Rodolfo S. Martin

Mutations in SEC63 cause autosomal dominant polycystic liver disease

Mutations in PRKCSH, encoding the β-subunit of glucosidase II, an N-linked glycan-processing enzyme in the endoplasmic reticulum (ER), cause autosomal dominant polycystic liver disease. We found that mutations in SEC63, encoding a component of the protein translocation machinery in the ER, also cause this disease. These findings are suggestive of a role for cotranslational protein-processing pathways in maintaining epithelial luminal structure and implicate noncilial ER proteins in human polycystic disease.

Identification of a Locus for Autosomal Dominant Polycystic Liver Disease, on Chromosome 19p13.2-13.1

Polycystic liver disease (PCLD) is characterized by the growth of fluid-filled cysts of biliary epithelial origin in the liver. Although the disease is often asymptomatic, it can, when severe, lead to complications requiring surgical therapy. PCLD is most often associated with autosomal dominant polycystic kidney disease (ADPKD); however, families with an isolated polycystic liver phenotype without kidney involvement have been described. The clinical presentation and histological features of polycystic liver disease in the presence or absence of ADPKD are indistinguishable, raising the possibility that the pathogenetic mechanisms in the diseases are interrelated. We ascertained two large families with polycystic liver disease without kidney cysts and performed a genomewide scan for genetic linkage. A causative gene, PCLD, was mapped to chromosome 19p13.2-13.1, with a maximum LOD score of 10.3. Haplotype analysis refined the PCLD interval to 12.5 cM flanked by D19S586/D19S583 and D19S593/D19S579. The discovery of genetic linkage will facilitate diagnosis and study of this underdiagnosed disease entity. Identification of PCLD will be instrumental to an understanding of the pathogenesis of cyst formation in the liver in isolated PCLD and in ADPKD.

Increased Secretion of Potassium in the Rectum of Humans With Chronic Renal Failure

Previous studies have shown that the intestinal excretion of potassium increases in patients with renal failure and serves to guard against severe potassium retention. It is not known, however, whether the rise in intestinal potassium excretion occurs because of an increase in intestinal potassium secretion or a reduction in potassium absorption. Therefore, studies were performed to evaluate the rate of potassium secretion in the human rectum of controls and subjects with renal insufficiency, using a dialysis bag technique. The results demonstrate that net potassium secretion was increased in subjects with renal failure (-5.2 +/- 0.9 microEq X min-1) compared with the control value of -2.0 +/- 0.4 microEq (P less than .05). This change in intestinal secretion of potassium was shown to be independent of the passive effects of plasma potassium. The rise in potassium secretion, however, correlated directly with an increase in transepithelial potential difference (lumen-negative). Although plasma aldosterone levels were higher in patients than in controls, the scatter of data precludes an assessment of the role of aldosterone in the mechanism of the rise in potassium secretion. These data suggest that augmented intestinal potassium excretion in patients with chronic renal insufficiency is caused by increased net potassium secretion in the large intestine, and highlight the role of the intestine in maintaining potassium balance.

Isolated polycystic liver disease not linked to polycystic kidney disease 1 and 2

Autosomal dominant polycystic liver diseaseoccurs commonly in association with autosomal dominantpolycystic kidney disease, types 1 and 2. It may alsoexist as a separate entity, genetically distinct from autosomal dominant polycystic kidneydisease types 1 and 2, as has been recently establishedto exist in a Belgian family. We report here a largeArgentinian family of Spanish-Belgian ancestry with autosomal dominant polycystic liver disease,where proximal and distal markers for both polycystickidney disease 1 and 2 failed to demonstrate geneticlinkage. The data support the notion that polycystic liver disease and autosomal dominant polycystickidney disease may have separate chromosomalloci.

Characteristics of Intracranial Aneurysms in the Else Kröner-Fresenius Registry of Autosomal Dominant Polycystic Kidney Disease

