Rath, O. and Kolch, W. (2007) MAP kinase signalling pathways in cancer. Oncogene 26, 32793290 McCubrey, J. A., Lahair, M. M. and Franklin, R. A. (2006) Reactive oxygen species-induced activation of the MAP kinase signaling pathways. Antioxid. Redox Signaling 8, 17751789 Torii, S., Yamamoto, T., Tsuchiya, Y. and Nishida, E. (2006) ERK MAP kinase in G cell cycle progression and cancer. Cancer Sci. 97, 697702 Schaeffer, H. J. and Weber, M. J. (1999) Mitogen-activated protein kinases: specic messages from ubiquitous messengers. Mol. Cell. Biol. 19, 24352444 Taniguchi, C. M., Emanuelli, B. and Kahn, C. R. (2006) Critical nodes in signalling pathways: insights into insulin action. Nat. Rev. Mol. Cell Biol. 7, 8596 Rocchi, S., Tartare-Deckert, S., Murdaca, J., Holgado-Madruga, M., Wong, A. J. and Van Obberghen, E. (1998) Determination of Gab1 (Grb2-associated binder-1) interaction with insulin receptor-signaling molecules. Mol. Endocrinol. 12, 914923 Cunnick, J. M., Mei, L., Doupnik, C. A. and Wu, J. (2001) Phosphotyrosines 627 and 659 of Gab1 constitute a bisphosphoryl tyrosine-based activation motif (BTAM) conferring binding and activation of SHP2. J. Biol. Chem. 276, 2438024387 Salcini, A. E., McGlade, J., Pelicci, G., Nicoletti, I., Pawson, T. and Pelicci, P G. (1994) Formation of Shc-Grb2 complexes is . necessary to induce neoplastic transformation by overexpression of Shc proteins. Oncogene 9, 28272836 Rodriguez-Viciana, P Warne, P H., Dhand, R., Vanhaesebroeck, ., . B., Gout, I., Fry, M. J., Watereld, M. D. and Downward, J. (1994) Phosphatidylinositol-3-OH kinase as a direct target of Ras. Nature 370, 527532 Denton, R. M. and Tavare, J. M. (1995) Does mitogen-activatedprotein kinase have a role in insulin action? The cases for and against. Eur. J. Biochem. 227, 597611 Bruning, J. C., Gautam, D., Burks, D. J., Gillette, J., Schubert, M., Orban, P C., Klein, R., Krone, W., Muller-Wieland, D. and . Kahn, C. R. (2000) Role of brain insulin receptor in control of body weight and reproduction. Science 289, 21222125 Baskin, D. G., Figlewicz Lattemann, D., Seeley, R. J., Woods, S. C., Porte, Jr, D. and Schwartz, M. W. (1999) Insulin and leptin: dual adiposity signals to the brain for the regulation of food intake and body weight. Brain Res. 848, 114123 Schwartz, M. W., Figlewicz, D. P Baskin, D. G., Woods, S. C. ., and Porte, Jr, D. (1992) Insulin in the brain: a hormonal regulator of energy balance. Endocr. Rev. 13, 387414 Schwartz, M. W., Sipols, A. J., Marks, J. L., Sanacora, G., White, J. D., Scheurink, A., Kahn, S. E., Baskin, D. G., Woods, S. C., Figlewicz, D. P et al. (1992) Inhibition of hypothalamic . neuropeptide Y gene expression by insulin. Endocrinology 130, 36083616 Bertrand, L., Horman, S., Beauloye, C. and Vanoverschelde, J. L. (2008) Insulin signalling in the heart. Cardiovasc. Res. 79, 238248

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

Cabou, C., Cani, P D., Campistron, G., Knauf, C., Mathieu, C., . Sartori, C., Amar, J., Scherrer, U. and Burcelin, R. (2007) Central insulin regulates heart rate and arterial blood ow: an endothelial nitric oxide synthase-dependent mechanism altered during diabetes. Diabetes 56, 28722877 Zeng, G. and Quon, M. J. (1996) Insulin-stimulated production of nitric oxide is inhibited by wortmannin. Direct measurement in vascular endothelial cells. J. Clin. Invest. 98, 894898 Coward, R. J., Welsh, G. I., Yang, J., Tasman, C., Lennon, R., Koziell, A., Satchell, S., Holman, G. D., Kerjaschki, D., Tavare, J. M. et al. (2005) The human glomerular podocyte is a novel target for insulin action. Diabetes 54, 30953102 Arnqvist, H. J., Ballermann, B. J. and King, G. L. (1988) Receptors for and effects of insulin and IGF-I in rat glomerular mesangial cells. Am. J. Physiol. 254, C411C416 Conti, F. G., Striker, L. J., Lesniak, M. A., MacKay, K., Roth, J. and Striker, G. E. (1988) Studies on binding and mitogenic effect of insulin and insulin-like growth factor I in glomerular mesangial cells. Endocrinology 122, 27882795 Elliot, S. J., Conti, F. G., Striker, L. J. and Striker, G. E. (1990) Mouse glomerular endothelial cells have an insulin receptor. Horm. Metab. Res. 22, 557560 Butlen, D., Vadrot, S., Roseau, S. and Morel, F. (1988) Insulin receptors along the rat nephron: [125 I] insulin binding in microdissected glomeruli and tubules. Pugers Arch. 412, 604612 Sechi, L. A., De Carli, S. and Bartoli, E. (1994) In situ characterization of renal insulin receptors in the rat. J. Recept. Res. 14, 347356 Bourdeau, J. E., Chen, E. R. and Carone, F. A. (1973) Insulin uptake in the renal proximal tubule. Am. J. Physiol. 225, 13991404 Nakamura, R., Emmanouel, D. S. and Katz, A. I. (1983) Insulin binding sites in various segments of the rabbit nephron. J. Clin. Invest. 72, 388392 Vienberg, S. G., Bouman, S. D., Sorensen, H., Stidsen, C. E., Kjeldsen, T., Glendorf, T., Sorensen, A. R., Olsen, G. S. andersen, B. and Nishimura, E. (2011) Receptor-isoform-selective insulin analogues give tissue-preferential effects. Biochem. J. 440, 301308 Battezzati, A., Caumo, A., Martino, F., Sereni, L. P Coppa, J., ., Romito, R., Ammatuna, M., Regalia, E., Matthews, D. E., Mazzaferro, V. et al. (2004) Nonhepatic glucose production in humans. Am. J. Physiol. Endocrinol. Metab. 286, E129E135 Gerich, J. E., Meyer, C., Woerle, H. J. and Stumvoll, M. (2001) Renal gluconeogenesis: its importance in human glucose homeostasis. Diabetes Care 24, 382391 Meyer, C., Dostou, J. M., Welle, S. L. and Gerich, J. E. (2002) Role of human liver, kidney and skeletal muscle in postprandial glucose homeostasis. Am. J. Physiol. Endocrinol. Metab. 282, E419E427 Cersosimo, E., Garlick, P and Ferretti, J. (1999) Insulin . regulation of renal glucose metabolism in humans. Am. J. Physiol. 276, E78E84 Meyer, C., Dostou, J., Nadkarni, V. and Gerich, J. (1998) Effects of physiological hyperinsulinemia on systemic, renal and hepatic substrate metabolism. Am. J. Physiol. 275, F915F921 Burchell, A. and Hume, R. (1995) The glucose-6-phosphatase system in human development. Histol. Histopathol. 10, 979993 Martens, D. H., Rake, J. P Navis, G., Fidler, V., van Dael, C. M. ., and Smit, G. P (2009) Renal function in glycogen storage . disease type I, natural course and renopreservative effects of ACE inhibition. Clin. J. Am. Soc. Nephrol. 4, 17411746 Liang, C., Doherty, J. U., Faillace, R., Maekawa, K., Arnold, S., Gavras, H. and Hood, Jr, W. B. (1982) Insulin infusion in conscious dogs. Effects on systemic and coronary hemodynamics, regional blood ows and plasma catecholamines. J. Clin. Invest. 69, 13211336

