Dados do Trabalho

Título

Neuronal density may have a relationship with focal cortical dysplasia refractory epilepsy

Introdução

The cortical dysplasia is one of the most common forms of malformations of the cortex development, being highly associated with refractory epilepsy. The focal cortical dysplasia (FCD) is formed by heterogeneous groups of cortical lesions characterized mostly in abnormal cortical architecture or cytologic abnormalities. As a form of quantification of these abnormalities, the neuronal density of dysplastic cortex samples can be measured and used as data.

Objetivo

Investigate possible changes in the neuronal density in samples of cerebral cortex with focal cortical dysplasia.

Método

For this study, the dysplastic brain tissue of 7 patients with focal cortical dysplasia (FCD) were collected via surgical resection for cell counting. The fragment of samples were weighted and mechanically dissociated by maceration in a solution of sodium citrate 40 mM and Triton X-100 1%. The homogenized solution was centrifuged for 10 minutes at 4000 x g and the nuclei were collected from the pellet formed in the tube. Pelletized nuclei were suspended in a solution of Phosphate Buffered Saline (PBS) with 1% of 4′,6-diamidino-2-phenylindole (DAPI), a fluorescent dye specific for DNA. The final volume was adjusted of the suspension to create a fitting nuclei density for the hemocytometer and then the nuclei were counted using a fluorescence microscope with a 400x zoom. To a fraction of this suspension, it will be added the NeuN antibody for neurons identification and the differentiated count between total cells and neurons present in the sample will be performed.

Resultados

The nuclei were identified in the hemocytometer using the microscope and were able to be counted. Now, the protocols are being adapted for better results.

Conclusão

Considering the importance of studies about FCD and its effects on cortical architecture and cytology, the neuronal density estimation may have a relationship with refractory epilepsy in these patients.

Palavras-chave

Focal cortical dysplasia, hemocytometer, neuronal density

Área

Psicoterapia e Neurociências