Universität zu Köln

Diesner, Max and Neupert, Susanne (2018). Quantification of Biogenic Amines from Individual GFP-Labeled Drosophila Cells by MALDI-TOF Mass Spectrometry. Anal. Chem., 90 (13). S. 8035 - 8044. WASHINGTON: AMER CHEMICAL SOC. ISSN 1520-6882

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Abstract

Cell cell communication plays a crucial role in orchestrating and modulating neural circuits. To understand such interactions, it is vital to determine and quantify the involved messenger molecules such as neuropeptides and biogenic amines on the level of single cells. In this study, we used single-cell mass spectrometry (SCMS) to qualify and quantify octopamine (OA) and tyramine (TA) from isolated single cells from intact brains of the fruit fly Drosophila melanogaster. Our workflow involved targeted GFP-guided single-cell microdissection, on-plate chemical derivatization with 4-hydroxy-3-methoxycinnamaldehyde (CA) or 2,5-dimethyl-1H-pyrrole-3,4-dicarbaldehyde (DPD) for increasing ion stability and ion signal intensity, and isotopically marked internal standards for quantification by MALDI-TOF MS. We were able to determine a limit of detection for OA of 1 fmol/mu L, for TA of 2.5 fmol/mu L and a lower limit of quantification (LLOQ) of 10 fmol/mu L for both substances. SCMS of GFP-labeled somata from ventral midline neurons of the labial neuromere (VM1b) of the gnathal ganglion revealed an OA titer of 17.38 fmol/mu L and a TA titer (similar to 2.5 fmol/mu L) lower than the LLOQ, independent of sex. However, using a genetically altered driver line devoid of OA, Tssh(nM18)/Tdc2 > GFP, we confirmed TA in these cells. Furthermore, cold-anesthetization of flies caused a significant increase in OA content in VM1b somata. We compared OA titers of somata from two different OA/TA cell clusters to demonstrate the usefulness of targeted SCMS in advancing our understanding of OA/TA signaling in behavior and physiology. An influence on the detection of neuropeptides by our derivatized SCMS method could be excluded.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Diesner, Max UNSPECIFIED UNSPECIFIED UNSPECIFIED Neupert, Susanne UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-179974
DOI:	10.1021/acs.analchem.8b00961
Journal or Publication Title:	Anal. Chem.
Volume:	90
Number:	13
Page Range:	S. 8035 - 8044
Date:	2018
Publisher:	AMER CHEMICAL SOC
Place of Publication:	WASHINGTON
ISSN:	1520-6882
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language TISSUE CHEMICAL DERIVATIZATION; OCTOPAMINERGIC NEURONS; QUANTITATIVE-DETERMINATION; MATRIX; MELANOGASTER; METABOLITES; MOLECULES; PEPTIDES; BEHAVIOR; CIRCUIT Multiple languages Chemistry, Analytical Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/17997