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Int. J. Mol. Sci. 2019, 20(2), 314; https://doi.org/10.3390/ijms20020314

Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS

1
American Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH 44195, USA
2
Universidade da Beira Interior, 6201-001 Covilhã, Portugal
3
Department of Microscopy, Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar and Unit for Multidisciplinary Research in Biomedicine, University of Porto, 4050-313 Porto, Portugal
4
Redox Biology Laboratory, Department of Zoology, School of Life Sciences, Ravenshaw University, Cuttack 753003, India
*
Author to whom correspondence should be addressed.
Received: 24 December 2018 / Revised: 8 January 2019 / Accepted: 10 January 2019 / Published: 14 January 2019
(This article belongs to the Section Molecular Biology)
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Abstract

In sperm proteomic experiments round cells and leukocyte proteins are profiled along with sperm proteome. The influence of round cell and leukocyte proteins on the sperm proteome has not been investigated. The objective of this study was to identify if the proteins from round cells, including leukocytes, interfere with the proteomic analysis of spermatozoa in frozen semen samples. Proteomic profiling of sperm was performed using liquid chromatography-tandem mass spectrometry in four groups: Group 1 contained neat semen with round cells and leukocytes ≥ 1 × 106/mL, group 2 contained neat semen with round cells ≥ 1 × 106/mL that was processed by 65% density gradient to remove the round cells and leukocytes, group 3 contained neat semen with round cells < 1 × 106/mL, and group 4 contained neat semen with round cells < 1 × 106/mL that was processed by 65% density gradient to remove the round cells. Pure leukocyte culture was used as control group. A total of 1638, 1393, 1755, and 1404 proteins were identified in groups 1, 2, 3, and 4, respectively. Comparative analysis of group 1 vs. 3 revealed 26 (1.18%) differentially expressed proteins (DEPs). On the other hand, only 6 (0.31%) DEPs were observed with group 2 vs. 4. Expression of these DEPs were either absent or very low in the control group. The results of our proteomics analysis failed to show any influence of non-spermatogenic round cell proteins on sperm proteome identification. These results validate the use of neat semen samples for sperm proteomic studies. View Full-Text
Keywords: sperm; leukocytes; round cells; non-spermatogenic cells; frozen semen; proteomics sperm; leukocytes; round cells; non-spermatogenic cells; frozen semen; proteomics
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Panner Selvam, M.K.; Agarwal, A.; Dias, T.R.; Martins, A.D.; Samanta, L. Presence of Round Cells Proteins do not Interfere with Identification of Human Sperm Proteins from Frozen Semen Samples by LC-MS/MS. Int. J. Mol. Sci. 2019, 20, 314.

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