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Master's Dissertation
DOI
10.11606/D.42.2013.tde-20092013-112432
Document
Author
Full name
Jessica Andrade da Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Cipolla Neto, Jose (President)
Castrucci, Ana Maria de Lauro
Lima, Fabio Bessa
Title in Portuguese
Papel da temporização noradrenérgica na regulação da síntese de melatonina pela glândula pineal em cultura: características funcionais e mecanismos de ação.
Keywords in Portuguese
Cronobiologia
Genes relógio
Glândula Pineal
Melatonina
Noradrenalina
Abstract in Portuguese
A glândula pineal de mamíferos não é uma estrutura oscilatória autônoma, exigindo a liberação de noradrenalina (Nor) na fase escura para que a melatonina seja circadianamente produzida. Na cultura padrão de glândula pineal, o órgão não expressa ritmicidade funcional e para mimetizar o padrão fisiológico de liberação de Nor, desenvolvemos a cultura temporizada com Nor. Logo, o objetivo desse trabalho foi investigar a manutenção do ritmo de expressão dos genes relógio pela cultura temporizada, e qual a via noradrenérgica envolvida. Para os estudos in vitro, realizaram-se culturas dos grupos: controle (sem Nor), agudo (cultura padrão) e temporizado (12h com Nor/12h sem Nor). Além disso, à cultura temporizada se adicionou Prasozin e/ou Propranolol. Analisou-se expressão dos genes relógio, atividade da enzima AANAT e conteúdo de melatonina no meio de cultura. No grupo temporizado, observou-se a manutenção da ritmicidade dos genes analisados, diferente do observado nos grupos controle, agudo e temporizado tratado com bloqueadores, além do aumento da atividade enzimática da AANAT e aumento do conteúdo de melatonina. Em suma, a cultura temporizada com Nor se mostra importante para evitar a arritmicidade encontrada na cultura padrão de glândula pineal.
Title in English
Role of norepinephrine synchronization on melatonin synthesis regulation of pineal gland culture: function and action mechanisms.
Keywords in English
Chronobiology
Clock genes
Melatonin
Norepinephrine
Pineal gland
Abstract in English
The mammals pineal gland is not an autonomous oscillator, the circadian melatonin synthesis requires the release of norepinephrine (NE) on the dark phase. In standard pineal gland culture, the glands do not express any functional rhythmicity. To mimic the physiological pattern of NE release in the pineal gland culture, we developed a synchronized culture with NE. We aimed to investigate the maintenance of circadian clock genes expression within rat pineal gland under acute and synchronized culture and the noradrenergic pathway involved. In in vitro experiments, culture glands were under: control (without NE), acute (standard culture) and synchronized (12h with NE/12h without NE) conditions. Furthermore, in the synchronized group were added Prasozin and/or Propranolol. We investigated clock genes expression, AANAT activity and melatonin content. The synchronized culture was able to maintain the rhythmic clock genes expression, which didn´t occur in control, acute and synchronized treated with blockers groups, and was able to improve AANAT activity and melatonin synthesis. In conclusion, synchronized culture method showed as a useful approach to avoid disruption of rhythmic variations found in the standard culture.
 
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Publishing Date
2013-12-05
 
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