Neuropeptide expression and processing as revealed by direct matrix-assisted laser desorption ionization mass spectrometry of single neurons.
Neuropeptides were directly detected in single identified neurons and the neurohemal area of peptidergic (neuroendocrine) systems in the Lymnaea brain by using matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The samples were placed in matrix solution and ruptured to allow mixing of cell contents with the matrix solution. After formation of matrix crystals, the analytes were analyzed by MALDI-MS. It was surprising that clean mass spectra were produced, displaying extreme sensitivity of detection. In one of the neuroendocrine systems studied, we could demonstrate for the first time, by comparing the peptide patterns of soma and of neurohemal axon terminals, that processing of the complex prohormone expressed in this system occurs entirely in the soma. In the other system studied, novel peptides could be detected in addition to peptides previously identified by conventional molecular biological and peptide chemical methods. Thus, complex peptide processing and expression patterns could be predicted that were not detected in earlier studies using conventional methods. As the first MALDI-MS study of direct peptide fingerprinting in the single neuron, these experiments demonstrate that MALDI-MS forms a new and valuable approach to the study of the synthesis and expression of bioactive peptides, with potential application to single-cell studies in vertebrates, including humans.
Jiménez
C R
CR
Graduate School Neurosciences Amsterdam, Research Institute Neurosciences, Vrije Universiteit, Faculty of Biology, The Netherlands.
van Veelen
P A
PA
Li
K W
KW
Wildering
W C
WC
Geraerts
W P
WP
Tjaden
U R
UR
van der Greef
J
J
D000818:Animals; D001369:Axons / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D005260:Female; D007447:Invertebrate Hormones / Q000096:biosynthesis*; D007834:Lasers; D008195:Lymnaea; D013058:Mass Spectrometry; D009411:Nerve Endings / Q000378:metabolism; D009420:Nervous System / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D009479:Neuropeptides / Q000096:biosynthesis*; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational
D007447:Invertebrate Hormones; D009479:Neuropeptides; D011498:Protein Precursors; C042422:ovulation hormone, Lymnaea; D002118:Calcium