RESUMO
<p><b>OBJECTIVE</b>To illuminate pathogenic gene and mutation in a Chinese family with autosomal dominant retinitis pigmentosa (adRP).</p><p><b>METHODS</b>Genetic linkage analysis was performed on the known genetic loci for adRP with a panel of polymorphic markers, and then all exons including exon-intron boundary, 5oUTR and 3oUTR of the candidate gene were sequenced directly.</p><p><b>RESULTS</b>Two-point LOD scores were negative with all markers tested except D17S701 (Zmax=2.107, theta=0) and D17S1604 (Zmax=1.806, theta=0). The disease gene locus was confined to RP17 with further genetic linkage and haplotype analysis. Screening all exons including exon-intron boundary, 5oUTR and 3oUTR of carbonic anhydrase 4 (CA4) revealed no mutation in this family.</p><p><b>CONCLUSION</b>The disease-causing gene of one Chinese family with adRP was first mapped to RP17, however no gene mutation of CA4 was detected in this family. Maybe there is a complex CA4 gene mutation in this family or a new disease-causing gene for this family in this locus, further study need to be done.</p>
Assuntos
Feminino , Humanos , Masculino , Povo Asiático , Genética , Anidrase Carbônica IV , Genética , Éxons , Genética , Ligação Genética , Genética , Marcadores Genéticos , Genética , Haplótipos , Genética , Íntrons , Genética , Mutação , Retinose Pigmentar , GenéticaRESUMO
<p><b>OBJECTIVE</b>To demonstrate the molecular expression of carbonic anhydrase IV (CA IV) in rabbit corneal endothelium.</p><p><b>METHODS</b>Reverse transcriptase polymerase chain reaction (RT-PCR) was performed using cultured and fresh rabbit corneal endothelial total RNA and specific primers for CA IV. The RT-PCR product was subcloned and sequenced. Immunoblotting and indirect immunofluorescence staining were performed to detect protein expression and distribution of CA IV using fresh and cultured rabbit corneal endothelium and rat anti-CA IV polyclonal antibody.</p><p><b>RESULTS</b>RT-PCR screening gave positive bands at the predicted size for CA IV from fresh and cultured rabbit corneal endothelium. Sequencing further confirmed the identity of CA IV in corneal endothelium. Immunoblotting analysis showed a single band at 52 kDa for freshly isolated and cultured endothelial cells. Indirect immunofluorescence staining revealed an apparent positive staining in cultured endothelial cells.</p><p><b>CONCLUSION</b>Carbonic anhydrase IV is expressed in rabbit corneal endothelium, which could contribute to the transendothelial HCO(3)(-) flux that is necessary to maintain corneal hydration and transparency.</p>