Background: Patients who harbor intracranial aneurysms (IAs) run a risk for aneurysm rupture and subsequent subarachnoid hemorrhage which frequently results in permanent deficits or death. Prophylactic treatment of unruptured aneurysms is possible and recommended depending on the size and location of the aneurysm as well as patient age and condition. IAs are major manifestations of autosomal dominant polycystic kidney disease (ADPKD). Current guidelines do not suggest surveillance of IAs in ADPKD except in the setting of family history if IA was known in any relative with ADPKD. Management of IAs in ADPKD is problematic because limited data exist from large studies. Methods: We established the Else Kröner-Fresenius Registry for ADPKD in Germany. Clinical data were assessed for age at diagnosis of IAs, stage of renal insufficiency, and number, location and size of IAs as well as family history of cerebral events. Patients with symptomatic or asymptomatic IAs were included. All patients with ADPKD-related IAs were offered mutation scanning of the susceptibility genes for ADPKD, the PKD1 and PKD2 genes. Results: Of 463 eligible ADPKD patients from the population base of Germany, 32 (7%) were found to have IAs, diagnosed at the age of 2–71 years, 19 females and 13 males. Twenty (63%) of these 32 patients were symptomatic, whereas IAs were detected in an asymptomatic stage in 12 patients. IAs were multifocal in 12 and unifocal in 20 patients. In 26 patients (81%), IAs were diagnosed before end-stage renal failure. Twenty-five out of 27 unrelated index cases (93%) had no IAs or cerebral events documented in their relatives with ADPKD. In 16 unrelated index patients and 3 relatives, we detected germline mutations. The mutations were randomly distributed across the PKD1 gene in 14 and the PKD2 gene in 2 index cases. Questionnaires answered for 320/441 ADPKD patients without IAs revealed that only 45/320 (14%) had MR angiography. Conclusion: In ADPKD, rupture of IAs occurs frequently before the start of dialysis, is only infrequently associated with a family history of IAs or subarachnoid hemorrhage, and is associated with mutations either of the PKD1 or the PKD2 gene of any type. Screening for IAs is widely insufficiently performed, should not be restricted to families with a history of cerebral events and should be started before end-stage renal failure.

Early renal and vascular changes in ADPKD patients with low-grade albumin excretion and normal renal function

BACKGROUND Autosomal dominant polycystic kidney disease (ADPKD) shows an increase in both urine monocyte chemoattractant protein-1 (MCP-1) and carotid intima-media thickness (CIMT) before changes in serum creatinine concentration. Although microalbuminuria is an index of disease progression, data on whether renal alterations and vascular remodelling are already present at normal or minimally increased levels of urine albumin excretion in early stages of the disease are lacking. METHODS Forty-eight ADPKD patients (24.8 +/- 0.8 years) with normal renal function (MDRD 108.1 +/- 3.1 ml/min) and 21 age-matched controls were studied in a cross-sectional study. The urine albumin/creatinine ratio (UACR) above the upper range of controls (6.8 mg/g) was taken as the predictor of renal alterations and vascular remodelling. Urine MCP-1, MCP-1 fractional excretion (FE(MCP-1)), endothelial-dependent vascular relaxation (EDVR), aortic pulse-wave velocity (Ao-PWV) and CIMT were chosen as biological markers. RESULTS No differences between ADPKD with UACR <or=6.8 mg/g and controls were observed in urine MCP-1 (77.7 +/- 13.9 versus 57.8 +/- 6.3 ng/g), FE(MCP-1) (91 +/- 19 versus 74 +/- 8%) and CIMT (0.47 +/- 0.06 versus 0.44 +/- 0.07 mm), respectively. Conversely, ADPKD with UACR >6.8 mg/g showed values that were different from the two other groups. In addition, patients with UACR >6.8 and <20 mg/g showed greater values for urine MCP-1, FE(MCP-1) and CIMT (131.8 +/- 21.7 ng/g, 159 +/- 31% and 0.55 +/- 0.05 mm, respectively), as compared with patients with UACR <or=6.8 mg/g. The same pattern was found in a subset of normotensive ADPKD patients. No differences were found in EDVR and Ao-PWV. CONCLUSION In young ADPKD patients, normal levels of UACR suggest that renal interstitium is comparable to that in healthy subjects and indicate an absence of subtle atherosclerotic changes in the carotid arteries. Likewise, early renal and vascular changes may be present at UACR below the levels defined as microalbuminuria.

Genetic heterogeneity of autosomal dominant polycystic kidney disease in Argentina.

Autosomal dominant polycystic kidney disease (ADPKD) is an inherited disorder with genetic heterogeneity. Up to three loci are involved in this disease, PKD1 on chromosome 16p13.3, PKD2 on 4q21, and a third locus of unknown location. Here we report the existence of locus heterogeneity for this disease in the Argentinian population by performing linkage analysis on 12 families of Caucasian origin. Eleven families showed linkage to PKD 1 and one family showed linkage to PKD2. Two recombinants in the latter family placed the locus PKD2 proximal to D4S1563, in agreement with data recently published on the cloning of this gene. Analysis of clinical data suggests a milder ADPKD phenotype for the PKD2 family.