www.clinsci.org

365

L. J. Hale and R. J. M. Coward

69

70

71

72

73

74

75

76

77

78

79

80

81 82

83

84

Stenvinkel, P Bolinder, J. and Alvestrand, A. (1992) Effects of ., insulin on renal haemodynamics and the proximal and distal tubular sodium handling in healthy subjects. Diabetologia 35, 10421048 Stenvinkel, P Ottosson-Seeberger, A. and Alvestrand, A. (1995) ., Renal hemodynamics and sodium handling in moderate renal insufciency: the role of insulin resistance and dyslipidemia. J. Am. Soc. Nephrol. 5, 17511760 Herlitz, H., Widgren, B., Urbanavicius, V., Attvall, S. and Persson, B. (1996) Stimulatory effect of insulin on tubular sodium reabsorption in normotensive subjects with a positive family history of hypertension. Nephrol. Dial. Transplant. 11, 4754 Ter Maaten, J. C., Bakker, S. J., Serne, E. H., Moshage, H. J., Donker, A. J. and Gans, R. O. (2000) Insulin-mediated increases in renal plasma ow are impaired in insulin-resistant normal subjects. Eur. J. Clin. Invest. 30, 10901098 DeFronzo, R. A., Cooke, C. R. andres, R., Faloona, G. R. and Davis, P J. (1975) The effect of insulin on renal handling of . sodium, potassium, calcium and phosphate in man. J. Clin. Invest. 55, 845855 Vierhapper, H., Gasic, S., Roden, M. and Waldhausl, W. (1993) Increase in skeletal muscle blood ow but not in renal blood ow during euglycemic hyperinsulinemia in man. Horm. Metab. Res. 25, 438441 Ward, K. D., Sparrow, D., Landsberg, L., Young, J. B., Vokonas, P S. and Weiss, S. T. (1996) Inuence of insulin, sympathetic . nervous system activity and obesity on blood pressure: the Normative Aging Study. J. Hypertens. 14, 301308 Cohen, A. J., McCarthy, D. M. and Stoff, J. S. (1989) Direct hemodynamic effect of insulin in the isolated perfused kidney. Am. J. Physiol. 257, F580F585 Hayashi, K., Fujiwara, K., Oka, K., Nagahama, T., Matsuda, H. and Saruta, T. (1997) Effects of insulin on rat renal microvessels: studies in the isolated perfused hydronephrotic kidney. Kidney Int. 51, 15071513 Tucker, B. J., Mendonca, M. M. and Blantz, R. C. (1993) Contrasting effects of acute insulin infusion on renal function in awake nondiabetic and diabetic rats. J. Am. Soc. Nephrol. 3, 16861693 ter Maaten, J. C., Bakker, S. J., Serne, E. H., ter Wee, P M., . Donker, A. J. and Gans, R. O. (1999) Insulins acute effects on glomerular ltration rate correlate with insulin sensitivity whereas insulins acute effects on proximal tubular sodium reabsorption correlation with salt sensitivity in normal subjects. Nephrol. Dial. Transplant. 14, 23572363 Thameem, F., Puppala, S., Schneider, J., Bhandari, B., Arya, R., Arar, N. H., Vasylyeva, T. L., Farook, V. S., Fowler, S., Almasy, L. et al. (2012) The Gly972 Arg variant of human insulin receptor substrate 1 gene is associated with variation in glomerular ltration rate likely through impaired insulin receptor signaling. Diabetes 61, 23852393 Simons, M. and Huber, T. B. (2008) Its not all about nephrin. Kidney Int. 73, 671673 Saleem, M. A., Zavadil, J., Bailly, M., McGee, K., Witherden, I. R., Pavenstadt, H., Hsu, H., Sanday, J., Satchell, S. C., Lennon, R. et al. (2008) The molecular and functional phenotype of glomerular podocytes reveals key features of contractile smooth muscle cells. Am. J. Physiol. Renal Physiol. 295, F959F970 Kobayashi, N., Gao, S. Y., Chen, J., Saito, K., Miyawaki, K., Li, C. Y., Pan, L., Saito, S., Terashita, T. and Matsuda, S. (2004) Process formation of the renal glomerular podocyte: is there common molecular machinery for processes of podocytes and neurons? Anat. Sci. Int. 79, 110 Sachs, N., Claessen, N., Aten, J., Kreft, M., Teske, G. J., Koeman, A., Zuurbier, C. J., Janssen, H. and Sonnenberg, A. (2012) Blood pressure inuences end-stage renal disease of Cd151 knockout mice. J. Clin. Invest. 122, 348358

