RESUMO
Function of dystrophin Dp71 isoforms is unknown but seems related to neurite outgrowth and synapse formation. To evaluate Dp71 role in myoneural synapses, we established a coculture model using PC12 cells and L6 myotubes and analyzed expression and localization of Dp71 and related proteins, utrophin and beta-dystroglycan, in PC12 cells. Confocal microscopy showed Dp71d isoform in PC12 nuclei, golgi-complex-like and endoplasmic reticulum-like structures, whereas Dp71ab concentrates at neurite tips and cytoplasm, colocalizing with beta-dystroglycan, utrophin, synaptophysin and acetylcholine receptors. Evidences suggest that Dp71ab isoform, unlike Dp71d, may take part in neurite-related processes. This is the first work on Dp and members of Dp-associated protein complex roles in a cell-line based coculturing system, which may be useful in determining Dp71 isoforms associations.
Assuntos
Distroglicanas/metabolismo , Distrofina/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Junção Neuromuscular/embriologia , Neurônios/metabolismo , Utrofina/metabolismo , Animais , Diferenciação Celular/fisiologia , Técnicas de Cocultura , Distroglicanas/genética , Distrofina/genética , Microscopia Confocal , Fibras Musculares Esqueléticas/ultraestrutura , Músculo Esquelético/embriologia , Músculo Esquelético/inervação , Músculo Esquelético/metabolismo , Neuritos/metabolismo , Neuritos/ultraestrutura , Junção Neuromuscular/citologia , Junção Neuromuscular/metabolismo , Neurônios/ultraestrutura , Organelas/metabolismo , Organelas/ultraestrutura , Células PC12 , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Ratos , Receptores Colinérgicos/metabolismo , Membranas Sinápticas/metabolismo , Membranas Sinápticas/ultraestrutura , Sinaptofisina/metabolismo , Utrofina/genéticaRESUMO
Three highly informative markers genetically linked to Huntington's Disease (HD) were used for diagnosis of HD in Mexican patients, two polymorphic HindIII sites located at D4S10 locus and one VNTR marker at D4S111 locus (VNTR-111). Forty chromosomes from healthy sybjects were tested in order to evaluate the informativeness of the probes. The RFLP HindIII 1 and 2 and the VNTR-111 probes showed a heterozygosity of 0 percent, 45 percent, and 60 percent, respectively. Five families were analyzed, of these, only in two the markers used were informative. In one of them, six membrers showed a decreased risk of inheritance of the mutant gene for Huntington's Didease with 95 percent accuracy
Assuntos
Humanos , DNA , Doenças Genéticas Inatas/terapia , Genética Médica/métodos , Genética Populacional , Doença de Huntington/diagnóstico , Marcadores Genéticos/fisiologia , México , Biologia Molecular , Reação em Cadeia da Polimerase , Fatores de RiscoRESUMO
The frequency of allels, intragenic (intron 18) and extrangenic (DXS52) Bcl I RFLPs was investigated in a sample of the Mexican population. Altogether 33 X chromosomes at R8c locus and 30 at DXS52 locus were studied. The allele frequencies found at the F8c locus were similar to those reported in the majority of other populations. The observed heterozygosity for the intragenic extragenic markers were 0.57 and 0.64, respectively. By using these two RFLps 15 famales at risk in five independent families with hemophilia A were investigated; ten for them could be identified and five excluded as carries
