Abstract
Freshwater planarian species S. mediterranea is an emerging stem cell model because of its capability of regenerating large portions of missing body parts. It is one of the best model systems available to address the basic biological mechanisms in the regeneration processes. Absolute quantification of metabolites from planarians is imperative to understand their role in the regeneration processes. Here we describe a stable isotope dilution ultrahigh performance liquid chromatography/mass spectrometry/selected reaction monitoring (UHPLC-MS/SRM) assay for a sensitive and quantitative assessment of neurotransmitters (NTs) in planaria. We used this method for the simultaneous quantification of 16 NTs from both intact and regenerating planarians.
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References
Newmark PA, Alvarado AS (2002) Not your father’s planarian: a classic model enters the era of functional genomics. Nat Rev Genet 3:210–219. https://doi.org/10.1038/nrg759
Agata K, Tanaka T, Kobayashi C et al (2003) Intercalary regeneration in planarians. Dev Dyn 226:308–316. https://doi.org/10.1002/dvdy.10249
Cebrià F, Nakazawa M, Mineta K et al (2002) Dissecting planarian central nervous system regeneration by the expression of neural-specific genes. Develop Growth Differ 44:135–146. https://doi.org/10.1046/j.1440-169x.2002.00629.x
Cebrià F, Newmark PA (2007) Morphogenesis defects are associated with abnormal nervous system regeneration following roboA RNAi in planarians. Development 134:833–837. https://doi.org/10.1242/dev.02794
Agata K, Soejima Y, Kato K et al (1998) Structure of the planarian central nervous system (CNS) revealed by neuronal cell markers. Zool Sci 15:433–440. https://doi.org/10.2108/zsj.15.433
Reuter M, Gustafsson MKS (1995) The flatworm nervous system: pattern and phylogeny. Experientia Suppl 72:25–59. https://doi.org/10.1007/978-3-0348-9219-3_3
Collins JJ III, Hou X, Romanova EV et al (2010) Genome-wide analyses reveal a role for peptide hormones in planarian germline development. PLoS Biol 8:e1000509. https://doi.org/10.1371/journal.pbio.1000509
Ribeiro P, El-Shehabi F, Patocka N (2005) Classical transmitters and their receptors in flatworms. Parasitology 131:S19–S40. https://doi.org/10.1017/S0031182005008565
Kannan R, Dasaradhi P (2013) Comprehensive analysis of neurotransmitters from regenerating planarian extract using an ultrahigh-performance liquid chromatography/mass spectrometry/selected reaction monitoring method. Rapid Commun Mass Spectrom. 27:2439–2452. https://doi.org/10.1002/rcm.6706
Acknowledgments
We thank the Department of Biotechnology (DBT), Government of India for financial assistance to establish the metabolomics facility at Centre for Cellular and Molecular Platforms (C-CAMP), NCBS campus, Bangalore.
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Rangiah, K., Palakodeti, D. (2018). Quantification of Neurotransmitters from Intact and Regenerating Planarians Using UHPLC-MS/SRM Method. In: Rink, J. (eds) Planarian Regeneration. Methods in Molecular Biology, vol 1774. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7802-1_25
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DOI: https://doi.org/10.1007/978-1-4939-7802-1_25
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Publisher Name: Humana Press, New York, NY
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