Proteomic profiling of the rat hypothalamus

Proteome Science

Eliane B Ribeiro²  Dulce E Casarini¹  José C Rosa⁵  Lila M Oyama⁶  Cláudia M O Nascimento⁶  Debora Estadella⁶  Mariana L Lima⁴  Maísa M Sakata¹  Juliana D Perez¹  Maria C C Andrade¹  Mônica M Telles³  Kelse T Albuquerque⁶  Regina L H Watanabe⁶  Amanda P Pedroso⁶ 

[1] Department of Medicine, Division of Nephrology, Universidade Federal de São Paulo, UNIFESP, São Paulo, SP, Brazil;Departamento de Fisiologia, Universidade Federal de São Paulo – Escola Paulista de Medicina, Rua Botucatu 862 2° andar, Vila Clementino, 04023-062, São Paulo, SP, Brazil;Department of Biological Sciences, Universidade Federal de São Paulo, UNIFESP, São Paulo, SP, Brazil;Department of Microbiology, Immunology and Parasitology, Universidade Federal de São Paulo, UNIFESP, São Paulo, SP, Brazil;Centro de Química de Proteínas, FMRP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil;Department of Physiology, Division of Nutrition Physiology, Universidade Federal de São Paulo, UNIFESP, São Paulo, SP, Brazil

关键词: Rat;    Obesity;    Nutrition;    Mass spectrometry;    2-D electrophoresis;    Proteome;    Hypothalamus;   

Others  :  817309
DOI  :  10.1186/1477-5956-10-26

 received in 2011-12-02, accepted in 2012-04-20,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Pedroso et al.; licensee BioMed Central Ltd.

【 图 表 】

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