Abstract
Nuclear magnetic resonance (NMR)-based metabolomics is widely used in the research of metabolic conditions of complex biological systems under various conditions, and its use has been found in the field of male fertility. Here we describe the implementation of total and targeted NMR-based metabolomics in the research on Sertoli cell metabolism. Main principles and techniques of cell medium, cellular extracts, and intact cells are explained, as well as some classical experiments that can give complementary information on the Sertoli cell metabolism.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kovac JR, Pastuszak AW, Lamb DJ (2013) The use of genomics, proteomics, and metabolomics in identifying biomarkers of male infertility. Fertil Steril 99(4):998–1007. https://doi.org/10.1016/j.fertnstert.2013.01.111
Bieniek JM, Drabovich AP, Lo KC (2016) Seminal biomarkers for the evaluation of male infertility. Asian J Androl 18(3):426–433. https://doi.org/10.4103/1008-682x.175781
Jayaraman V, Ghosh S, Sengupta A, Srivastava S, Sonawat HM, Narayan PK (2014) Identification of biochemical differences between different forms of male infertility by nuclear magnetic resonance (NMR) spectroscopy. J Assist Reprod Genet 31(9):1195–1204. https://doi.org/10.1007/s10815-014-0282-4
Aaronson DS, Iman R, Walsh TJ, Kurhanewicz J, Turek PJ (2010) A novel application of 1H magnetic resonance spectroscopy: non-invasive identification of spermatogenesis in men with non-obstructive azoospermia. Hum Reprod 25(4):847–852. https://doi.org/10.1093/humrep/dep475
Dias TR, Alves MG, Rato L, Casal S, Silva BM, Oliveira PF (2016) White tea intake prevents prediabetes-induced metabolic dysfunctions in testis and epididymis preserving sperm quality. J Nutr Biochem 37:83–93. https://doi.org/10.1016/j.jnutbio.2016.07.018
Rocha CS, Martins AD, Rato L, Silva BM, Oliveira PF, Alves MG (2014) Melatonin alters the glycolytic profile of Sertoli cells: implications for male fertility. Mol Hum Reprod 20(11):1067–1076. https://doi.org/10.1093/molehr/gau080
Rato L, Alves MG, Socorro S, Carvalho RA, Cavaco JE, Oliveira PF (2012) Metabolic modulation induced by oestradiol and DHT in immature rat Sertoli cells cultured in vitro. Biosci Rep 32(1):61–69. https://doi.org/10.1042/bsr20110030
Kell DB, Brown M, Davey HM, Dunn WB, Spasic I, Oliver SG (2005) Metabolic footprinting and systems biology: the medium is the message. Nat Rev Microbiol 3(7):557–565. https://doi.org/10.1038/nrmicro1177
Boussouar F, Benahmed M (2004) Lactate and energy metabolism in male germ cells. Trends Endocrinol Metabol 15(7):345–350
Regueira M, Riera MF, Galardo MN, Pellizzari EH, Cigorraga SB, Meroni SB (2014) Activation of PPAR α and PPAR β/δ regulates Sertoli cell metabolism. Mol Cell Endocrinol 382(1):271–281
Gorga A, Rindone GM, Regueira M, Pellizzari EH, Camberos MC, Cigorraga SB, Riera MF, Galardo MN, Meroni SB (2017) PPARγ activation regulates lipid droplet formation and lactate production in rat Sertoli cells. Cell Tissue Res 369(3):611–624
Riera M (2002) Regulation of lactate production and glucose transport as well as of glucose transporter 1 and lactate dehydrogenase A mRNA levels by basic fibroblast growth factor in rat Sertoli cells. J Endocrinol 173(2):335–343
Galardo MN, Gorga A, Merlo JP, Regueira M, Pellizzari EH, Cigorraga SB, Riera MF, Meroni SB (2017) Participation of HIFs in the regulation of Sertoli cell lactate production. Biochimie 132:9–18
Fan TW-M (1996) Metabolite profiling by one- and two-dimensional NMR analysis of complex mixtures. Prog Nucl Magn Reson Spectrosc 28:161–219. https://doi.org/10.1016/0079-6565(95)01017-3
Barding GA Jr, Salditos R, Larive CK (2012) Quantitative NMR for bioanalysis and metabolomics. Anal Bioanal Chem 404(4):1165–1179. https://doi.org/10.1007/s00216-012-6188-z
Carrola J et al (2016) Metabolomics of silver nanoparticles toxicity in HaCaT cells: structure-activity relationships and role of ionic silver and oxidative stress. Nanotoxicology 10(8):1105–1117. https://doi.org/10.1080/17435390.2016.1177744
Duarte IF et al (2009) Analytical approaches toward successful human cell metabolome studies by NMR spectroscopy. Anal Chem 81(12):5023–5032. https://doi.org/10.1021/ac900545q
Ritter JB, Wahl AS, Freund S, Genzel Y, Reichl U (2010) Metabolic effects of influenza virus infection in cultured animal cells: intra- and extracellular metabolite profiling. BMC Syst Biol 4:61. https://doi.org/10.1186/1752-0509-4-61
Nowick JS, Khakshoor O, Hashemzadeh M, Brower JO (2003) DSA: a new internal standard for NMR studies in aqueous solution. Org Lett 5(19):3511–3513. https://doi.org/10.1021/ol035347w
Fan TW-M, Lane AN (2008) Structure-based profiling of metabolites and isotopomers by NMR. Prog Nucl Magn Reson Spectrosc 42:69–117. https://doi.org/10.1016/j.pnmrs.2007.03.002
Labarca C, Paigen K (1980) A simple, rapid, and sensitive DNA assay procedure. Anal Biochem 102(2):344–352
Acknowledgments
This work was supported by the Portuguese Foundation for Science and Technology (FCT) grant PTDC/BIM-MET/4712/2014 and by the Agencia Nacional de Promoción Cientíca y Tecnológica grants PICT 2014/945 and PICT2015/228 and by the Consejo Nacional de Investigaciones Cientícas y Técnicas (CONICET) grant PIP 2015/127.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Jarak, I. et al. (2018). Assessing Sertoli Cell Metabolic Activity. In: Alves, M., Oliveira, P. (eds) Sertoli Cells. Methods in Molecular Biology, vol 1748. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7698-0_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7698-0_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7697-3
Online ISBN: 978-1-4939-7698-0
eBook Packages: Springer Protocols