85

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88 89

90

91

92

93

94

95

96

97

98

99

100

101

Pozzi, A. and Zent, R. (2012) Hold tight or youll fall off: CD151 helps podocytes stick in high-pressure situations. J. Clin. Invest. 122, 1316 Kretzler, M. (2002) Regulation of adhesive interaction between podocytes and glomerular basement membrane. Microsc. Res. Tech. 57, 247253 Reddy, G. R., Kotlyarevska, K., Ransom, R. F. and Menon, R. K. (2008) The podocyte and diabetes mellitus: is the podocyte the key to the origins of diabetic nephropathy? Curr. Opin. Nephrol. Hypertens. 17, 3236 Pavenstadt, H., Kriz, W. and Kretzler, M. (2003) Cell biology of the glomerular podocyte. Physiol. Rev. 83, 253307 Welsh, G. I. and Saleem, M. A. (2011) The podocyte cytoskeleton-key to a functioning glomerulus in health and disease. Nat. Rev. Nephrol. 8, 1421 Winn, M. P Conlon, P J., Lynn, K. L., Farrington, M. K., Creazzo, ., . T., Hawkins, A. F., Daskalakis, N., Kwan, S. Y., Ebersviller, S., Burchette, J. L. et al. (2005) A mutation in the TRPC6 cation channel causes familial focal segmental glomerulosclerosis. Science 308, 18011804 Reiser, J., Polu, K. R., Moller, C. C., Kenlan, P Altintas, M. M., ., Wei, C., Faul, C., Herbert, S., Villegas, I., Avila-Casado, C. et al. (2005) TRPC6 is a glomerular slit diaphragm-associated channel required for normal renal function. Nat. Genet. 37, 739744 Heeringa, S. F., Chernin, G., Chaki, M., Zhou, W., Sloan, A. J., Ji, Z., Xie, L. X., Salviati, L., Hurd, T. W., Vega-Warner, V. et al. (2011) COQ6 mutations in human patients produce nephrotic syndrome with sensorineural deafness. J. Clin. Invest. 121, 20132024 Hinkes, B., Wiggins, R. C., Gbadegesin, R., Vlangos, C. N., Seelow, D., Nurnberg, G., Garg, P Verma, R., Chaib, H., ., Hoskins, B. E. et al. (2006) Positional cloning uncovers mutations in PLCE1 responsible for a nephrotic syndrome variant that may be reversible. Nat. Genet. 38, 13971405 Welsh, G. I., Hale, L. J., Eremina, V., Jeansson, M., Maezawa, Y., Lennon, R., Pons, D. A., Owen, R. J., Satchell, S. C., Miles, M. J. et al. (2010) Insulin signaling to the glomerular podocyte is critical for normal kidney function. Cell Metab. 12, 329340 Tejada, T., Catanuto, P Ijaz, A., Santos, J. V., Xia, X., Sanchez, ., P Sanabria, N., Lenz, O., Elliot, S. J. and Fornoni, A. (2008) ., Failure to phosphorylate AKT in podocytes from mice with early diabetic nephropathy promotes cell death. Kidney Int. 73, 13851393 Saleem, M. A., OHare, M. J., Reiser, J., Coward, R. J., Inward, C. D., Farren, T., Xing, C. Y., Ni, L., Mathieson, P W. and Mundel, . P (2002) A conditionally immortalized human podocyte cell line . demonstrating nephrin and podocin expression. J. Am. Soc. Nephrol. 13, 630638 Coward, R. J., Welsh, G. I., Yang, J., Tasman, C., Lennon, R., Koziell, A., Satchell, S., Holman, G. D., Kerjaschki, D., Tavare, J. M. et al. (2005) The human glomerular podocyte is a novel target for insulin action. Diabetes 54, 30953102 Coward, R. J., Welsh, G. I., Koziell, A., Hussain, S., Lennon, R., Ni, L., Tavare, J. M., Mathieson, P W. and Saleem, M. A. (2007) . Nephrin is critical for the action of insulin on human glomerular podocytes. Diabetes 56, 11271135 Fornoni, A., Jeon, J., Varona Santos, J., Cobianchi, L., Jauregui, A., Inverardi, L., Mandic, S. A., Bark, C., Johnson, K., McNamara, G. et al. (2010) Nephrin is expressed on the surface of insulin vesicles and facilitates glucose-stimulated insulin release. Diabetes 59, 190199 Kim, E. Y. anderson, M. and Dryer, S. E. (2011) Insulin increases surface expression of TRPC6 channels in podocytes: Role of NADPH oxidases and reactive oxygen species. Am. J. Physiol. Renal Physiol. 302, F298F307 Kim, E. Y. and Dryer, S. E. (2011) Effects of insulin and high glucose on mobilization of slo1 BKCa channels in podocytes. J. Cell Physiol. 226, 23072315

