Identification and Relative Quantification of Neuropeptides from the Endocrine Tissues
Endocrine tissues like the pituitary, hypothalamus and islets of Langerhans are rich in bioactive peptides. These are used for intercellular signalling and are involved in regulation of almost all physiological processes. Peptidomics is the comprehensive analysis of peptides in tissues, fluids and cells. Peptidomics applied to (neuro-)endocrine tissues aims therefore to identify as many bioactive peptides as possible. Peptidomics of (neuro-)endocrine tissues requires an integrated approach that consists of careful sample handling, peptide separation techniques, mass spectrometry and bioinformatics. Here we describe the methods for isolation and dissection of endocrine tissues, the extraction of bioactive peptides and further sample handling and identification of peptides by mass spectrometry and hyphenated techniques. We also present a straightforward method for the comparison of relative levels of bioactive peptides in these endocrine tissues under varying physiological conditions. The latter helps to elucidate functions of the bioactive peptides.
Key wordsNeuropeptide Mus musculus peptidomics pituitary hypothalamus islets of Langerhans endocrine pancreas MALDI-TOF/TOF MS Q-TOF MS two-dimensional HPLC nanoLC
K. Boonen and B. Landuyt are supported by grants of Institute for the Promotion of Innovation through Science and Technology (I.W.T.)-Flanders. S. J. Husson is a postdoctoral fellow of the Research Foundation Flanders (F.W.O.-Vlaanderen). The authors also acknowledge the Interfacultary Centre for Proteomics and Metabolomics “Prometa”, K.U. Leuven.
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