Collection of peptides released from single neurons with particle-embedded monolithic capillaries followed by detection with matrix-assisted laser desorption/ionization mass spectrometry

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Abstract

Characterization of the stimulated release of neuropeptides from brain slices and individual cultured neurons requires efficient collection of the releasate from relatively large volumes of physiological saline. Here, several collection approaches are optimized using particle-embedded monolithic capillaries (PEMCs) with poly(stearyl methacrylate-co-ethylene glycol dimethacrylate) monolith acting as a "glue". Two distinct extraction particles, with either pyrrolidone (PY) or ethylenediamine (EDA) as the functional group on polystyrene backbone, have been embedded into capillaries having an inner diameter of 250 μm. The capillaries act as collection devices for sampling neuropeptide release; the collection protocols are described, and the extraction efficiency of the probes are characterized. Specifically, the binding of angiotensin II from a peptide mixture onto the PY and EDA columns was 16 and 28 pmol, respectively, in a volume of 20 μL of saline. The peptides released from these columns have been characterized via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with low femtomole detection limits. When the PEMC columns were positioned in close proximity to individual neurons and 50 mM KCl was used as the secretagogue, peptides released from individual identified cultured neurons isolated from Aplysia californica were collected and characterized.

Original languageEnglish (US)
Pages (from-to)9557-9563
Number of pages7
JournalAnalytical chemistry
Volume83
Issue number24
DOIs
StatePublished - Dec 15 2011

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ethylenediamine
Neurons
Ionization
Mass spectrometry
Pyrrolidinones
Desorption
Neuropeptides
Peptides
Lasers
Glues
Polystyrenes
Angiotensin II
Functional groups
Brain
Sampling

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

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abstract = "Characterization of the stimulated release of neuropeptides from brain slices and individual cultured neurons requires efficient collection of the releasate from relatively large volumes of physiological saline. Here, several collection approaches are optimized using particle-embedded monolithic capillaries (PEMCs) with poly(stearyl methacrylate-co-ethylene glycol dimethacrylate) monolith acting as a {"}glue{"}. Two distinct extraction particles, with either pyrrolidone (PY) or ethylenediamine (EDA) as the functional group on polystyrene backbone, have been embedded into capillaries having an inner diameter of 250 μm. The capillaries act as collection devices for sampling neuropeptide release; the collection protocols are described, and the extraction efficiency of the probes are characterized. Specifically, the binding of angiotensin II from a peptide mixture onto the PY and EDA columns was 16 and 28 pmol, respectively, in a volume of 20 μL of saline. The peptides released from these columns have been characterized via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with low femtomole detection limits. When the PEMC columns were positioned in close proximity to individual neurons and 50 mM KCl was used as the secretagogue, peptides released from individual identified cultured neurons isolated from Aplysia californica were collected and characterized.",
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