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Bar, R. S., Hoak, J. C. and Peacock, M. L. (1978) Insulin receptors in human endothelial cells: identication and characterization. J. Clin. Endocrinol. Metab. 47, 699702 Conti, F. G., Elliot, S. J., Striker, L. J. and Striker, G. E. (1989) Binding of insulin-like growth factor-I by glomerular endothelial and epithelial cells: further evidence for IGF-I action in the renal glomerulus. Biochem. Biophys. Res. Commun. 163, 952958 Nitert, M. D., Chisalita, S. I., Olsson, K., Bornfeldt, K. E. and Arnqvist, H. J. (2005) IGF-I/insulin hybrid receptors in human endothelial cells. Mol. Cell. Endocrinol. 229, 3137 Rabkin, R., Tsao, T., Elliot, S. J., Striker, L. J. and Striker, G. E. (1993) Insulin uptake and processing by cultured mouse glomerular endothelial cells. Am. J. Physiol. 265, C453C459 Satchell, S. C., Tasman, C. H., Singh, A., Ni, L., Geelen, J., von Ruhland, C. J., OHare, M, J., Saleem, M. A., van den Heuvel, L. P and Mathieson, P W. (2006) Conditionally immortalized . . human glomerular endothelial cells expressing fenestrations in response to VEGF. Kidney Int. 69, 16331640 Mima, A., Ohshiro, Y., Kitada, M., Matsumoto, M., Geraldes, P ., Li, C., Li, Q., White, G. S., Cahill, C., Rask-Madsen, C. et al. (2011) Glomerular-specic protein kinase C--induced insulin receptor substrate-1 dysfunction and insulin resistance in rat models of diabetes and obesity. Kidney Int. 79, 883896 Kubota, T., Kubota, N., Kumagai, H., Yamaguchi, S., Kozono, H., Takahashi, T., Inoue, M., Itoh, S., Takamoto, I., Sasako, T. et al. (2011) Impaired insulin signaling in endothelial cells reduces insulin-induced glucose uptake by skeletal muscle. Cell Metab. 13, 294307 Sauer, B. (1998) Inducible gene targeting in mice using the Cre/lox system. Methods 14, 381392 Patrakka, J., Xiao, Z., Nukui, M., Takemoto, M., He, L., Oddsson, A., Perisic, L., Kaukinen, A., Szigyarto, C. A., Uhlen, M. et al. (2007) Expression and subcellular distribution of novel glomerulus-associated proteins dendrin, ehd3, sh2d4a, plekhh2 and 2310066E14Rik. J. Am. Soc. Nephrol. 18, 689697 Abrass, C. K. (1995) Diabetic nephropathy. Mechanisms of mesangial matrix expansion. West J. Med. 162, 318321 Kreisberg, J. I. and Ayo, S. H. (1993) The glomerular mesangium in diabetes mellitus. Kidney Int. 43, 109113 Okuda, T., Yamashita, N. and Kurokawa, K. (1986) Angiotensin II and vasopressin stimulate calcium-activated chloride conductance in rat mesangial cells. J. Clin. Invest. 78, 14431448 Wang, X., Pluznick, J. L., Settles, D. C. and Sansom, S. C. (2007) Association of VASP with TRPC4 in PKG-mediated inhibition of the store-operated calcium response in mesangial cells. Am. J. Physiol. Renal Physiol. 293, F1768F1776 Soos, M. A. and Siddle, K. (1989) Immunological relationships between receptors for insulin and insulin-like growth factor I: evidence for structural heterogeneity of insulin-like growth factor I receptors involving hybrids with insulin receptors. Biochem. J. 263, 553563 Dalla Vestra, M., Saller, A., Mauer, M. and Fioretto, P (2001) . Role of mesangial expansion in the pathogenesis of diabetic nephropathy. J. Nephrol. 14 (Suppl. 4), S51S57 Foutz, R. M., Grimm, P R. and Sansom, S. C. (2008) Insulin . increases the activity of mesangial BK channels through MAPK signaling. Am. J. Physiol. Renal Physiol. 294, F1465F1472 Heilig, C. W., Liu, Y., England, R. L., Freytag, S. O., Gilbert, J. D., Heilig, K. O., Zhu, M., Concepcion, L. A. and Brosius, III, F. C. (1997) D-glucose stimulates mesangial cell GLUT1 expression and basal and IGF-I-sensitive glucose uptake in rat mesangial cells: implications for diabetic nephropathy. Diabetes 46, 10301039 Heilig, C. W., Concepcion, L. A., Riser, B. L., Freytag, S. O., Zhu, M. and Cortes, P (1995) Overexpression of glucose . transporters in rat mesangial cells cultured in a normal glucose milieu mimics the diabetic phenotype. J. Clin. Invest. 96, 18021814

