Efeito da melatonina sobre o perfil de miRNA de espermatozoides, regiões do cérebro e cerebelo de ratos Wistar
Data
2024-04-15
Autores
Nascimento, Mísia Helena da Silva Ferro 
Orientadores
Teixeira, Taiza Stumpp
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Introdução: A melatonina é sintetizada exclusivamente à noite, em diversos organismos. Nos mamíferos, a melatonina se origina nos pinealócitos presentes na glândula pineal. Dentre várias funções associadas a melatonina, a ação e influência do hormônio na espermatogênese é de particular interesse porque os gametas transferem não apenas informações genéticas, mas também epigenéticas para o embrião. Sabe-se que o espermatozoide é rico em miRNAs, que são transferidos para o embrião e podem configurar os mecanismos de herança epigenética paterna. Objetivo: investigar a influência da melatonina afeta a expressão de miRNA nos espermatozoides, regiões do cérebro e cerebelo de ratos. Metodologia: Esse estudo compreendeu três grupos experimentais: PINX, PINX MEL e controle. O primeiro consiste em animais que sofreram remoção da glândula pineal. O grupo PINX MEL foi composto por animais pinealectomizados que tiveram suplementação de melatonina. O grupo controle não sofreu manipulação. Os miRNAs foram extraídos dos espermatozoides, hipotálamo, córtex pré-frontal e cerebelo para análise de sua expressão por RT-qPCR. Foram analisados 8 miRNAs-alvos: miR-18a-5p, miR-34a-5p, miR-106b-5p, miR-132-5p, miR-195-5p, miR-219a-5p, miR-182-5p e Let-7g-5p. Foi realizada também análise de bioinformática buscando compreender as possíveis implicações biológicas das alterações da expressão dos miRNAs analisados. Resultados: Os animais pinealectomizados apresentaram redução da expressão de todos os miRNAs avaliados nos espermatozoides e de alguns desses miRNAs no cérebro e cerebelo. A administração da melatonina exógena levou à restauração da expressão desses miRNAs nos espermatozoides e também em algumas regiões do cérebro. A análise bioinformática indicou que todos os miRNAs analisados estão envolvidos em processos biológicos relacionados a neurogênese, sinapses e ao comportamento. Conclusão: O presente estudo sugere que a deficiência de melatonina afeta de forma significativa a expressão de miRNAs relacionados ao neurodesenvolvimento e à neurofisiologia em espermatozoides de ratos pinealectomizados. Estudos futuros são de grande valia a fim de esclarecer quais os significados biológicos dessas alterações para os animais pinealectomizados e se elas podem ter impacto para sua prole.
Introduction: Melatonin is synthesized exclusively at night, in several organisms. In mammals, melatonin originates in pinealocytes present in the pineal gland. Among several functions associated with melatonin, the action and influence of the hormone on spermatogenesis is of particular interest because gametes transfer not only genetic but also epigenetic information to the embryo. It is known that sperm is rich in miRNAs, which are transferred to the embryo and can configure paternal epigenetic inheritance mechanisms. Objective: to investigate the influence of melatonin affects the expression of miRNA in sperm, brain regions and cerebellum of rats. Methodology: This study comprised three experimental groups: PINX, PINX MEL and control. The first consists of animals that have had their pineal gland removed. The PINX MEL group was composed of pinealectomized animals that received melatonin supplementation. The control group was not manipulated. The miRNAs were extracted from sperm, hypothalamus, prefrontal cortex and cerebellum for analysis of their expression by RT-qPCR. 8 target miRNAs were analyzed: miR-18a-5p, miR-34a-5p, miR-106b-5p, miR-132-5p, miR-195-5p, miR-219a-5p, miR-182-5p and Let -7g-5p. Bioinformatics analysis was also carried out seeking to understand the possible biological implications of changes in the expression of the analyzed miRNAs. Results: Pinealectomized animals showed reduced expression of all miRNAs evaluated in sperm and some of these miRNAs in the brain and cerebellum. The administration of exogenous melatonin led to the restoration of the expression of these miRNAs in sperm and also in some regions of the brain. Bioinformatic analysis indicated that all analyzed miRNAs are involved in biological processes related to neurogenesis, synapses and behavior. Conclusion: The present study suggests that melatonin deficiency significantly affects the expression of miRNAs related to neurodevelopment and neurophysiology in sperm from pinealectomized rats. Future studies are of great value in order to clarify the biological meanings of these changes for pinealectomized animals and whether they can have an impact on their offspring.
Introduction: Melatonin is synthesized exclusively at night, in several organisms. In mammals, melatonin originates in pinealocytes present in the pineal gland. Among several functions associated with melatonin, the action and influence of the hormone on spermatogenesis is of particular interest because gametes transfer not only genetic but also epigenetic information to the embryo. It is known that sperm is rich in miRNAs, which are transferred to the embryo and can configure paternal epigenetic inheritance mechanisms. Objective: to investigate the influence of melatonin affects the expression of miRNA in sperm, brain regions and cerebellum of rats. Methodology: This study comprised three experimental groups: PINX, PINX MEL and control. The first consists of animals that have had their pineal gland removed. The PINX MEL group was composed of pinealectomized animals that received melatonin supplementation. The control group was not manipulated. The miRNAs were extracted from sperm, hypothalamus, prefrontal cortex and cerebellum for analysis of their expression by RT-qPCR. 8 target miRNAs were analyzed: miR-18a-5p, miR-34a-5p, miR-106b-5p, miR-132-5p, miR-195-5p, miR-219a-5p, miR-182-5p and Let -7g-5p. Bioinformatics analysis was also carried out seeking to understand the possible biological implications of changes in the expression of the analyzed miRNAs. Results: Pinealectomized animals showed reduced expression of all miRNAs evaluated in sperm and some of these miRNAs in the brain and cerebellum. The administration of exogenous melatonin led to the restoration of the expression of these miRNAs in sperm and also in some regions of the brain. Bioinformatic analysis indicated that all analyzed miRNAs are involved in biological processes related to neurogenesis, synapses and behavior. Conclusion: The present study suggests that melatonin deficiency significantly affects the expression of miRNAs related to neurodevelopment and neurophysiology in sperm from pinealectomized rats. Future studies are of great value in order to clarify the biological meanings of these changes for pinealectomized animals and whether they can have an impact on their offspring.
Descrição
Citação
NASCIMENTO, Mísia Helena da Silva Ferro. Efeito da melatonina sobre o perfil de miRNA de espermatozoides, regiões do cérebro e cerebelo de ratos Wistar. 2024. 95 f. Dissertação (Mestrado em Biologia Funcional e Estrutural) - Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, 2024.