Neuropeptidomics of the supraoptic rat nucleus.
The mammalian supraoptic nucleus (SON) is a neuroendocrine center in the brain regulating a variety of physiological functions. Within the SON, peptidergic magnocellular neurons that project to the neurohypophysis (posterior pituitary) are involved in controlling osmotic balance, lactation, and parturition, partly through secretion of signaling peptides such as oxytocin and vasopressin into the blood. An improved understanding of SON activity and function requires identification and characterization of the peptides used by the SON. Here, small-volume sample preparation approaches are optimized for neuropeptidomic studies of isolated SON samples ranging from entire nuclei down to single magnocellular neurons. Unlike most previous mammalian peptidome studies, tissues are not immediately heated or microwaved. SON samples are obtained from ex vivo brain slice preparations via tissue punch and the samples processed through sequential steps of peptide extraction. Analyses of the samples via liquid chromatography mass spectrometry and tandem mass spectrometry result in the identification of 85 peptides, including 20 unique peptides from known prohormones. As the sample size is further reduced, the depth of peptide coverage decreases; however, even from individually isolated magnocellular neuroendocrine cells, vasopressin and several other peptides are detected.
Bora
Adriana
A
Neuroscience Program, Department of Cell and Developmental Biology, Beckman Institute, and Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Annangudi
Suresh P
SP
Millet
Larry J
LJ
Rubakhin
Stanislav S
SS
Forbes
Andrew J
AJ
Kelleher
Neil L
NL
Gillette
Martha U
MU
Sweedler
Jonathan V
JV
D000818:Animals; D005260:Female; D008297:Male; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000737:chemistry; D009479:Neuropeptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D020318:Rats, Long-Evans; D013495:Supraoptic Nucleus / Q000737:chemistry* / Q000166:cytology
D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D020543:Proteome