Use of sensitivity-enhanced nuclear magnetic resonance spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe in identifying human sperm intracellular metabolites
Aswathi Cheredath A , Shubhashree Uppangala
B , Ameya Jijo A , R. Vani Lakshmi C , G. A. Nagana Gowda D , Guruprasad Kalthur E and Satish Kumar Adiga A *
A Centre of Excellence in Clinical Embryology, Department of Reproductive Science, Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal 576 104, India.
B Division of Reproductive Genetics, Department of Reproductive Science, Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal 576 104, India.
C Department of Data Science, Prasanna School of Public Health, Manipal Academy of Higher Education, Manipal 576 104, India.
D Northwest Metabolomics Research Centre, Anaesthesiology and Pain Medicine, University of Washington, Seattle, WA, USA.
E Division of Reproductive Biology, Department of Reproductive Science, Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal 576 104, India.
Reproduction, Fertility and Development 35(14) 661-668 https://doi.org/10.1071/RD22246
Published online: 30 August 2023
Abstract
The clinical value of human sperm metabolites has not been established due to the technical complexity in detecting these metabolites when sperm numbers are low.
To detect endogenous intracellular metabolites in fresh and post-thaw human spermatozoa using 800 MHz nuclear magnetic resonance (NMR) spectroscopy equipped with a 1.7-mm cryo-probe.
Processed spermatozoa from 25 normozoospermic ejaculates were subjected to extraction of intracellular metabolites and then profiled by sensitivity-enhanced NMR spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe. In parallel, some of the processed sperm fractions were subjected to freeze-thawing and were then analysed for intracellular metabolites.
Twenty-three metabolites were profiled from only 1.25 million sperm cells. Comparison of the metabolomic signature of pre-freeze and post-thaw sperm cells did not show significant changes in the levels of metabolites.
Sensitivity-enhanced NMR spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe is a potential tool for identifying intracellular metabolites when sperm number is low.
Use of sensitivity-enhanced NMR spectroscopy opens up the opportunity to test for endogenous metabolites in samples with a limited number of spermatozoa, to understand the patho-physiology of infertility.
Keywords: cryopreservation, human spermatozoa, intracellular metabolites, male fertility, metabolomics NMR spectroscopy, semen, sperm function.
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