Asunto(s)
Fármacos Anti-VIH/uso terapéutico , Infecciones por VIH/tratamiento farmacológico , VIH-1/efectos de los fármacos , Fármacos Anti-VIH/efectos adversos , Fármacos Anti-VIH/clasificación , Antagonistas de los Receptores CCR5 , Ciclohexanos/efectos adversos , Ciclohexanos/uso terapéutico , Inhibidores de Fusión de VIH/efectos adversos , Inhibidores de Fusión de VIH/uso terapéutico , Infecciones por VIH/virología , Inhibidores de Integrasa VIH/efectos adversos , Inhibidores de Integrasa VIH/uso terapéutico , Humanos , Maraviroc , Piperazinas/efectos adversos , Piperazinas/uso terapéutico , Pirimidinas/efectos adversos , Pirimidinas/uso terapéutico , Pirrolidinonas/efectos adversos , Pirrolidinonas/uso terapéutico , Quinolonas/efectos adversos , Quinolonas/uso terapéutico , Raltegravir Potásico , Succinatos/efectos adversos , Succinatos/uso terapéutico , Triazoles/efectos adversos , Triazoles/uso terapéutico , Triterpenos/efectos adversos , Triterpenos/uso terapéutico , Carga ViralRESUMEN
Under physiological conditions, mitochondrial morphology dynamically shifts between a punctuate appearance and tubular networks. However, little is known about upstream signal transduction pathways that regulate mitochondrial morphology. We show that mitochondrial fission is a very early and kinetically invariant event during neuronal cell death, which causally contributes to cytochrome c release and neuronal apoptosis. Using a small molecule CDK5 inhibitor, as well as a dominant-negative CDK5 mutant and RNAi knockdown experiments, we identified CDK5 as an upstream signalling kinase that regulates mitochondrial fission during apoptosis of neurons. Vice versa, our study shows that mitochondrial fission is a modulator contributing to CDK5-mediated neurotoxicity. Thereby, we provide a link that allows integration of CDK5 into established neuronal apoptosis pathways.
Asunto(s)
Apoptosis/fisiología , Quinasa 5 Dependiente de la Ciclina/metabolismo , Mitocondrias/enzimología , Neuronas/enzimología , Transducción de Señal/fisiología , Animales , Apoptosis/efectos de los fármacos , Células Cultivadas , Quinasa 5 Dependiente de la Ciclina/antagonistas & inhibidores , Quinasa 5 Dependiente de la Ciclina/genética , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Proteínas del Tejido Nervioso/metabolismo , Plásmidos , ARN Interferente Pequeño/genética , Ratas , Ratas Wistar , Transducción de Señal/efectos de los fármacos , Proteína bcl-X/metabolismo , Proteína bcl-X/farmacologíaRESUMEN
The microlymphocytotoxicity technique has been the accepted method for HLA class I typing since the early 1960s. However, it is often difficult to distinguish two related alleles expressed in an individual due to the cross-reactive nature of the alloantibodies used in this technique. This is especially evident at the HLA-B locus, whose more than 180 alleles fall into only 4 major interrelated cross-reactive antigen groups. To estimate the error rate in serologic typing due to the cross-reactive nature of sera, we used polymerase chain reaction with sequence-specific primers (PCR-SSP) amplification to retype 40 individuals who were previously typed as serologic HLA-B locus homozygotes. PCR-SSP revealed that 10 of these 40 individuals (25%) were actually heterozygous at their HLA-B loci. The HLA-B locus alleles of 9 of these 10 discrepant individuals were further analyzed by denaturing gradient gel electrophoresis followed by direct sequencing. The sequence analysis confirmed that all nine individuals were indeed HLA-B locus heterozygotes. This surprisingly high error rate in serologic definition of HLA-B molecules argues for the use of rapid DNA-based techniques in HLA class I typing, even in the setting of solid organ transplantation.
