Title: Metabolic Noise and Distinct Subpopulations Observed by Single Cell LAESI Mass Spectrometry of Plant Cells in situ

Abstract

Phenotypic variations and stochastic expression of transcripts, proteins, and metabolites in biological tissues lead to cellular heterogeneity. As a result, distinct cellular subpopulations emerge. They are characterized by different metabolite expression levels and by associated metabolic noise distributions. To capture these biological variations unperturbed, highly sensitive in situ analytical techniques are needed that can sample tissue embedded single cells with minimum sample preparation. Optical fiber-based laser ablation electrospray ionization mass spectrometry (f-LAESI-MS) is a promising tool for metabolic profiling of single cells. Integration of this MS-based platform with fluorescence and brightfield microscopy provides the ability to target single cells of specific type and allows for the selection of rare cells, e.g., excretory idioblasts. Analysis of individual Egeria densa leaf blade cells (n = 103) by f LAESI MS revealed significant differences between the prespecified subpopulations of epidermal cells (n = 97) and excretory idioblasts (n = 6) that otherwise would have been masked by the population average. Primary metabolites, e.g., malate, aspartate, and ascorbate, as well as several glucosides were detected in higher abundance in the epidermal cells. The idioblasts contained lipids, e.g., PG(16:0/18:2), and triterpene saponins, e.g., medicoside I and azukisaponin I, and their isomers. Metabolic noise for themore » epidermal cells were compared to results for soybean (Glycine max) root nodule cells (n = 60) infected by rhizobia (Bradyrhizobium japonicum). Whereas some primary metabolites showed lower noise in the latter, both cell types exhibited higher noise for secondary metabolites. Post-hoc grouping of epidermal and root nodule cells, based on the abundance distributions for certain metabolites (e.g., malate), enabled the discovery of cellular subpopulations characterized by different mean abundance values, and the magnitudes of the corresponding metabolic noise. Comparison of prespecified populations from epidermal cells of the closely related E. densa (n = 20) and Elodea canadensis (n = 20) revealed significant differences, e. g., higher sugar content in the former and higher levels of ascorbate in the latter, and the presence of species-specific metabolites. These results demonstrate that the f-LAESI-MS single cell analysis platform has the potential to explore cellular heterogeneity and metabolic noise for hundreds of tissue-embedded cells.« less

Authors:

Stopka, Sylwia A. ^[1]; Khattar, Rikkita ^[1]; Agtuca, Beverly J. ^[2]; Anderton, Christopher R. ^[3]; Pasa Tolic, Ljiljana ^[3]; Stacey, Gary ^[2]; Vertes, Akos ^[1]

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+ Show Author Affiliations

1. George Washington Univ., Washington, DC (United States)
2. Univ. of Missouri, Columbia, MO (United States)
3. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

Publication Date:

2018-11-15

Research Org.:

Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

Sponsoring Org.:

USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

OSTI Identifier:

1482272

Alternate Identifier(s):

OSTI ID: 1507727

Report Number(s):

PNNL-SA-137441
Journal ID: ISSN 1664-462X

Grant/Contract Number:  

AC05-76RL01830; SC0013978

Resource Type:

Published Article

Journal Name:

Frontiers in Plant Science

Additional Journal Information:

Journal Volume: 9; Journal ID: ISSN 1664-462X

Publisher:

Frontiers Research Foundation

Country of Publication:

United States

Language:

English

Subject:

54 ENVIRONMENTAL SCIENCES; 59 BASIC BIOLOGICAL SCIENCES