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. 2019;8(1):A0072.
doi: 10.5702/massspectrometry.A0072. Epub 2019 Feb 14.

Measurement of the Spatial Distribution of S1P in Small Quantities of Tissues: Development and Application of a Highly Sensitive LC-MS/MS Method Combined with Laser Microdissection

Jiao Wang  1 , Kuniyuki Kano  1   2 , Daisuke Saigusa  3   4   2 , Junken Aoki  1   2
Affiliations

Measurement of the Spatial Distribution of S1P in Small Quantities of Tissues: Development and Application of a Highly Sensitive LC-MS/MS Method Combined with Laser Microdissection

Jiao Wang et al. Mass Spectrom (Tokyo). 2019.

Abstract

Sphingosine-1-phosphate (S1P) acts as an extracellular signaling molecule with diverse biological functions. Tissues appear to have an S1P gradient, which is functionally relevant in the biological significance of S1P, although its existence has not been measured directly. Here, we report a highly sensitive method to determine the distribution of S1P, using a column-switching LC-MS/MS system combined with laser microdissection (LMD). Column switching using narrow core Capcell Pak C18 analytical and trap columns with 0.3 mm inner diameter improved the performance of the LC-MS/MS system. The calibration curve of S1P showed good linearity (r>0.999) over the range of 0.05-10 nM (1-200 fmol/injection). The accuracy of the method was confirmed by measuring S1P-spiked laser microdissected mice tissue sections. To evaluate our S1P analytical method, we quantified S1P extracted from micro-dissected mouse brain and spleen. These results show that this method can measure low S1P concentrations and determine S1P distribution in tissue microenvironments.

Keywords: column-switching LC-MS/MS; laser microdissection; sphingosine-1-phosphate.

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Figures

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Fig. 1. Performance of optimized LC-MS/MS systems for S1P quantification.
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Fig. 2. Comparison of the calculated and spiked S1P concentrations in different tissues.
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Fig. 3. S1P distribution in brain tissue sections.
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Fig. 4. S1P distribution in spleen tissue sections.
See this image and copyright information in PMC

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References

    1. M. Maceyka, K. B. Harikumar, S. Milstien, S. Spiegel. Sphingosine-1-phosphate signaling and its role in disease. Trends Cell Biol. 22: 50–60, 2012. - PMC - PubMed
    1. V. A. Blaho, T. Hla. An update on the biology of sphingosine 1-phosphate receptors. J. Lipid Res. 55: 1596–1608, 2014. - PMC - PubMed
    1. M. Książek, M. Chacinska, A. Chabowski, M. Baranowski. Sources, metabolism, and regulation of circulating sphingosine-1-phosphate. J. Lipid Res. 56: 1271–1281, 2015. - PMC - PubMed
    1. Y. Hisano, N. Kobayashi, A. Yamaguchi, T. Nishi. Mouse SPNS2 functions as a sphingosine-1-phosphate transporter in vascular endothelial cells. PLoS ONE 7: e38941, 2012. - PMC - PubMed
    1. S. R. Schwab, J. P. Pereira, M. Matloubian, Y. Xu, Y. Huang, J. G. Cyster. Lymphocyte sequestration through S1P lyase inhibition and disruption of S1P gradients. Science 309: 1735–1739, 2005. - PubMed