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Weigert, C., Brodbeck, K., Brosius, III, F. C., Huber, M., Lehmann, R., Friess, U., Facchin, S., Aulwurm, S., Haring, H. U., Schleicher, E. D. et al. (2003) Evidence for a novel TGF-1-independent mechanism of bronectin production in mesangial cells overexpressing glucose transporters. Diabetes 52, 527535 Alarcon, C., Morales, A. V., Pimentel, B., Serna, J. and de Pablo, F. (1998) (Pro)insulin and insulin-like growth factor I complementary expression and roles in early development. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 121, 1317 Masters, B. A., Werner, H., Roberts, Jr, C. T., LeRoith, D. and Raizada, M. K. (1991) Developmental regulation of insulin-like growth factor-I-stimulated glucose transporter in rat brain astrocytes. Endocrinology 128, 25482557 Hernandez, E. R., Hurwitz, A., Botero, L., Ricciarelli, E., Werner, H., Roberts, Jr, C. T., LeRoith, D. and Adashi, E. Y. (1991) Insulin-like growth factor receptor gene expression in the rat ovary: divergent regulation of distinct receptor species. Mol. Endocrinol. 5, 17991805 Werner, H., Stannard, B., Bach, M. A., Roberts, Jr, C. T. and LeRoith, D. (1991) Regulation of insulin-like growth factor I receptor gene expression in normal and pathological states. Adv. Exp. Med. Biol. 293, 263272 Kurokawa, K. and Lerner, R. L. (1980) Binding and degradation of insulin by isolated renal cortical tubules. Endocrinology 106, 655662 Pillion, D. J., Haskell, J. F. and Meezan, E. (1988) Distinct receptors for insulin-like growth factor I in rat renal glomeruli and tubules. Am. J. Physiol. 255, E504E512 Meezan, E. and Freychet, P (1980) Specic insulin receptors in . rat renal glomeruli. Renal Physiol. 3, 7278 Baum, M. (1987) Insulin stimulates volume absorption in the rabbit proximal convoluted tubule. J. Clin. Invest. 79, 11041109 Mandon, B., Siga, E., Chabardes, D., Firsov, D., Roinel, N. and De Roufgnac, C. (1993) Insulin stimulates Na + , Cl , Ca2 + and Mg2 + transports in TAL of mouse nephron: cross-potentiation with AVP Am. J. Physiol. 265, F361F369 . Ito, O., Kondo, Y., Takahashi, N., Kudo, K., Igarashi, Y., Omata, K., Imai, Y. and Abe, K. (1994) Insulin stimulates NaCl transport in isolated perfused MTAL of Henles loop of rabbit kidney. Am. J. Physiol. 267, F265F270 Takahashi, N., Ito, O. and Abe, K. (1996) Tubular effects of insulin. Hypertens. Res. 19 (Suppl. 1), S41S45 Fuster, D. G., Bobulescu, I. A., Zhang, J., Wade, J. and Moe, O. W. (2007) Characterization of the regulation of renal Na + /H + exchanger NHE3 by insulin. Am. J. Physiol. Renal Physiol. 292, F577F585 Klisic, J., Hu, M. C., Nief, V., Reyes, L., Fuster, D., Moe, O. W. and Ambuhl, P M. (2002) Insulin activates Na + /H + exchanger . 3: biphasic response and glucocorticoid dependence. Am. J. Physiol. Renal Physiol. 283, F532F539 Sohara, E., Rai, T., Yang, S. S., Ohta, A., Naito, S., Chiga, M., Nomura, N., Lin, S. H., Vandewalle, A., Ohta, E. et al. (2011) Acute insulin stimulation induces phosphorylation of the Na-Cl cotransporter in cultured distal mpkDCT cells and mouse kidney. PLoS ONE 6, e24277 Wang, J., Barbry, P Maiyar, A. C., Rozansky, D. J., Bhargava, A., ., Leong, M., Firestone, G. L. and Pearce, D. (2001) SGK integrates insulin and mineralocorticoid regulation of epithelial sodium transport. Am. J. Physiol. Renal Physiol. 280, F303F313 Zhang, Y. H., Alvarez de la Rosa, D., Canessa, C. M. and Hayslett, J. P (2005) Insulin-induced phosphorylation of ENaC . correlates with increased sodium channel function in A6 cells. Am. J. Physiol. Cell Physiol. 288, C141C147 Biber, J., Murer, H. and Forster, I. (1998) The renal type II Na + /phosphate cotransporter. J. Bioenerg. Biomembr. 30, 187194

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Dai, L. J., Ritchie, G., Bapty, B. W., Kerstan, D. and Quamme, G. A. (1999) Insulin stimulates Mg2 + uptake in mouse distal convoluted tubule cells. Am. J. Physiol. 277, F907F913 Tiwari, S., Riazi, S. and Ecelbarger, C. A. (2007) Insulins impact on renal sodium transport and blood pressure in health, obesity and diabetes. Am. J. Physiol. Renal Physiol. 293, F974F984 Wild, S., Roglic, G., Green, A., Sicree, R. and King, H. (2004) Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes Care 27, 10471053 Ford, E. S., Giles, W. H. and Dietz, W. H. (2002) Prevalence of the metabolic syndrome among US adults: ndings from the third National Health and Nutrition Examination Survey. JAMA, J. Am. Med. Assoc. 287, 356359 Burrows, N. R., Wang, J., Geiss, L. S., Venkat Narayan, K. M. and Engelgau, M. M. (2005) Incidence of end-stage renal disease among persons with diabetes: United States, 19902002. MMWR 54, 10971100 World Health Organization (1999) Denition, Diagnosis and Classication of Diabetes Mellitus and its Complications. Part 1: Diagnosis and Classication of Diabetes Mellitus, World Health Organization, Geneva Schrijvers, B. F., De Vriese, A. S. and Flyvbjerg, A. (2004) From hyperglycemia to diabetic kidney disease: the role of metabolic, hemodynamic, intracellular factors and growth factors/cytokines. Endocr. Rev. 25, 9711010 Pagtalunan, M. E., Miller, P L., Jumping-Eagle, S., Nelson, R. G., . Myers, B. D., Rennke, H. G., Coplon, N. S., Sun, L. and Meyer, T. W. (1997) Podocyte loss and progressive glomerular injury in type II diabetes. J. Clin. Invest. 99, 342348 Haraldsson, B. and Jeansson, M. (2009) Glomerular ltration barrier. Curr. Opin. Nephrol. Hypertens. 18, 331335 Yki-Jarvinen, H. and Koivisto, V. A. (1986) Natural course of insulin resistance in type I diabetes. N. Engl. J. Med. 315, 224230 Andersen, A. R., Christiansen, J. S. andersen, J. K., Kreiner, S. and Deckert, T. (1983) Diabetic nephropathy in Type 1 (insulin-dependent) diabetes: an epidemiological study. Diabetologia 25, 496501 Orchard, T. J., Chang, Y. F., Ferrell, R. E., Petro, N. and Ellis, D. E. (2002) Nephropathy in type 1 diabetes: a manifestation of insulin resistance and multiple genetic susceptibilities? Further evidence from the Pittsburgh Epidemiology of Diabetes Complication Study. Kidney Int. 62, 963970 Hadjadj, S., Pean, F., Gallois, Y., Passa, P Aubert, R., Weekers, ., L., Rigalleau, V., Bauduceau, B., Bekherraz, A., Roussel, R. et al. (2004) Different patterns of insulin resistance in relatives of type 1 diabetic patients with retinopathy or nephropathy: the Genesis France-Belgium Study. Diabetes Care 27, 26612668 Kuboki, K., Jiang, Z. Y., Takahara, N., Ha, S. W., Igarashi, M., Yamauchi, T., Feener, E. P Herbert, T. P Rhodes, C. J. and King, ., ., G. L. (2000) Regulation of endothelial constitutive nitric oxide synthase gene expression in endothelial cells and in vivo: a specic vascular action of insulin. Circulation 101, 676681 Naruse, K., Rask-Madsen, C., Takahara, N., Ha, S. W., Suzuma, K., Way, K. J., Jacobs, J. R., Clermont, A. C., Ueki, K., Ohshiro, Y. et al. (2006) Activation of vascular protein kinase C- inhibits Akt-dependent endothelial nitric oxide synthase function in obesity-associated insulin resistance. Diabetes 55, 691698 Wolf, G., Chen, S. and Ziyadeh, F. N. (2005) From the periphery of the glomerular capillary wall toward the center of disease: podocyte injury comes of age in diabetic nephropathy. Diabetes 54, 16261634 Wang, Y., Heilig, K., Saunders, T., Minto, A., Deb, D. K., Chang, A., Brosius, F., Monteiro, C. and Heilig, C. W. (2010) Transgenic overexpression of GLUT1 in mouse glomeruli produces renal disease resembling diabetic glomerulosclerosis. Am. J. Physiol. Renal Physiol. 299, F99F111

