Roland Schmitt

Developmental expression of sodium entry pathways in rat nephron

During the past several years, sites of expression of ion transport proteins in tubules from adult kidneys have been described and correlated with functional properties. Less information is available concerning sites of expression during tubule morphogenesis, although such expression patterns may be crucial to renal development. In the current studies, patterns of renal axial differentiation were defined by mapping the expression of sodium transport pathways during nephrogenesis in the rat. Combined in situ hybridization and immunohistochemistry were used to localize the Na-Pi cotransporter type 2 (NaPi2), the bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2), the thiazide-sensitive Na-Cl cotransporter (NCC), the Na/Ca exchanger (NaCa), the epithelial sodium channel (rENaC), and 11β-hydroxysteroid dehydrogenase (11HSD). The onset of expression of these proteins began in post-S-shape stages. NKCC2 was initially expressed at the macula densa region and later extended into the nascent ascending limb of the loop of Henle (TAL), whereas differentiation of the proximal tubular part of the loop of Henle showed a comparatively retarded onset when probed for NaPi2. The NCC was initially found at the distal end of the nascent distal convoluted tubule (DCT) and later extended toward the junction with the TAL. After a period of changing proportions, subsegmentation of the DCT into a proximal part expressing NCC alone and a distal part expressing NCC together with NaCa was evident. Strong coexpression of rENaC and 11HSD was observed in early nascent connecting tubule (CNT) and collecting ducts and later also in the distal portion of the DCT. Ontogeny of the expression of NCC, NaCa, 11HSD, and rENaC in the late distal convolutions indicates a heterogenous origin of the CNT. These data present a detailed analysis of the relations between the anatomic differentiation of the developing renal tubule and the expression of tubular transport proteins.During the past several years, sites of expression of ion transport proteins in tubules from adult kidneys have been described and correlated with functional properties. Less information is available concerning sites of expression during tubule morphogenesis, although such expression patterns may be crucial to renal development. In the current studies, patterns of renal axial differentiation were defined by mapping the expression of sodium transport pathways during nephrogenesis in the rat. Combined in situ hybridization and immunohistochemistry were used to localize the Na-Pi cotransporter type 2 (NaPi2), the bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2), the thiazide-sensitive Na-Cl cotransporter (NCC), the Na/Ca exchanger (NaCa), the epithelial sodium channel (rENaC), and 11beta-hydroxysteroid dehydrogenase (11HSD). The onset of expression of these proteins began in post-S-shape stages. NKCC2 was initially expressed at the macula densa region and later extended into the nascent ascending limb of the loop of Henle (TAL), whereas differentiation of the proximal tubular part of the loop of Henle showed a comparatively retarded onset when probed for NaPi2. The NCC was initially found at the distal end of the nascent distal convoluted tubule (DCT) and later extended toward the junction with the TAL. After a period of changing proportions, subsegmentation of the DCT into a proximal part expressing NCC alone and a distal part expressing NCC together with NaCa was evident. Strong coexpression of rENaC and 11HSD was observed in early nascent connecting tubule (CNT) and collecting ducts and later also in the distal portion of the DCT. Ontogeny of the expression of NCC, NaCa, 11HSD, and rENaC in the late distal convolutions indicates a heterogenous origin of the CNT. These data present a detailed analysis of the relations between the anatomic differentiation of the developing renal tubule and the expression of tubular transport proteins.

Identification of a Novel A-kinase Anchoring Protein 18 Isoform and Evidence for Its Role in the Vasopressin-induced Aquaporin-2 Shuttle in Renal Principal Cells

Arginine vasopressin (AVP) increases the water permeability of renal collecting duct principal cells by inducing the fusion of vesicles containing the water channel aquaporin-2 (AQP2) with the plasma membrane (AQP2 shuttle). This event is initiated by activation of vasopressin V2 receptors, followed by an elevation of cAMP and the activation of protein kinase A (PKA). The tethering of PKA to subcellular compartments by protein kinase A anchoring proteins (AKAPs) is a prerequisite for the AQP2 shuttle. During the search for AKAP(s) involved in the shuttle, a new splice variant of AKAP18, AKAP18δ, was identified. AKAP18δ functions as an AKAP in vitro and in vivo. In the kidney, it is mainly expressed in principal cells of the inner medullary collecting duct, closely resembling the distribution of AQP2. It is present in both the soluble and particulate fractions derived from renal inner medullary tissue. Within the particulate fraction, AKAP18δ was identified on the same intracellular vesicles as AQP2 and PKA. AVP not only recruited AQP2, but also AKAP18δ to the plasma membrane. The elevation of cAMP caused the dissociation of AKAP18δ and PKA. The data suggest that AKAP18δ is involved in the AQP2 shuttle.

The impact of aging on kidney repair

The process of normal aging affects organ homeostasis as well as responses to acute and chronic injury. In view of the rapid growth in the elderly population, it is increasingly important for us to develop a mechanistic understanding of how these age-dependent changes can impact the susceptibility and response of the kidney to injurious stimuli. In this overview, we focus on the current understanding of those mechanisms by reviewing how cellular changes in the aging kidney might lead to a diminished proliferative reserve, an increased tendency for apoptosis, alterations in growth factor profiles, and changes in potential progenitor and immune cell functions. A better understanding of these processes may help us to define new targets for studying kidney repair and could ultimately lead to new therapeutic strategies that are specifically tailored for treatment of the elderly population.

