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Master's Dissertation
DOI
https://doi.org/10.11606/D.17.2011.tde-17052011-164741
Document
Author
Full name
Suélen Merlo
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2011
Supervisor
Committee
Moreira, Jorge Eduardo (President)
Fernandes, Maria José da Silva
Leite, João Pereira
Title in Portuguese
Estudo dos grânulos de lipofucsina e das sinapses do córtex temporal durante o envelhecimento
Keywords in Portuguese
alfa-sinucleína
contatos sinápticos
córtex temporal humano e de ratos
grânulos de lipofucsina
microscopia confocal
microscopia de luz
microscopia eletrônica
sinapses
ubiquitina
vesículas sinápticas
western blots.
Abstract in Portuguese
Alterações morfológicas e funcionais ocorrem durante o envelhecimento, período da vida com maior incidência de doenças neurodegenerativas. No presente trabalho acompanhou-se a evolução dos grânulos de lipofucsina durante o envelhecimento para investigar alterações sinápticas, assim como proteínas associadas com doenças neurodegenerativas (alfa-sinucleína) e com o sistema ubiquitina-proteossoma em indivíduos de diferentes idades. No córtex temporal humano e de ratos determinou-se, nos diferentes grupos etários, seguindo a área, o número e as características dos grânulos de lipofucsina, o número de sinapses excitatórias, inibitórias e elétricas, os locais de contatos pós-sinápticos, o número de vesículas sinápticas por terminal e a expressão das proteínas alfa-sinucleína e ubiquitina. Amostras de córtex temporal humano de indivíduos com diferentes idades (20 - 28, 37 - 41 e 50 - 55 anos) foram coletadas de pacientes com epilepsia submetidos à lobectomia do lobo temporal. Amostras de ratos de 2, 6, 10 e 12 meses também foram coletadas. Foram utilizadas técnicas de microscopia de luz, eletrônica, confocal e western blots. Os dados obtidos de grânulos de lipofucsina são consistentes com outros estudos que observaram aumento dessa estrutura em mamíferos de maior idade. No entanto, os grânulos parecem crescer em volume, mas não em número, com aumento considerável da fração elétron lúcida (lipídica). Não houve diferença na expressão das proteínas alfa-sinucleína e ubiquitina entre os grupos das idades estudadas. A densidade sináptica foi similar entre os grupos experimentais, assim como o local de contato pós-sináptico. O aumento de vesículas elétron densas em sinapses inibitórias deve estar associado à demanda de neurotransmissores catecolaminérgicos. Estes resultados não expressam totalmente o processo de envelhecimento, pois as faixas etárias de humanos e ratos correspondem a uma idade ainda jovem. A coleta de material humano mais idoso foi impossibilitada pela faixa etária dos doentes submetidos à lobectomia. Os ratos do biotério da FMRP, não sobrevivem mais do que 12 meses em no nosso ambiente, incluindo manutenção dos animais isolados em racks.
Title in English
Study of lipofuscin granules and synapses in the temporal cortex during aging.
Keywords in English
alpha-synuclein
and electron microscopy
confocal
human and rat temporal cortex
light
lipofuscin granules
synapses
synaptic contacts
synaptic vesicles
ubiquitin
western blots.
Abstract in English
Morphological and functional changes occur during the aging, period of life with increased incidence of neurodegenerative diseases. Following the evolution of the lipofuscin granules along three periods of life in humans and rats, the present work investigated synaptic changes, as well as proteins associated with neurodegenerative diseases (alpha-synuclein) and the ubiquitin-proteossoma system in individuals of different ages. The objectives of the study were to analyze the temporal cortex of humans and rats: the number of excitatory, inhibitory, and electric synapses, the site of postsynaptic contacts, the number of synaptic vesicles per terminal, and the expression of the proteins alpha-synuclein and ubiquitin following the size and features of the lipofuscin granules. Samples of temporal cortex of human subjects with different ages (20-28, 37-41 and 50-55 years) were collected from patients with epilepsy who underwent temporal lobectomy. Samples from rats of 2, 6, 10 and 12 months were also collected. Light, confocal, and electron microscopy, and western blots techniques were used as procedures. The data obtained on lipofuscin granules were coincident with other studies that observed a higher area occupied by this structure in older mammals. However, the granules seem to grow in volume, but not in number, with considerable increase of the electron lucid fraction (lipidic). There was no difference in the alpha-synuclein and ubiquitin expressions between the experimental groups. The synaptic densities were similar between the groups, as well as the postsynaptic contacts. Increase of the electron dense vesicles in inhibitory synapses, appeared to be associated with the demand of catecholamines. These results do not express totally the aging process, because the range of age used in humans, and rats belong to a young age. The human samples from older ages was difficult because, in general, of the age of the patients submitted to lobotomy. The rats of the FMRP bioterium do not survived more that 12 months in our environment, even in controlled conditions.
 
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Dissertacao.pdf (6.18 Mbytes)
Publishing Date
2011-07-25
 
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