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

Zhang, H., Schin, M., Saha, J., Burke, K., Holzman, L. B., Filipiak, W., Saunders, T., Xiang, M., Heilig, C. W. and Brosius, III, F. C. (2010) Podocyte-specic overexpression of GLUT1 surprisingly reduces mesangial matrix expansion in diabetic nephropathy in mice. Am. J. Physiol. Renal Physiol. 299, F91F98 Jim, B., Ghanta, M., Qipo, A., Fan, Y., Chuang, P Y., Cohen, . H. W., Abadi, M., Thomas, D. B. and He, J. C. (2012) Dysregulated nephrin in diabetic nephropathy of type 2 diabetes: a cross sectional study. PLoS ONE 7, e36041 Patari, A., Forsblom, C., Havana, M., Taipale, H., Groop, P H. . and Holthofer, H. (2003) Nephrinuria in diabetic nephropathy of type 1 diabetes. Diabetes 52, 29692974 Doublier, S., Salvidio, G., Lupia, E., Ruotsalainen, V., Verzola, D., Deferrari, G. and Camussi, G. (2003) Nephrin expression is reduced in human diabetic nephropathy: evidence for a distinct role for glycated albumin and angiotensin II. Diabetes 52, 10231030 Garvey, W. T., Olefsky, J. M. and Marshall, S. (1986) Insulin induces progressive insulin resistance in cultured rat adipocytes. Sequential effects at receptor and multiple postreceptor sites. Diabetes 35, 258267 Venkatesan, N. and Davidson, M. B. (1995) Insulin resistance in rats harboring growth hormone-secreting tumors: decreased receptor number but increased kinase activity in liver. Metab., Clin. Exp. 44, 7584 Clerk, L. H., Vincent, M. A., Barrett, E. J., Lankford, M. F. and Lindner, J. R. (2007) Skeletal muscle capillary responses to insulin are abnormal in late-stage diabetes and are restored by angiotensin-converting enzyme inhibition. Am. J. Physiol. Endocrinol. Metab. 293, E1804E1809 Catena, C., Cavarape, A., Novello, M., Giacchetti, G. and Sechi, L. A. (2003) Insulin receptors and renal sodium handling in hypertensive fructose-fed rats. Kidney Int. 64, 21632171 Catena, C., Giacchetti, G., Novello, M., Colussi, G., Cavarape, A. and Sechi, L. A. (2003) Cellular mechanisms of insulin resistance in rats with fructose-induced hypertension. Am. J. Hypertens. 16, 973978 Mogensen, C. E., Christensen, N. J. and Gundersen, H. J. (1980) The acute effect of insulin on heart rate, blood pressure, plasma noradrenaline and urinary albumin excretion. The role of changes in blood glucose. Diabetologia 18, 453457 Hegedus, L., Christensen, N. J., Mogensen, C. E. and Gundersen, H. J. (1980) Oral glucose increases urinary albumin excretion in normal subjects but not in insulin-dependent diabetics. Scand. J. Clin. Lab. Invest. 40, 479482 Corman, B., Chami-Khazraji, S., Schaeverbeke, J. and Michel, J. B. (1988) Effect of feeding on glomerular ltration rate and proteinuria in conscious aging rats. Am. J. Physiol. 255, F250F256 Uemasu, J., Hori, T., Uemasu, Y. and Kawasaki, H. (1991) Effects of a rice meal on renal hemodynamics and excretory functions in normal subjects. Nephron 57, 187191 Isshiki, K., Haneda, M., Koya, D., Maeda, S., Sugimoto, T. and Kikkawa, R. (2000) Thiazolidinedione compounds ameliorate glomerular dysfunction independent of their insulin-sensitizing action in diabetic rats. Diabetes 49, 10221032 Panchapakesan, U., Chen, X. M. and Pollock, C. A. (2005) Drug insight: thiazolidinediones and diabetic nephropathyrelevance to renoprotection. Nat. Clin. Pract. Nephrol. 1, 3343 Saradis, P A., Stafylas, P C., Georgianos, P I., Saratzis, A. N. . . . and Lasaridis, A. N. (2010) Effect of thiazolidinediones on albuminuria and proteinuria in diabetes: a meta-analysis. Am. J. Kidney Dis. 55, 835847 Ma, L. J., Marcantoni, C., Linton, M. F., Fazio, S. and Fogo, A. B. (2001) Peroxisome proliferator-activated receptor- agonist troglitazone protects against nondiabetic glomerulosclerosis in rats. Kidney Int. 59, 18991910