Protective Effects of B2 Preservation Solution in Comparison to a Standard Solution (Histidine-Tryptophan-Ketoglutarate/Bretschneider) in a Model of Isolated Autologous Hemoperfused Porcine Kidney

Reperfusion injuries after organ transplantation affect graft function and influence long-term graft survival. As hypothermic storage, which minimizes the extent of unspecific tissue injury after ischemia and reperfusion, is significantly influenced by the composition of preservation solutions, strategies to optimize the different components may lead to longer graft survival. In the present study the effects of the preservation solution B2 on early renal function and histopathological changes were compared to histidine-tryptophan-ketoglutarate solution (HTK, Bretschneider) in a model of isolated blood-perfused porcine kidneys. B2-preserved kidneys displayed a lower renal resistance and significantly better creatinine clearance as compared to HTK. Mean differences were also found for filtration fraction and sodium fraction reabsorption. The functional data were also related to histopathological changes. Together, these data indicate that the recently developed preservation solution B2 offers new principles of preservation and is a useful preservation solution for experimental isolated perfused kidney models. B2 may also be an interesting model for optimizing preservation within other organ perfusion models.

Zinc-α2-Glycoprotein Exerts Antifibrotic Effects in Kidney and Heart

Zinc-α2-glycoprotein (AZGP1) is a secreted protein synthesized by epithelial cells and adipocytes that has roles in lipid metabolism, cell cycling, and cancer progression. Our previous findings in AKI indicated a new role for AZGP1 in the regulation of fibrosis, which is a unifying feature of CKD. Using two models of chronic kidney injury, we now show that mice with genetic AZGP1 deletion develop significantly more kidney fibrosis. This destructive phenotype was rescued by injection of recombinant AZGP1. Exposure of AZGP1-deficient mice to cardiac stress by thoracic aortic constriction revealed that antifibrotic effects were not restricted to the kidney but were cardioprotective. In vitro, recombinant AZGP1 inhibited kidney epithelial dedifferentiation and antagonized fibroblast activation by negatively regulating TGF-β signaling. Patient sera with high levels of AZGP1 similarly attenuated TGF-β signaling in fibroblasts. Taken together, these findings indicate a novel role for AZGP1 as a negative regulator of fibrosis progression, suggesting that recombinant AZGP1 may have translational effect for treating fibrotic disease.

Hypothyroidism induces expression of the peptide transporter PEPT2

Abstract The kidney is a target organ for thyroid hormone action and a variety of renal transport processes are altered in response to impaired thyroid functions. To investigate the effect of thyroid hormone on the expression of the renal proximal tubular high-affinity-type H+-peptide cotransporter (PEPT2) in rats, hypothyroidism was induced in animals by administration of methimazole (0.05%) via drinking water. After 7 weeks of treatment, hypothyroidism was confirmed by determining serum free T3 and free T4 concentrations. Northern blotting was used to examine the expression of PEPT2 mRNA in kidney tissues from hypothyroid rats compared to control rats. Hypothyroidism resulted in an increased level of total renal PEPT2 mRNA (121.1±3.3% vs. control 100±2.8%; p=0.008). The mRNA results were confirmed by immuno-blotting, which demonstrated significantly increased protein levels (162% vs. control 100%; p<0.01). Immunohistochemistry also revealed increased PEPT2 protein levels in the proximal tubules of treated compared to non-treated rats. In summary, PEPT2 is the first proximal tubule transporter protein that shows increased expression in states of hypothyreosis. As PEPT2 reabsorbs filtered di- and tripeptides and peptide-like drugs, the present findings may have important implications in nutritional amino acid homeostasis and for drug dynamics in states of altered thyroid function.

Letter to the EditorHDL in CKD: not protective any longer?

To the Editor: With great interest we read the article ‘LDL cholesterol in CKD—to treat or not to treat?,’ in which Massy and Zeeuw reviewed the clinical impact of dyslipidemia on cardiovascular disease in chronic kidney disease (CKD).1 The authors suggest a practical treatment strategy mainly considering study results from the 4D, AURORA, and SHARP studies. Although the focus of the review is on low-density lipoprotein (LDL), we believe that the role of uremic high-density lipoprotein (HDL) should also be considered more carefully in this context. New data clearly support the concept that CKD modulates the level, composition, and functionality of HDL. It was shown that HDL isolated from hemodialysis patients is less effective than normal HDL in accepting cholesterol and causes a significant inflammatory response by inducing interleukin-1 β, interleukin-6, and tumor necrosis factor-α.2 The lipidome of uremic HDL contained increased levels of triglyceride and lysophospholipid, but reduced phospholipids.3 Most recently, Speer et al. described that symmetric dimethylarginine in CKD patients can trigger the transformation from physiological HDL into abnormal HDL, inducing endothelial dysfunction, hypertension, and inflammation through recognition by Toll-like receptor 2.4 As these data argue for a reversed effect of otherwise beneficial HDL in CKD patients, we suggest that altered HDL levels should always be considered in conjunction with LDL when evaluating the lipid status and planning treatment strategies in CKD patients.

Potential Impact of Dialysate Magnesium on Intradialytic Hypotension

Numerous beneficial effects on cardiovascular health have been described for magnesium (Mg). Intradialytic hypotension (IDH) is a common complication in hemodialysis patients which contributes to cardiovascular mortality. It has been suggested that higher dialysate Mg (DMg) might reduce the risk of IDH.

ZAG—a novel biomarker for cardiovascular risk in ESRD patients?

End-stage renal disease (ESRD) is associated with an exceedingly high risk of cardiovascular disease. Traditional risk factors function differently in ESRD, which has prompted a search for novel mechanisms and biomarkers. In an observational study, Bouchara etxa0al. identified zinc-alpha2-glycoprotein (ZAG) as a prognostic marker for cardiovascular events and mortality in patients on dialysis. Although the study raises important questions, the results should be interpreted cautiously and need to be confirmed in independent studies.