Abstract

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.

Original languageEnglish (US)
Pages (from-to)4992-5003
Number of pages12
JournalJournal of Proteome Research
Volume7
Issue number11
DOIs
StatePublished - Nov 1 2008

Fingerprint

Supraoptic Nucleus
Rats
Peptides
Tandem Mass Spectrometry
Vasopressins
Neurons
Mass spectrometry
Brain
Tissue
Posterior Pituitary Gland
Neuroendocrine Cells
Liquid chromatography
Oxytocin
Lactation
Liquid Chromatography
Sample Size
Blood
Parturition

Keywords

  • ESI-MS/MS
  • Magnocellular neuron
  • MALDI-MS
  • Neuropeptides
  • Peptide processing
  • Peptidome
  • Prohormone
  • Sample preparation
  • Single cell
  • Supraoptic nuclei

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Bora, A., Annangudi, S. P., Millet, L. J., Rubakhin, S. S., Forbes, A. J., Kelleher, N. L., ... Sweedler, J. V. (2008). Neuropeptidomics of the supraoptic rat nucleus. Journal of Proteome Research, 7(11), 4992-5003. https://doi.org/10.1021/pr800394e

Neuropeptidomics of the supraoptic rat nucleus. / Bora, Adriana; Annangudi, Suresh P.; Millet, Larry J.; Rubakhin, Stanislav S.; Forbes, Andrew J.; Kelleher, Neil L.; Gillette, Martha U.; Sweedler, Jonathan V.

In: Journal of Proteome Research, Vol. 7, No. 11, 01.11.2008, p. 4992-5003.

Research output: Contribution to journalArticle

Bora, A, Annangudi, SP, Millet, LJ, Rubakhin, SS, Forbes, AJ, Kelleher, NL, Gillette, MU & Sweedler, JV 2008, 'Neuropeptidomics of the supraoptic rat nucleus', Journal of Proteome Research, vol. 7, no. 11, pp. 4992-5003. https://doi.org/10.1021/pr800394e
Bora A, Annangudi SP, Millet LJ, Rubakhin SS, Forbes AJ, Kelleher NL et al. Neuropeptidomics of the supraoptic rat nucleus. Journal of Proteome Research. 2008 Nov 1;7(11):4992-5003. https://doi.org/10.1021/pr800394e
Bora, Adriana ; Annangudi, Suresh P. ; Millet, Larry J. ; Rubakhin, Stanislav S. ; Forbes, Andrew J. ; Kelleher, Neil L. ; Gillette, Martha U. ; Sweedler, Jonathan V. / Neuropeptidomics of the supraoptic rat nucleus. In: Journal of Proteome Research. 2008 ; Vol. 7, No. 11. pp. 4992-5003.
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