368

The Authors Journal compilation

2013 Biochemical Society

Insulin and the kidney

172

173

174

175

176

177

178

179 180 181

182

183

184

185

186

187

188

Yang, H. C., Deleuze, S., Zuo, Y., Potthoff, S. A., Ma, L. J. and Fogo, A. B. (2009) The PPAR agonist pioglitazone ameliorates aging-related progressive renal injury. J. Am. Soc. Nephrol. 20, 23802388 Lennon, R., Welsh, G. I., Singh, A., Satchell, S. C., Coward, R. J., Tavare, J. M., Mathieson, P W. and Saleem, M. A. (2009) . Rosiglitazone enhances glucose uptake in glomerular podocytes using the glucose transporter GLUT1. Diabetologia 52, 19441952 Lennon, R., Pons, D., Sabin, M. A., Wei, C., Shield, J. P ., Coward, R. J., Tavare, J. M., Mathieson, P W., Saleem, M. A. . and Welsh, G. I. (2009) Saturated fatty acids induce insulin resistance in human podocytes: implications for diabetic nephropathy. Nephrol. Dial. Transplant. 24, 32883296 Hyvonen, M. E., Saurus, P Wasik, A., Heikkila, E., Havana, M., ., Trokovic, R., Saleem, M., Holthofer, H. and Lehtonen, S. (2010) Lipid phosphatase SHIP2 downregulates insulin signalling in podocytes. Mol. Cell. Endocrinol. 328, 7079 Lee, M. P and Sweeney, G. (2006) Insulin increases gelatinase . activity in rat glomerular mesangial cells via ERK- and PI-3 kinase-dependent signalling. Diabetes Obes. Metab 8, 281288 Isshiki, K., He, Z., Maeno, Y., Ma, R. C., Yasuda, Y., Kuroki, T., White, G. S., Patti, M. E., Weir, G. C. and King, G. L. (2008) Insulin regulates SOCS2 expression and the mitogenic effect of IGF-1 in mesangial cells. Kidney Int. 74, 14341443 Ling, B. N., Seal, E. E. and Eaton, D. C. (1993) Regulation of mesangial cell ion channels by insulin and angiotensin II. Possible role in diabetic glomerular hyperltration. J. Clin. Invest. 92, 21412151 Wullschleger, S., Loewith, R. and Hall, M. N. (2006) TOR signaling in growth and metabolism. Cell 124, 471484 Hay, N. and Sonenberg, N. (2004) Upstream and downstream of mTOR. Genes Dev. 18, 19261945 Huber, T. B., Walz, G. and Kuehn, E. W. (2011) mTOR and rapamycin in the kidney: signaling and therapeutic implications beyond immunosuppression. Kidney Int. 79, 502511 Takano, A., Usui, I., Haruta, T., Kawahara, J., Uno, T., Iwata, M. and Kobayashi, M. (2001) Mammalian target of rapamycin pathway regulates insulin signaling via subcellular redistribution of insulin receptor substrate 1 and integrates nutritional signals and metabolic signals of insulin. Mol. Cell. Biol. 21, 50505062 Lamming, D. W., Ye, L., Katajisto, P Goncalves, M. D., Saitoh, ., M., Stevens, D. M., Davis, J. G., Salmon, A. B., Richardson, A., Ahima, R. S. et al. (2012) Rapamycin-induced insulin resistance is mediated by mTORC2 loss and uncoupled from longevity. Science 335, 16381643 Lieberthal, W. and Levine, J. S. (2009) The role of the mammalian target of rapamycin (mTOR) in renal disease. J. Am. Soc. Nephrol. 20, 24932502 Inoki, K., Mori, H., Wang, J., Suzuki, T., Hong, S., Yoshida, S., Blattner, S. M., Ikenoue, T., Ruegg, M. A., Hall, M. N. et al. (2011) mTORC1 activation in podocytes is a critical step in the development of diabetic nephropathy in mice. J. Clin. Invest. 121, 21812196 Godel, M., Hartleben, B., Herbach, N., Liu, S., Zschiedrich, S., Lu, S., Debreczeni-Mor, A., Lindenmeyer, M. T., Rastaldi, M. P ., Hartleben, G. et al. (2011) Role of mTOR in podocyte function and diabetic nephropathy in humans and mice. J. Clin. Invest. 121, 21972209 Cina, D. P Onay, T., Paltoo, A., Li, C., Maezawa, Y., De Arteaga, ., J., Jurisicova, A. and Quaggin, S. E. (2011) Inhibition of MTOR disrupts autophagic ux in podocytes. J. Am. Soc. Nephrol. 23, 412420 Lloberas, N., Cruzado, J. M., Franquesa, M., Herrero-Fresneda, I., Torras, J., Alperovich, G., Rama, I., Vidal, A. and Grinyo, J. M. (2006) Mammalian target of rapamycin pathway blockade slows progression of diabetic kidney disease in rats. J. Am. Soc. Nephrol. 17, 13951404

