ABSTRACT
<p><b>OBJECTIVE</b>To analyze the clinical features and WT1 gene mutations in patients with steroid-resistant nephrotic syndrome (SRNS) accompanied with genitourinary malformations. The expression of podocyte molecules was also investigated in renal specimen of these WT1 mutated patients.</p><p><b>METHODS</b>From January 2005 to May 2007, 3 cases of SRNS accompanied with genitourinary malformations were involved in this study. The expression of podocyte molecules (nephrin, podocin, alpha-actinin 4, WT1 and CD2AP) in 2 cases was analyzed by immunofluorescence and immunohistochemistry; using PCR to amplify genomic DNA and RT-PCR to amplify WT1 cDNA. GeneScan and GeneScan software were used to quantify the ratio of +KTS/-KTS isoforms.</p><p><b>RESULTS</b>The age of onset of the 3 cases were 6 months, 1 year and 10 years, respectively. The age at diagnosis was 7 months, 9 years and 15 years, respectively. The phenotype of case 1 and case 3 was male accompanied with genitourinary malformations. Case 2 was phenotypic female. Karyotype analysis of the 3 cases revealed 46, XY. Each case was diagnosed as SRNS. Focal segmental glomerulosclerosis (FSGS) was confirmed in 2 cases. Podocyte molecular expression altered in renal tissues of 2 cases. WT1 staining was negative in case 1. WT1 expression in case 2 showed a diffuse nuclear staining with less obvious speckles compared with controls. WT1 IVS 9 + 5 G > A mutation was detected in case 2 and WT1 exon 9 1186 G > A mutation was detected in case 3. No WT1 mutation was detected in case 1.</p><p><b>CONCLUSIONS</b>Karyotype analysis and WT1 genetic analyzing should be performed for all female patients with early onset SRNS and in male patients with SRNS accompanied with genitourinary malformations. The abnormal ratio of +KTS/-KTS isoforms caused by WT1 mutations along with abnormal expression of podocyte molecules were involved in the pathogenesis of proteinuria.</p>
Subject(s)
Adolescent , Child , Female , Humans , Infant , Male , Mutation , Nephrotic Syndrome , Genetics , Phenotype , Podocytes , Steroids , Urogenital Abnormalities , Genetics , WT1 Proteins , GeneticsABSTRACT
OBJECTIVE@#To understand WT1 mutations in patients with steroid resistant nephrotic syndrome (SRNS) accompanied with genitourinary malformations.@*METHODS@#Three cases of SRNS accompanied with genitourinary malformations were enrolled. The expression of podocyte molecules (nephrin, podocin, alpha-actinin-4, WT1, and CD2AP) in 2 cases was analyzed with the immunofluorescence and immunohistochemistry techniques. The genomic DNA and cDNA of WT1 were analyzed by using PCR and RT-PCR, respectively. GeneScan and GeneScan software were used to quantify the ratio of +KTS/-KTS isoforms.@*RESULTS@#The onset ages of 3 cases were 6 months, 1 year, and 10 years old, respectively. The diagnosis age was 7 months, 9 years, and 15 years old, respectively. The phenotype of Case 1 and Case 3 was male accompanied with genitourinary malformations. Case 2 was phenotypic female. Karyotype analysis of 3 cases revealed 46, XY. Three cases were diagnosed as SRNS. Focal segmental glomerulosclerosis (FSGS) was confirmed in 2 cases. Podocyte molecular expression altered in renal tissues of 2 cases. In addition, WT1 staining was negative in Case 1. WT1 expression in Case 2 showed diffuse nuclear staining with less obvious speckles compared with controls. WT1 IVS 9 +5 G>A mutation was detected in Case 2 and WT1 Exon 9 1186 G>A mutation was detected in Case 3. No WT1 mutation was detected in Case 1.@*CONCLUSION@#Karyotype analysis and WT1 genetic testing should be done in all female patients with early onset steroid resistant FSGS and in male patients with SRNS accompanied with genitourinary malformations. Abnormal podocyte molecular expression suggests that more podocyte molecules might be involved in the pathogenesis of proteinuria in WT1 mutational patients.
Subject(s)
Adolescent , Child , Female , Humans , Infant , Male , Drug Resistance , Genetics , Exons , Hormones , Pharmacology , Mutation , Nephrotic Syndrome , Genetics , Phenotype , Urogenital Abnormalities , Genetics , WT1 Proteins , GeneticsABSTRACT
0.05) at any observed time points as compared with the controls.3.From day 7 after the adriamycin injection,VEGF protein reduced significantly(P
ABSTRACT
Objective To explore the expression and distribution of transient receptor potential cation channel 6(TRPC6)in normal human,mice,rats' renal tissue and the mouse podocyte clone 5(MPC5)for further investigating the relationship between TRPC6 and the protei-nuria-related podocyte molecules.Methods 1.The distributions of TRPC6 in normal human,mice,rats' renal tissue and MPC5 were observed by using the immunochemistry staining.2.The mRNA expression of TRPC6 in mouse renal cortex and differentiated MPC5 was detected by using reverse transcriptase-polymerase chain reaction(RT-PCR).3.The protein expression of TRPC6 in human,mice and differentiated MPC5 was detected by using Western blotting.Results 1.In human kidney,TRPC6 showed a weak staining in glomeruli and a strong staining in renal tubules and vessels.In mice and rats' kidney,TRPC6 showed a strong staining in glomeruli and was mainly distributed along the capillary loops of glomerulus and in mesangium.The positive staining of TRPC6 was observed in MPC5,which was distributed evenly on the cell membrane in differentiated podocytes.2.The specific PCR band of TRPC6 was detected in mouse renal cortex and differentiated MPC5.3.The specific protein band of TRPC6 was detected in normal human,mice renal cortex and differentiated MPC5 with the size of 106.Conclusions The expression of TRPC6 is verified in normal human,mice and rats' kidneys,and in differentiated MPC5.These results will benefit for further exploring the relationship between TRPC6 and the proteinuria-related podocyte molecules.