Training in the state-of-the-art immunocytochemistry and its combination with non-radioactive in situ hybridization: expression and activation of melanin concentrating hormone after unremitting REM sleep deprivation in rats

• Grant number: 06/06289-9
• Support Opportunities: Scholarships abroad - Research
• Start date: February 26, 2007
• End date: May 25, 2007
• Field of knowledge: Biological Sciences - Morphology - Anatomy
• Principal Investigator: Luciane Valéria Sita
• Grantee: Luciane Valéria Sita
• Host Investigator: Gloria Ellyn Hoffman
• Host Institution: Pessoa Física
• Institution abroad: University of Maryland, Baltimore (UMB), United States

Abstract

This proposal represents a three month training experience for the application of immunocytochemistry (ICC, single and double labeling) and in situ hybridization (ISH, using biotinylated riboprobes). The focus of the immunocytochemical approaches will be to apply analysis of peptide expression for the melanin concentrating hormone (MCH) with that of the immediate early gene product Fos in rats that are either exposed to an unremitting rapid eye movement (REM) sleep deprivation paradigm for 10 days, or exposed and then allowed to recover briefly (2 hrs). The in situ hybridization approach will then use the same tissue to establish if there are changes in the level of MCH mRNA expression in the neurons and whether it varies among the various MCH populations. Our in situ hybridization methods enable quantitative assessment of expression while still retaining the ability to stain for Fos in the same sections. Thus, analyses of the distinct populations of neurons that are stimulated to express the immediate early genes can be compared with those that are quiescent at the time of euthanasia. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

KOBAN‚ M.; SITA‚ L.V.; LE‚ W.W.; HOFFMAN‚ G.E.. Sleep deprivation of rats: the hyperphagic response is real. Sleep, v. 31, n. 7, p. 927, 2008. (06/06289-9)