189

190

191

192

193

194

195

196

197

198

199

200

201

202

203

204

Yang, Y., Wang, J., Qin, L., Shou, Z., Zhao, J., Wang, H., Chen, Y. and Chen, J. (2007) Rapamycin prevents early steps of the development of diabetic nephropathy in rats. Am. J. Nephrol. 27, 495502 Mori, H., Inoki, K., Masutani, K., Wakabayashi, Y., Komai, K., Nakagawa, R., Guan, K. L. and Yoshimura, A. (2009) The mTOR pathway is highly activated in diabetic nephropathy and rapamycin has a strong therapeutic potential. Biochem. Biophys. Res. Commun. 384, 471475 Torras, J., Herrero-Fresneda, I., Gulias, O., Flaquer, M., Vidal, A., Cruzado, J. M., Lloberas, N., Franquesa, M. and Grinyo, J. M. (2009) Rapamycin has dual opposing effects on proteinuric experimental nephropathies: is it a matter of podocyte damage? Nephrol. Dial. Transplant. 24, 36323640 Amer, H. and Cosio, F. G. (2009) Signicance and management of proteinuria in kidney transplant recipients. J. Am. Soc. Nephrol. 20, 24902492 Letavernier, E. and Legendre, C. (2008) mToR inhibitors-induced proteinuria: mechanisms, signicance and management. Transplant. Rev. 22, 125130 Letavernier, E., Bruneval, P Mandet, C., Duong Van Huyen, J. P ., ., Peraldi, M. N., Helal, I., Noel, L. H. and Legendre, C. (2007) High sirolimus levels may induce focal segmental glomerulosclerosis de novo. Clin. J. Am. Soc. Nephrol. 2, 326333 Tiwari, S., Sharma, N., Gill, P S., Igarashi, P Kahn, C. R., Wade, . ., J. B. and Ecelbarger, C. M. (2008) Impaired sodium excretion and increased blood pressure in mice with targeted deletion of renal epithelial insulin receptor. Proc. Natl. Acad. Sci. U.S.A. 105, 64696474 Shen, S. S., Krishna, B., Chirala, R., Amato, R. J. and Truong, L. D. (2005) Kidney-specic cadherin, a specic marker for the distal portion of the nephron and related renal neoplasms. Mod. Pathol. 18, 933940 Musso, C., Javor, E., Cochran, E., Balow, J. E. and Gorden, P . (2006) Spectrum of renal diseases associated with extreme forms of insulin resistance. Clin. J. Am. Soc. Nephrol. 1, 616622 Arioglu, E., andewelt, A., Diabo, C., Bell, M., Taylor, S. I. and Gorden, P (2002) Clinical course of the syndrome of . autoantibodies to the insulin receptor (type B insulin resistance): a 28-year perspective. Medicine 81, 87100 Tsokos, G. C., Gorden, P Antonovych, T., Wilson, C. B. and ., Balow, J. E. (1985) Lupus nephritis and other autoimmune features in patients with diabetes mellitus due to autoantibody to insulin receptors. Ann. Intern. Med. 102, 176181 Musso, C., Cochran, E., Moran, S. A., Skarulis, M. C., Oral, E. A., Taylor, S. and Gorden, P (2004) Clinical course of genetic . diseases of the insulin receptor (type A and Rabson-Mendenhall syndromes): a 30-year prospective. Medicine 83, 209222 Woodcock, J., Sharfstein, J. M. and Hamburg, M. (2010) Regulatory action on rosiglitazone by the U.S. Food and Drug Administration. N. Engl. J. Med. 363, 14891491 Kestila, M., Lenkkeri, U., Mannikko, M., Lamerdin, J., McCready, P Putaala, H., Ruotsalainen, V., Morita, T., Nissinen, ., M., Herva, R. et al. (1998) Positionally cloned gene for a novel glomerular proteinnephrinis mutated in congenital nephrotic syndrome. Mol. Cell 1, 575582 Donoviel, D. B., Freed, D. D., Vogel, H., Potter, D. G., Hawkins, E., Barrish, J. P Mathur, B. N., Turner, C. A., Geske, R., ., Montgomery, C. A. et al. (2001) Proteinuria and perinatal lethality in mice lacking NEPH1, a novel protein with homology to NEPHRIN. Mol. Cell. Biol. 21, 48294836 Kaplan, J. M., Kim, S. H., North, K. N., Rennke, H., Correia, L. A., Tong, H. Q., Mathis, B. J., Rodriguez-Perez, J. C., Allen, P . G., Beggs, A. H. et al. (2000) Mutations in ACTN4, encoding -actinin-4, cause familial focal segmental glomerulosclerosis. Nat. Genet. 24, 251256

www.clinsci.org

369

L. J. Hale and R. J. M. Coward

205

206

207

Yang, Y., Jeanpierre, C., Dressler, G. R., Lacoste, M., Niaudet, P . and Gubler, M. C. (1999) WT1 and PAX-2 podocyte expression in DenysDrash syndrome and isolated diffuse mesangial sclerosis. Am. J. Pathol. 154, 181192 Dreyer, S. D., Zhou, G., Baldini, A., Winterpacht, A., Zabel, B., Cole, W., Johnson, R. L. and Lee, B. (1998) Mutations in LMX1B cause abnormal skeletal patterning and renal dysplasia in nail patella syndrome. Nat. Genet. 19, 4750 Kim, J. M., Wu, H., Green, G., Winkler, C. A., Kopp, J. B., Miner, J. H., Unanue, E. R. and Shaw, A. S. (2003) CD2-associated protein haploinsufciency is linked to glomerular disease susceptibility. Science 300, 12981300

208

209

210

Boute, N., Gribouval, O., Roselli, S., Benessy, F., Lee, H., Fuchshuber, A., Dahan, K., Gubler, M. C., Niaudet, P and . Antignac, C. (2000) NPHS2, encoding the glomerular protein podocin, is mutated in autosomal recessive steroid-resistant nephrotic syndrome. Nat. Genet. 24, 349354 Brown, E. J., Schlondorff, J. S., Becker, D. J., Tsukaguchi, H., Uscinski, A. L., Higgs, H. N., Henderson, J. M. and Pollak, M. R. (2009) Mutations in the formin gene INF2 cause focal segmental glomerulosclerosis. Nat. Genet. 42, 7276 Inoue, T., Yaoita, E., Kurihara, H., Shimizu, F., Sakai, T., Kobayashi, T., Ohshiro, K., Kawachi, H., Okada, H., Suzuki, H. et al. (2001) FAT is a component of glomerular slit diaphragms. Kidney Int. 59, 10031012

Received 13 July 2012/14 September 2012; accepted 26 September 2012 Published on the Internet 27 November 2012, doi: 10.1042/CS20120378

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