title: Alternative splicing events is not a key event for gene expression regulation in uremia.
abstract: BACKGROUND
  The control of gene expression in the course of chronic kidney disease (CKD) is not well addressed. Alternative splicing is a common way to increase complexity of proteins. More than 90% of human transcripts are alternatively spliced. We hypothesised that CKD can induce modification of the alternative splicing machinery.
  
  
  METHODS
  During mutation screening in autosomal dominant polycystic kidney disease, we identified in mononuclear cells (PBMC), an alternative splicing event on the exon 30 of PKD1 gene, the gene implicated in this disease. This alternative splice variant was not correlated with the cystic disease but with CKD. To confirm the association between this variant and CKD, a monocentric clinical study was performed with 3 different groups according to their kidney function (CKD5D, CKD3-5 and normal kidney function). An exon microarray approach was used to highlight splicing events in whole human genome in a normal cell model (fibroblasts) incubated with uremic serum. Alternative splicing variants identified were confirmed by RT-PCR.
  
  
  RESULTS
  The splicing variant of the exon 30 of PKD1 was more frequent in PBMCs from patients with CKD compared to control. With the microarray approach, despite the analysis of more than 230 000 probes, we identified 36 genes with an abnormal splicing index evocating splicing event in fibroblasts exposed to uremic serum. Only one abnormal splicing event in one gene, ADH1B, was confirmed by RT-PCR.
  
  
  CONCLUSIONS
  We observed two alternative spliced genes in two different cell types associated with CKD. Alternative splicing could play a role in the control of gene expression during CKD but it does not seem to be a major mechanism.
authors.affiliation: Aix-Marseille Université, INSERM UMR_S 1076, UFR Pharmacie, Marseille, France ; Centre de Néphrologie et Transplantation Rénale, Assistance Publique-Hôpitaux de Marseille, Hôpital de La Conception, Marseille, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Sallée
authors.affiliation: Aix-Marseille Université, INSERM UMR_S 1062 UFR Médecine, Marseille, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Fontès
authors.affiliation: Aix-Marseille Université, Plate-forme génomique et transcriptomique, UMR_S 910, UFR médecine, Marseille, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Louis
authors.affiliation: Aix-Marseille Université, INSERM UMR_S 1076, UFR Pharmacie, Marseille, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Cérini
authors.affiliation: Aix-Marseille Université, INSERM UMR_S 1076, UFR Pharmacie, Marseille, France ; Centre de Néphrologie et Transplantation Rénale, Assistance Publique-Hôpitaux de Marseille, Hôpital de La Conception, Marseille, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Brunet
authors.affiliation: Aix-Marseille Université, INSERM UMR_S 1076, UFR Pharmacie, Marseille, France ; Centre de Néphrologie et Transplantation Rénale, Assistance Publique-Hôpitaux de Marseille, Hôpital de La Conception, Marseille, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Burtey
mesh_terms: D000368:Aged; D017398:Alternative Splicing / Q000502:physiology*; D016022:Case-Control Studies; D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D046228:Microarray Analysis; D008875:Middle Aged; D051436:Renal Insufficiency, Chronic / Q000235:genetics; D050396:TRPP Cation Channels / Q000235:genetics; D014511:Uremia / Q000235:genetics*
chemical_list: D050396:TRPP Cation Channels