Proteomic profiling of the rat hypothalamus

Proteomic profiling of the rat hypothalamus

Author Pedroso, Amanda P. Autor UNIFESP Google Scholar
Watanabe, Regina L. H. Autor UNIFESP Google Scholar
Albuquerque, Kelse T. Autor UNIFESP Google Scholar
Telles, Monica M. Autor UNIFESP Google Scholar
Andrade, Maria C. C. Autor UNIFESP Google Scholar
Perez, Juliana D. Autor UNIFESP Google Scholar
Sakata, Maisa M. Autor UNIFESP Google Scholar
Lima, Mariana L. Autor UNIFESP Google Scholar
Estadella, Debora Autor UNIFESP Google Scholar
Nascimento, Claudia Maria da Penha Oller do Autor UNIFESP Google Scholar
Oyama, Lila Missae Autor UNIFESP Google Scholar
Rosa, Jose C. Autor UNIFESP Google Scholar
Casarini, Dulce E. Autor UNIFESP Google Scholar
Ribeiro, Eliane B. Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Universidade de São Paulo (USP)
Abstract Background: the hypothalamus plays a pivotal role in numerous mechanisms highly relevant to the maintenance of body homeostasis, such as the control of food intake and energy expenditure. Impairment of these mechanisms has been associated with the metabolic disturbances involved in the pathogenesis of obesity. Since rodent species constitute important models for metabolism studies and the rat hypothalamus is poorly characterized by proteomic strategies, we performed experiments aimed at constructing a two-dimensional gel electrophoresis (2-DE) profile of rat hypothalamus proteins.Results: As a first step, we established the best conditions for tissue collection and protein extraction, quantification and separation. the extraction buffer composition selected for proteome characterization of rat hypothalamus was urea 7 M, thiourea 2 M, CHAPS 4%, Triton X-100 0.5%, followed by a precipitation step with chloroform/methanol. Two-dimensional (2-D) gels of hypothalamic extracts from four-month-old rats were analyzed; the protein spots were digested and identified by using tandem mass spectrometry and database query using the protein search engine MASCOT. Eighty-six hypothalamic proteins were identified, the majority of which were classified as participating in metabolic processes, consistent with the finding of a large number of proteins with catalytic activity. Genes encoding proteins identified in this study have been related to obesity development.Conclusion: the present results indicate that the 2-DE technique will be useful for nutritional studies focusing on hypothalamic proteins. the data presented herein will serve as a reference database for studies testing the effects of dietary manipulations on hypothalamic proteome. We trust that these experiments will lead to important knowledge on protein targets of nutritional variables potentially able to affect the complex central nervous system control of energy homeostasis.
Keywords Hypothalamus
2-D electrophoresis
Mass spectrometry
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Date 2012-04-20
Published in Proteome Science. London: Biomed Central Ltd, v. 10, 15 p., 2012.
ISSN 1477-5956 (Sherpa/Romeo, impact factor)
Publisher Biomed Central Ltd
Extent 15
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000308914400001

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