Role of aldosterone in the mechanism of renal potassium adaptation

Chronic potassium loading results in an adaptive change in renal tubular epithelium which increases the capacity for potassium excretion. The present study was performed to evaluate the role of aldosterone in renal potassium adaptation, since hyperaldosteronism stimulates potassium secretion, and potassium loading increases the production of aldosterone. Experiments were performed in animals with intact adrenal glands, and in adrenalectomized animals (Adx) replaced with basal physiologic amounts of corticosterone, which 1) were not replaced with aldosterone, or 2) were chronically infused with aldosterone to achieve either basal plasma levels or elevated levels.Chronic potassium loading in adrenal intact animals was associated with a statistically significant higher rate of urinary potassium excretion (3.57±0.30 μEq/min/100 g BW) compared to the control rate (2.54±0.25 μEq/min/100 g BW, p<0.05), during acute infusion of KCl. In potassium loaded Adx animals, with selective replacement of adrenal hormones, the maximum rate of potassium excretion was blunted in the absence of aldosterone, compared to potassium loaded animals with intact adrenal glands. In contrast, when Adx animals were infused chronically with aldosterone, to achieve basal or elevated plasma levels, the maximum rate of potassium excretion was not blunted, although at basal aldosterone levels increased potassium excretion was due, at least in part, to hyperkalemia.These results indicate that the infuction of renal potassium adaptation after a week or more of chronic potassium loading is dependent on the action of hyperaldosteronism on renal tubular epithelium.

Gonadectomy influences blood pressure through the kallikrein-kinin system.

The kallikrein-kinin system (KKS) appears to be involved in blood pressure regulation. We showed that ovariectomy (oVx) stimulates urinary kallikrein activity (UKa). So, we test whether gonadectomy (Gx) would affect blood pressure through an increase in KKS activity and which mechanism(s) were involved. We studied adult Wistar rats of either sex, with and without Gx. At baseline all groups were normotensive although the oVx mean arterial pressure (MAP) was lower than female MAP (p < 0.05). KKS blockade by aprotinin increased MAP (p < 0.05) exclusively in the oVx group. The probably mechanism(s) involved in KKS regulation (synthesis, renal content and UKa) were also studied. Previous Gx, kallikrein content (nkat/g kidney weight) and UKa (nkat/g kidney weight/day) were higher in female than in male rats: 12 ± 1.1 versus 6 ± 0.7 and 40 ± 6.8 versus 26 ± 3.4, respectively. After Gx, kallikrein content increased significantly in both orchiectomized (oRx) and oVx rats, and UKa showed a similar tendency (NS). Kallikrein synthesis did not show gender difference in non-Gx rats, but an increase after oVx was observed. KKS was found to be involved in blood pressure regulation in oVx animals. oVx may trigger the increase in kallikrein synthesis and content and UKa to act upon blood pressure.

Mechanisms of PKC-Dependent Na+ K+ ATPase Phosphorylation in the Rat Kidney with Chronic Renal Failure

The present work was designed to study Na+ K+ ATPase α1-subunit phosphorylation in rats with chronic renal failure (CRF) in comparison with normal rats. Na+ K+ ATPase α1-subunit phosphorylation degree was measured by binding the McK-1 antibody to dephosphorylated Ser-23 in microdissected medullary thick ascending limb of Henle (mTAL) segments. In addition, the total Na+ K+ ATPase α1-subunit expression and activity were also measured in the outer renal medulla homogenates and membranes. CRF rats showed a higher Na+ K+ ATPase activity, as compared with control rats (18.95 ± 2.4 vs. 11.21 ± 1.5 μmol Pi/mg prot/h, p < 0.05), accompanied by a higher total Na+ K+ ATPase expression (0.54 ± 0.04 vs. 0.27 ± 0.02 normalized arbitrary units (NU), p < 0.05). When McK-1 antibody was used, a higher immunosignal in mTAL of CRF rats was observed, as compared with controls (6.3 ± 0.35 vs.4.1 ± 0.33 NU, p < 0.05). The ratio Na+ K+ ATPase α1-subunit phosphorylation / total Na+ K+ ATPase α1-subunit expression per μg protein showed a non-significant difference between CRF and control rats in microdissected mTAL segments (2.11 ± 0.12 vs.2.26 ± 0.18 NU, p = NS). The PKC inhibitor RO-318220 10−6M increased immunosignal (lower phosphorylation degree) in mTAL of CRF rats to 128.43 ± 7.08% (p < 0.05) but did not alter McK1 binding in control rats. Both phorbol 12-myristate 13-acetate (PMA) 10−6M and dopamine 10−6M decreased immunosignal in CRF rats, corresponding to a higher Na+ K+ ATPase α1-subunit phosphorylation degree at Ser-23 (55.26 ± 11.17% and 53.27 ± 7.12% compared with basal, p < 0.05). In mTAL of CRF rats, the calcineurin inhibitor FK-506 10−6M did not modify phosphorylation degree at Ser-23 of Na+ K+ ATPase α1-subunit (100.21 ± 3.00% compared with basal CRF). In control rats, FK 506 10−6M decreased the immunosignal, which corresponds to a higher Na+ K+ ATPase α1-subunit phosphorylation degree at Ser-23. The data suggest that the regulation of basal Na+ K+ ATPase α1-subunit phosphorylation degree at Ser-23 in mTAL segments of CRF rats was primarily dependent on PKC activation rather than calcineurin dependent mechanisms.